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Sample records for carcinomas methylation status

  1. Helicobacter pylori and EBV in gastric carcinomas:Methylation status and microsatellite instability

    Institute of Scientific and Technical Information of China (English)

    Adriana; Camargo; Ferrasi; Nídia; Alice; Pinheiro; Silvia; Helena; Barem; Rabenhorst; Otávia; Luisa; Caballero; Maria; Aparecida; Marchesan; Rodrigues; Fabrício; de; Carvalho; Celso; Vieira; de; Souza; Leite; Marcia; Valéria; Pitombeira; Ferreira; Marcos; Aurélio; Pessoa; Barros; Maria; Inês; de; Moura; Campos; Pardini

    2010-01-01

    AIM:To verify the methylation status of CDH1, DAPK, COX2, hMLH1 and CDKN2A genes and to evaluate their association with Helicobacter pylori (H. pylori)-cagA+ and Epstein Barr virus (EBV) infections in gastric adenocarcinomas.METHODS: Methylation-specific PCR (MSP) assay was performed in 89 primary gastric carcinomas (intestinal and diffuse types). Microsatellite instability (MSI) analysis was performed using the BAT26 primer set and PCR products were analyzed with the ABI PRISM 3100 Genetic Analyzer using G...

  2. Study on RIZ1 gene promoter methylation status in human esophageal squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    Shang-Wen Dong; Peng Zhang; Yi-Mei Liu; Yuan-Tao Cui; Shuo Wang; Shao-Jie Liang; Zhun He; Pei Sun; Yuan-Guo Wang

    2012-01-01

    AIM: To investigate the promoter region methylation status of retinoblastoma protein-interacting zinc finger gene 1 (RIZ1) in the human esophageal squamous cell carcinoma (ESCC) cell lines and tissues and verify the relationship between methylation of RIZ1 and oncogen-esis, tumor progression and metastasis etc of ESCC.METHODS: Methylation-specific polymerase chain reaction (MSP) was used to investigate the promoter region methylation status of RIZ1 in 6 ESCC cell lines. One cell line where RIZ1 promoter region methylation was detected was selected for the next study, where the cell line was treated with 5-aza-CdR. Real-time polymerase chain reaction was used to investigate its influence on the transcription of RIZ1. Experiments using frozen pathological specimens from 47 ESCC patients were performed using the same MSP methodology.RESULTS: Promoter methylation of RIZ1 gene was detected in TE13, CaEs17 and EC109 cell lines and the cell line TE13 was chosen for further study. The expression of RIZ1 mRNA in TE-13 was up-regulated after treatment with 5-aza-CdR. The rate of methylation in carcinomas tissues was significantly higher than those in matched neighboring normal and distal ending normal tissue, and the deviation of data was statistically significant (x2 = 24.136, P < 0.01). Analysis of the gender, age familial history, tumour deviation, tumour saturation, lymph gland displacement and clinical staging of 47 samples from ESCC patients showed that the fluctuation of data was not statistically significant.CONCLUSION: Promoter methylation may play an important role in the epigenetic silencing of RIZ1 gene expression in human ESCC. RIZ1 is considered to be a potential tumor suppressor gene and may be a biological parameter for testing early stage human ESCC.

  3. Methylation in hepatocellular carcinoma

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    Regina M. Santella

    2007-02-01

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    The development of HCC is a complex, multistep, multistage process. The molecular pathogenesis of HCC appears to involve multiple genetic aberrations in the molecular control of hepatocyte proliferation, differentiation and death and the maintenance of genomic integrity. This process is influenced by the cumulative activation and inactivation of oncogenes, tumor suppressor genes and other genes. p53, a tumor suppressor gene, is the most frequently mutated gene in human cancers. There is also a striking sequence specific binding and induction of mutations by AFB1 at codon 249 of p53 in HCC.

    Epigenetic alterations are also involved in cancer development and progression. Methylation of promoter CpG islands is associated with inhibition of transcriptional initiation and permanent silencing of downstream genes.

    It is now known that most important tumor suppressor genes are inactivated, not only by mutations and deletions but also by promoter methylation. Several studies indicated that p16, p15, RASSF1A, MGMT, and GSTP1 promoter hypermethylation are prevalent in HCC. In addition, geographic variation in the methylation status of tumor DNA indicates that environmental factors may influence the frequent and concordant degree of hypermethylation in multiple genes in HCC and that epigeneticenvironmental interactions may be involved in hepatocarcinogenesis. We have found significant relationships between promoter methylation and AFB1-DNA adducts confirming the impact of environmental exposures on gene methylation.

    DNA isolated from serum or plasma of cancer patients frequently contains the same genetic and

  4. DNA methylation in hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Iris Tischoff; Andrea Tannapfel

    2008-01-01

    As for many other tumors, development of hepatocellular carcinoma (HCC) must be understood as a multistep process with accumulation of genetic and epigenetic alterations in regulatory genes, leading to activation of oncogenes and inactivation or loss of tumor suppressor genes (TSG). In the last decades, in addition to genetic alterations, epigenetic inactivation of (tumor suppressor) genes by promoter hypermethylation has been recognized as an important and alternative mechanism in tumorigenesis. In HCC, aberrant methylation of promoter sequences occurs not only in advanced tumors, it has been also observed in premalignant conditions just as chronic viral hepatitis B or C and cirrhotic liver. This review discusses the epigenetic alterations in hepatocellular carcinoma focusing DNA methylation.

  5. Study on the polymorphisms and promoter methylation and expression of the glutathione Stransferases P1 gene in hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    张友才

    2006-01-01

    Objective To study the relationship between hepatocellular carcinoma (HCC) and the polymorphisms, promoter methylation, and expression of glutathione S-transferases P1 gene (GST)P1 gene. Methods Using methylation -special PCR (MSP), the methylated status of CpG islands of GSTP1 gene in tumor tissues of 53 HCC and its adjacent nontumor tissues were studied. The en-

  6. Evaluation of MYB Promoter Methylation in Salivary Adenoid Cystic Carcinoma

    Science.gov (United States)

    Shao, Chunbo; Bai, Weiliang; Junn, Jacqueline C.; Uemura, Mamoru; Hennessey, Patrick T.; Zaboli, David; Sidransky, David; Califano, Joseph A.; Ha, Patrick K.

    2011-01-01

    Summary The transcription factor MYB was recently proposed to be a promising oncogene candidate in salivary gland adenoid cystic carcinoma (ACC). However, the up-regulation of MYB in ACC could not be explained solely by deletion of its 3′ end. It is widely accepted that the promoter methylation status can regulate the transcription of genes, especially in human cancers. Therefore, it is important to know whether MYB promoter demethylation could explain the over-expression of MYB in ACC. By using the Methprimer program, we identified nine CpG islands in the promoter of MYB. All of these CpG islands were located within the −864 to +2,082 nt region relative to the transcription start site of MYB. We then used bisulfite genomic sequencing to evaluate the methylation levels of the CpG islands of MYB in 18 primary ACC tumors, 13 normal salivary gland tissues and nine cancer cell lines. Using cell lines, we also determined the relative MYB expression levels and correlated these with the methylation levels. With bisulfite genomic sequencing, we found no detectable methylation in the CpG islands of MYB in either ACC or normal salivary gland tissues. There was a variable degree of MYB expression in the cell lines tested, but none of these cell lines demonstrated promoter methylation. Promoter hypomethylation does not appear to explain the differential expression of MYB in ACC. An alternative mechanism needs to be proposed for the transcriptional control of MYB in ACC. PMID:21324728

  7. MicroRNA-137 promoter methylation in oral lichen planus and oral squamous cell carcinoma

    DEFF Research Database (Denmark)

    Dang, Jun; Bian, Yong-qian; Sun, Jian-yong

    2013-01-01

    Oral lichen planus (OLP) is a common oral mucosal disease, which is generally considered a potentially malignant lesion. To identify efficiently prognostic biomarker, we investigated the microRNA-137 (miR-137) promoter methylation in OLP and compared with the samples from healthy volunteers...... and patients with oral squamous cell carcinoma (OSCC). A total of 20 OLP and 12 patients with OSCC as well as 10 healthy subjects were subjected to miR-137 promoter methylation analysis using methylation-specific PCR (MSP). To address the malignancy prediction potential from miR-137 promoter methylation status...... between miR-137 and p16 methylation levels were statistically significant between healthy controls and patients. Methylation levels of the two promoters were also influenced by age, gender, and lesion duration. Interestingly, aberrant promoter methylation of the p16 and miR-137 genes was only found...

  8. Frequent and distinct aberrations of DNA methylation patterns in fibrolamellar carcinoma of the liver.

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    Wolfgang Tränkenschuh

    Full Text Available BACKGROUND: Gene silencing due to aberrant DNA methylation is a frequent event in hepatocellular carcinoma (HCC and also in hepatocellular adenoma (HCA. However, very little is known about epigenetic defects in fibrolamellar carcinoma (FLC, a rare variant of hepatocellular carcinoma that displays distinct clinical and morphological features. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed the methylation status of the APC, CDH1, cyclinD2, GSTπ1, hsa-mir-9-1, hsa-mir-9-2, and RASSF1A gene in a series of 15 FLC and paired normal liver tissue specimens by quantitative high-resolution pyrosequencing. Results were compared with common HCC arising in non-cirrhotic liver (n = 10. Frequent aberrant hypermethylation was found for the cyclinD2 (19% and the RASSF1A (38% gene as well as for the microRNA genes mir-9-1 (13% and mir-9-2 (33%. In contrast to common HCC the APC and CDH1 (E-cadherin genes were found devoid of any DNA methylation in FLC, whereas the GSTπ1 gene showed comparable DNA methylation in tumor and surrounding tissue at a moderate level. Changes in global DNA methylation level were measured by analyzing methylation status of the highly repetitive LINE-1 sequences. No evidence of global hypomethylation could be found in FLCs, whereas HCCs without cirrhosis showed a significant reduction in global methylation level as described previously. CONCLUSIONS: FLCs display frequent and distinct gene-specific hypermethylation in the absence of significant global hypomethylation indicating that these two epigenetic aberrations are induced by different pathways and that full-blown malignancy can develop in the absence of global loss of DNA methylation. Only quantitative DNA methylation detection methodology was able to identify these differences.

  9. Aberrant DNA methylation associated with MTHFR C677T genetic polymorphism in cutaneous squamous cell carcinoma in renal transplant patients.

    LENUS (Irish Health Repository)

    Laing, M E

    2010-08-01

    Changes in genomic DNA methylation associated with cancer include global DNA hypomethylation and gene-specific hyper- or hypomethylation. We have previously identified a genetic variant in the MTHFR gene involved in the methylation pathway which confers risk for the development of squamous cell carcinoma (SCC) in renal transplant patients. This genetic variant has also been discovered to confer SCC risk in nontransplant patients with low folate status.

  10. CMTM5 exhibits tumor suppressor activity through promoter methylation in oral squamous cell carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Heyu [Central Laboratory, Peking University School of Stomatology, Beijing (China); Nan, Xu [Center for Human Disease Genomics, Department of Immunology, Key Laboratory of Medical Immunology, Ministry of Health, School of Basic Medical Sciences, Peking University, Beijing (China); Li, Xuefen [Central Laboratory, Peking University School of Stomatology, Beijing (China); Chen, Yan; Zhang, Jianyun [Department of Oral Pathology, Peking University School of Stomatology, Beijing (China); Sun, Lisha [Central Laboratory, Peking University School of Stomatology, Beijing (China); Han, Wenlin [Center for Human Disease Genomics, Department of Immunology, Key Laboratory of Medical Immunology, Ministry of Health, School of Basic Medical Sciences, Peking University, Beijing (China); Li, Tiejun, E-mail: litiejun22@vip.sina.com [Department of Oral Pathology, Peking University School of Stomatology, Beijing (China)

    2014-05-02

    Highlights: • Down-regulation of CMTM5 expression in OSCC tissues was found. • The promoter methylation status of CMTM5 was measured. • CMTM5-v1 inhibited cell proliferation and migration and induced apoptosis. • CMTM5 might act as a putative tumor suppressor gene in OSCC. - Abstract: Oral squamous cell carcinoma (OSCC) is one of the most common types of malignancies in the head and neck region. CKLF-like MARVEL transmembrane domain-containing member 5 (CMTM5) has been recently implicated as a tumor suppressor gene in several cancer types. Herein, we examined the expression and function of CMTM5 in oral squamous cell carcinoma. CMTM5 was down-regulated in oral squamous cell lines and tumor samples from patients with promoter methylation. Treatment with the demethylating agent 5-aza-2′-deoxycytidine restored CMTM5 expression. In the OSCC cell lines CAL27 and GNM, the ectopic expression of CMTM5-v1 strongly inhibited cell proliferation and migration and induced apoptosis. In addition, CMTM5-v1 inhibited tumor formation in vivo. Therefore, CMTM5 might act as a putative tumor suppressor gene through promoter methylation in oral squamous cell carcinoma.

  11. Study on MXR7 methylation in hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    GUO Ting-ting; ZENG Jin-zhang; WANG Hong-yang

    2005-01-01

    Objective:To obtain information at the molecular level on the possible mechanism of MXR7 gene overexpressed in hepatocellular carcinoma (HCC) and also to provide a clue for further study. Methods:Genomic DNA was isolated from 20 samples of hepatoma and paired non-HCC liver tissues, 2 cases of blood tumor and two types of cells (HepG2, MCF-7) and digested with two kinds of endonucleases (EcoR Ⅰ and EagⅠwhich is methylation sensitive endonuclease). And the condition of MXR7 gene methylation was examined and analyzed by Southern blot. Results: MXR7 was unmethylated neither in tested tumorous liver samples nor in paired non-HCC liver tissues. In addition, the same result was found in 2 blood tumor samples and HepG2. Only two paired samples had different methylation outcome, one was unmethylated and the other was partly methylated. Conclusion: MXR7 is unmethylated in HHC, suggesting methylation of MXR7 may have no relation with its expression and regulation.

  12. Plasma DNA methylation of Wnt antagonists predicts recurrence of esophageal squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    Ji-Bin Liu; Fu-Lin Qiang; Jing Dong; Jin Cai; Shu-Hui Zhou; Min-Xin Shi; Ke-Ping Chen; Zhi-Bin Hu

    2011-01-01

    AIM: To detect the effects of plasma DNA methylation of Wnt antagonists/inhibitors on recurrence of esophageal squamous cell carcinoma (ESCC).METHODS: We used methylation-specific polymerase chain reaction to detect hypermethylation of the promoter of four Wnt antagonists/inhibitors (SFRP-1, WIF-1, DKK-3 and RUNX3) using DNA from the plasma of ESCC patients (n = 81) and analyzed the association between promoter hypermethylation of Wnt pathway modulator genes and the two-year recurrence of ESCC.RESULTS: Hypermethylation of SFRP-1, DKK-3 and RUNX-3 was significantly associated with an increased risk of ESCC recurrence (P = 0.001, 0.003 and 0.001 for SFRP-1, DKK-3 and RUNX3, respectively). Patients carrying two to three methylated genes had a significantly elevated risk of recurrence compared with those not carrying methylated genes (odds ratio = 15.69, 95% confidential interval: 2.97-83). The area under the receiver operating characteristic curve (AUC) was 77.1 for ESCC recurrence prediction (sensitivity = 66.67 and specificity = 83.3). When combining methylated genes and the clinical stage, the AUC was 83.69, with a sensitivity of 76.19 and a specificity of 83.3.CONCLUSION: The status of promoter hypermethylation of Wnt antagonists/inhibitors in plasma may serve as a non-invasive prognostic biomarker for ESCC.

  13. Prognostic significance of CpG island methylator phenotype in surgically resected small cell lung carcinoma.

    Science.gov (United States)

    Saito, Yuichi; Nagae, Genta; Motoi, Noriko; Miyauchi, Eisaku; Ninomiya, Hironori; Uehara, Hirofumi; Mun, Mingyon; Okumura, Sakae; Ohyanagi, Fumiyoshi; Nishio, Makoto; Satoh, Yukitoshi; Aburatani, Hiroyuki; Ishikawa, Yuichi

    2016-03-01

    Methylation is closely involved in the development of various carcinomas. However, few datasets are available for small cell lung cancer (SCLC) due to the scarcity of fresh tumor samples. The aim of the present study is to clarify relationships between clinicopathological features and results of the comprehensive genome-wide methylation profile of SCLC. We investigated the genome-wide DNA methylation status of 28 tumor and 13 normal lung tissues, and gene expression profiling of 25 SCLC tissues. Following unsupervised hierarchical clustering and non-negative matrix factorization, gene ontology analysis was performed. Clustering of SCLC led to the important identification of a CpG island methylator phenotype (CIMP) of the tumor, with a significantly poorer prognosis (P = 0.002). Multivariate analyses revealed that postoperative chemotherapy and non-CIMP were significantly good prognostic factors. Ontology analyses suggested that the extrinsic apoptosis pathway was suppressed, including TNFRSF1A, TNFRSF10A and TRADD in CIMP tumors. Here we revealed that CIMP was an important prognostic factor for resected SCLC. Delineation of this phenotype may also be useful for the development of novel apoptosis-related chemotherapeutic agents for treatment of the aggressive tumor.

  14. Methylation of IGF2 regulatory regions to diagnose adrenocortical carcinomas.

    Science.gov (United States)

    Creemers, S G; van Koetsveld, P M; van Kemenade, F J; Papathomas, T G; Franssen, G J H; Dogan, F; Eekhoff, E M W; van der Valk, P; de Herder, W W; Janssen, J A M J L; Feelders, R A; Hofland, L J

    2016-09-01

    Adrenocortical carcinoma (ACC) is a rare malignancy with a poor prognosis. Discrimination of ACCs from adrenocortical adenomas (ACAs) is challenging on both imaging and histopathological grounds. High IGF2 expression is associated with malignancy, but shows large variability. In this study, we investigate whether specific methylation patterns of IGF2 regulatory regions could serve as a valuable biomarker in distinguishing ACCs from ACAs. Pyrosequencing was used to analyse methylation percentages in DMR0, DMR2, imprinting control region (ICR) (consisting of CTCF3 and CTCF6) and the H19 promoter. Expression of IGF2 and H19 mRNA was assessed by real-time quantitative PCR. Analyses were performed in 24 ACCs, 14 ACAs and 11 normal adrenals. Using receiver operating characteristic (ROC) analysis, we evaluated which regions showed the best predictive value for diagnosis of ACC and determined the diagnostic accuracy of these regions. In ACCs, the DMR0, CTCF3, CTCF6 and the H19 promoter were positively correlated with IGF2 mRNA expression (P<0.05). Methylation in the most discriminating regions distinguished ACCs from ACAs with a sensitivity of 96%, specificity of 100% and an area under the curve (AUC) of 0.997±0.005. Our findings were validated in an independent cohort of 9 ACCs and 13 ACAs, resulting in a sensitivity of 89% and a specificity of 92%. Thus, methylation patterns of IGF2 regulatory regions can discriminate ACCs from ACAs with high diagnostic accuracy. This proposed test may become the first objective diagnostic tool to assess malignancy in adrenal tumours and facilitate the choice of therapeutic strategies in this group of patients.

  15. Aberrant methylation of Glutathione S-transferase P1 and E-cadherin in invasive ductal breast carcinoma and fibroadenoma

    Institute of Scientific and Technical Information of China (English)

    Wings Tjing Yung Loo; Mary Ngan Bing Cheung; Louis Wing Cheong Chow

    2010-01-01

    Objective To investigate the hypermethylation status of glutathione transferase P1(GSTP1) and E cadherin (ECAD), TSGs (tumor suppressor genes) in our breast cancer samples and explore their correlation with clinicopathological features of corresponding cancer patients. Methods One hundred and thirty six IDC (invasive ductal carcinoma) patients were recruited for analysis and 16 fibroadenoma patients acted as control. DNA extraction and methylation specific PCR (MSP) were subsequently performed preceded by pathological examination. Results The percentage of hypermethylated GSTP1 in carcinoma and fibroadenoma groups was 34.92% and 15.79% respectively and the percentage of hypermethylated ECAD in carcinomas and fibroadenomas was 18.00% and 0.00% respectively. Carcinoma had the highest percentage of c erbB2 overexpression being 54.55% among the clinicopathological parameters. Conclusion Hypermethylation patterns are frequent in IDC and seem to relate to c erbB2 overexpression, and such epigenetic change should not be neglected in fibroadenoma. Tumor methylation status in cancer patients can be determined at early stage and it may be a reference for better treatment planning.

  16. Status of hepatocellular carcinoma in Gulf region.

    Science.gov (United States)

    Rasul, Kakil Ibrahim; Al-Azawi, Safaa H; Chandra, Prem; Abou-Alfa, Ghassan K; Knuth, Alexander

    2013-12-01

    Hepatocellular carcinoma (HCC) has a unique geographic distribution that is likely to be determined by specific etiologic factors. There is a distinctive difference in sex and age related occurrence of disease. In the Gulf region, there are contradicting data on the prevalence and death rates due to HCC. In this review we highlight some aspects of HCC specific to the Gulf region. A retrospective analysis of 150 patient's data is presented, including demographic, epidemiological, aetiological disease status assessment with child Pugh criteria, modes of treatment and treatment related outcome. Hepatitis C virus (HCV) infection was the most common (45%) documented etiology, similar to Western European countries, followed by hepatitis B virus (HBV) infection in 27% of cases, alcoholic liver disease only in six patients (4%). Child-Pugh assessment was A in 33%, B in 37% and C in 30% of observed patients. Surgery (liver resection or transplantation) was performed in 12% and local ablation in 5% of cases. The others were treated by chemo-embolization in 17% and by systemic therapy with sorafenib in 13% of patients. Nearly half of the patients (53%) were in advanced stages and received palliative treatment. To improve the outcome of treatment in HCC patients in the Gulf region, an effective and strategic screening program must be implemented for early diagnosis and treatment to improve the outcome of this mostly fatal disease.

  17. Effects of DNMT3b on expression and methylation status of promoter region of DLC-I in human hepatocellular carcinoma cell line%DNMT3b对肝癌细胞株中DLC-1的表达及启动子区甲基化状况的影响

    Institute of Scientific and Technical Information of China (English)

    王家祥; 刘怀然; 范正军

    2008-01-01

    目的 探讨DNA甲基化转移酶3b(DNMT3b)对人肝癌细胞株SMMC-7721中DLC-1基因的表达及其启动子区甲基化状况的影响.方法 将SMMC-7721细胞株分为两组,试验组应用siRNA技术沉默DNMT3b的表达,对照组仅转染对照siRNA;应用Western Blot技术分别榆测两组细胞中DNMT3b和DLC-1的表达,并应用甲基化特异性PCR(MSP)技术分别检测两组细胞中DLC-1基因启动子区的甲基化状况.结果 试验组中DNMT3b的表达明显低于对照组,而DLC-1的表达明显高于对照组;两组中DLC-1启动子区甲基化状态无差异,均发生甲基化.结论 siRNA技术抑制DNMT3b的表达可使DLC-1基因表达水平增高,但DLC-1启动子区甲基化状态无变化.此过程中DNMT3b并非作为甲基转移酶,而可能是作为转录调控因子影响DLC-1的表达.%Objective To explore the effects of DNMT3b on the expression and methylation sta-tus of the promoter region of DLC-1 in human hepatocellular carcinoma cell line. Methods The SMMC-7721 cell line was divided into 2 groups. The cell line in the experimental group was transfect-ed with DNMT3b siRNA, while that in the control group was transfected with control siRNA. West-ern blot was used to detect the expression of DNMT3b and DLC-1 and MSP was employed to examine the methylation status of the promoter region of DLC-1. Results The expression of DNMT3b was significantly higher in the experimental group than in the control group, while the expression of DLC-1 was just opposite. There was no significant difference in the methylation status of the promoter re-gion of DLC-1 between the 2 groups and both were methylated. Conelnsion The inhibition of expression of DNMT3b by siRNA method can enhance the expression level of DLC-1, and the methylation status of the promoter region of DLC-1 does not change at the same time. When affecting the expression of DLC-1, DN-MT3b might not play the role of methyhransferase, but can act as a transcriptional regulatory

  18. Genome-wide methylation analysis of tubulocystic and papillary renal cell carcinomas.

    Science.gov (United States)

    Korabecna, M; Geryk, J; Hora, M; Steiner, P; Seda, O; Tesar, V

    2016-01-01

    Tubulocystic renal cell carcinoma (TRCC) represents a rare tumor with incidence lower than 1 % of all renal carcinomas. This study was undertaken to contribute to characterization of molecular signatures associated with TRCC and to compare them with the features of papillary renal cell carcinoma (PRCC) at the level of genome wide methylation analysis.We performed methylated DNA immunoprecipitation (MeDIP) coupled with microarray analysis (Roche NimbleGen). Using the CHARM package, we compared the levels of gene methylation between paired samples of tumors and control renal tissues of each examined individual. We found significant global demethylation in all tumor samples in comparison with adjacent kidney tissues of normal histological appearance but no significant differences in gene methylation between the both compared tumor entities. Therefore we focused on characterization of differentially methylated regions between both tumors and control tissues. We found 42 differentially methylated genes.Hypermethylated genes for protocadherins (PCDHG) and genes coding for products associated with functions of plasma membrane were evaluated as significantly overrepresented among hypermethylated genes detected in both types of renal cell carcinomas.In our pilot study, we provide the first evidence that identical features in the process of carcinogenesis leading to TRCC and/or to PRCC may be found at the gene methylation level.

  19. CpG island methylator phenotype in plasma is associated with hepatocellular carcinoma prognosis

    Institute of Scientific and Technical Information of China (English)

    Ji-Bin Liu; Yi-Xin Zhang; Shu-Hui Zhou; Min-Xin Shi; Jin Cai; Yan Liu; Ke-Ping Chen; Fu-Lin Qiang

    2011-01-01

    AIM: To evaluate the clinical significance of CpG island methylator phenotype (CIMP) in plasma and its asso ciation with hepatocellular carcinoma (HCC) progress.METHODS: CIMP status of 108 HCC patients was analyzed using a methylation marker panel in tumor tissues and plasma with methylation-specific poly merase chain reaction. Fifteen samples of non-neo plastic liver tissues and 60 of plasma from healthy persons were examined simultaneously. Examined genes included APC, WIF-1, RUNX-3, DLC-1, SFRP-1, DKK and E-cad.RESULTS: The frequencies of high-level methyla tion in HCC tissue and plasma were at least 15% for the seven genes: APC, 48/108, 44.44% in tissue and 26/108, 24.07% in plasma; WIF-1, 53/108, 49.07% in tissue and 35/108, 32.41% in plasma; RUNX-3, 52/108, 48.14% in tissue and 42/108, 38.89% in plasma; DLC-1, 38/108, 35.18% in tissue and 23/108, 21.30% in plasma; SFRP-1, 40/108, 37.04% in tissue and 31/108, 28.7% in plasma; DKK, 39/108, 36.1% in tis sue and 25/108, 23.14% in plasma; and E-cad, 37/108, 34.3% in tissue and 18/108, 16.67% in plasma. CIMP+ (S 3 methylated genes) was detected in 68 (60.2%) tumor tissue samples and 62 (57.4%) plasma samples. CIMP was not detected in non-neoplastic liver tissues or plasma of healthy persons. CIMP status in tumor tissues differed significantly in gender, hepatitis B surface antigen, alpha-fetoprotein, and tumor-node-metastasis stage (P < 0.05). Similar results were ob tained with plasma samples (P < 0.05). There was no difference in CIMP status in age, presence of hepatitis C virus antibody, cirrhosis, number of nodes, number of tumors, tumor size, or Edmondson-Steiner stage. A one-year follow-up found that the metastatic rate and recurrence rate in the CIMP+ group were significantly higher than in the CIMP- group as assessed with plas ma samples (P < 0.05).CONCLUSION: Plasma DNA can be a reliable sample source for CIMP analysis. CIMP in plasma may serve as a molecular marker of late-stage and poor-prognosis HCC.

  20. Microarray-based method for detecting methylation changes of p16Ink4a gene 5'-CpG islands in gastric carcinomas

    Institute of Scientific and Technical Information of China (English)

    Peng Hou; Jia-Yao Shen; Mei-Ju Ji; Nong-Yue He; Zu-Hong Lu

    2004-01-01

    AIM: Aberrant DNA methylation of CpG site is among the earliest and most frequent alterations in cancer. Several studies suggest that aberrant methylation of the CpG sites of the tumor suppressor gene is closely associated with carcinogenesis. However, large-scale analysis of candidate genes has so far been hampered by the lack of highthroughput approach for analyzing DNA methylation. The aim of this study was to describe a microarray-based method for detecting changes of DNA methylation in cancer.METHODS: This method used bisulfite-modified DNA as a template for PCR amplification, resulting in conversion of unmethylated cytosine, but not methylated cytosine, into thymine within CpG islands of interest. Therefore, the amplified product might contain a pool of DNA fragments with altered nucleotide sequences due to differential methylation status.Nine sets of oligonucleotide probes were designed to fabricate a DNA microarray to detect the methylation changes of p16 gene CpG islands in gastric carcinomas. The results were further validated by methylation-specific PCR (MSP).RESULTS: The experimental results showed that the microarray assay could successfully detect methylation changes of p16 gene in 18 gastric tumor samples. Moreover,it could also potentially increase the frequency of detecting p16 methylation from tumor samples than MSP.CONCLUSION: Microarray assay could be applied as a useful tool for mapping methylation changes in multiple CpG loci and for generating epigenetic profiles in cancer.

  1. Disordered beta-catenin expression and E-cadherin/CDH1 promoter methylation in gastric carcinoma

    Institute of Scientific and Technical Information of China (English)

    Li Wang; Fan Zhang; Ping-Ping Wu; Xu-Cheng Jiang; Lin Zheng; Ying-Yan Yu

    2006-01-01

    AIM: To investigate the distribution of beta-catenin in nuclei or membrane/cytoplasm of gastric carcinoma cells,the relationship between E-cadherin gene methylation and its expression, and the role of beta-catenin and E-cadherin as potential molecular markers in predicting tumor infiltration.METHODS: Twenty-nine cases of gastric carcinoma,classified as diffuse and intestinal variants, were selected for study. Nuclear and cytoplasmic proteins were purified and beta-catenin content was detected by ELISA. DNA methylation of E-cadherin/CDH1 gene promoter was studied by methylation-specific PCR and compaired with E-cadherin expression detected by immunohistochemistry.RESULTS: In 27 cases of gastric carcinoma, the ratio of beta-catenin content between nuclei and membrane/cytoplasm was correlated with the T-classification (r =0.392, P = 0.043). The significance was present between T2 and T3 groups. No correlation was detected between diffuse and intestinal variants in terms of their betacatenin distribution. In 21 cases of diffuse variants of gastric carcinoma, there was a difference in E-cadherin expression between CDH1 gene-methylated group and non-methylated group (29 % vs 71%, P = 0.027).No correlation between CDH1 gene methylation and T-classification was found, neither was the significance between E-cadherin expression and tumor infiltration grade.CONCLJSION: Comparative analysis of nuclear and membrane/cytoplasmic beta-catenin can predict local tumor infiltration. E-cadherin/CDH1 gene methylation is an important cause for its gene silence in diffuse variant gastric carcinoma. Methylation of CDH1 gene in the absence of E-cadherin is an early event in gastric carcinogenesis.

  2. Urticaria after methyl aminolevulinate photodynamic therapy in a patient with nevoid basal cell carcinoma syndrome.

    Science.gov (United States)

    Wolfe, Christopher M; Green, W Harris; Hatfield, H Keith; Cognetta, Armand B

    2012-11-01

    Methyl aminolevulinate photodynamic therapy (MAL-PDT) is utilized in several countries for the treatment of basal cell carcinoma, but allergic sensitization has been reported by the manufacturer. To the best of our knowledge, we report the first case of urticaria following MAL-PDT in a patient with nevoid basal cell carcinoma syndrome. Prophylactic use of antihistamines may allow continued use of MAL-PDT in this setting.

  3. p16/MTS1 inactivation in ovarian carcinomas: high frequency of reduced protein expression associated with hyper-methylation or mutation in endometrioid and mucinous tumors.

    Science.gov (United States)

    Milde-Langosch, K; Ocon, E; Becker, G; Löning, T

    1998-02-20

    Inactivation of the tumor-suppressor gene p16 (MTS1/ CDKN2/INK4a) has been described in various human malignancies. Although p16 deletion has been found in various ovarian tumor cell lines, p16 inactivation by homozygous deletion or mutation has been reported only sporadically in primary ovarian carcinomas. In a comprehensive study, we analyzed p16 protein expression by immuno-histochemistry (IHC) on paraffin sections of 94 primary ovarian carcinomas of different histological subtype. Loss of expression was detected in 19 primary tumors (20%), mainly mucinous and endometrioid carcinomas. To reveal the cause of suppressed expression, we performed (i) analysis of homozygous deletions by comparative PCR after micro-dissection, (ii) mutation analysis by single-strand conformation polymorphism analysis and subsequent direct sequencing and (iii) methylation-specific PCR to determine the methylation status of 5'-CpG islands. Loss of or weak p16 expression was caused by hyper-methylation (12/19 IHC-negative cases), somatic mutation (10 tumors) or homozygous deletion (1 case). Aberrant p 16 results by one of these methods were detected in 71-79% of endometrioid and mucinous, but in only 10% of serous-papillary, carcinomas. Our data suggest that p16 inactivation is a typical feature of certain subtypes of ovarian carcinoma.

  4. Genome-wide DNA methylation profiling of non-small cell lung carcinomas

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    Carvalho Rejane

    2012-06-01

    Full Text Available Abstract Background Non-small cell lung carcinoma (NSCLC is a complex malignancy that owing to its heterogeneity and poor prognosis poses many challenges to diagnosis, prognosis and patient treatment. DNA methylation is an important mechanism of epigenetic regulation involved in normal development and cancer. It is a very stable and specific modification and therefore in principle a very suitable marker for epigenetic phenotyping of tumors. Here we present a genome-wide DNA methylation analysis of NSCLC samples and paired lung tissues, where we combine MethylCap and next generation sequencing (MethylCap-seq to provide comprehensive DNA methylation maps of the tumor and paired lung samples. The MethylCap-seq data were validated by bisulfite sequencing and methyl-specific polymerase chain reaction of selected regions. Results Analysis of the MethylCap-seq data revealed a strong positive correlation between replicate experiments and between paired tumor/lung samples. We identified 57 differentially methylated regions (DMRs present in all NSCLC tumors analyzed by MethylCap-seq. While hypomethylated DMRs did not correlate to any particular functional category of genes, the hypermethylated DMRs were strongly associated with genes encoding transcriptional regulators. Furthermore, subtelomeric regions and satellite repeats were hypomethylated in the NSCLC samples. We also identified DMRs that were specific to two of the major subtypes of NSCLC, adenocarcinomas and squamous cell carcinomas. Conclusions Collectively, we provide a resource containing genome-wide DNA methylation maps of NSCLC and their paired lung tissues, and comprehensive lists of known and novel DMRs and associated genes in NSCLC.

  5. Genome-wide DNA methylation profiling of non-small cell lung carcinomas

    NARCIS (Netherlands)

    R.H. Carvalho (Rejane Hughes); V. Haberle (Vanja); J. Hou (Jun); T. van Gent (Teus); S. Thongjuea (Supat); W.F.J. van IJcken (Wilfred); C. Kockx (Christel); R.W.W. Brouwer (Rutger); E.J. Rijkers; A.M. Sieuwerts (Anieta); J.A. Foekens (John); M. van Vroonhoven (Mirjam); J.G.J.V. Aerts (Joachim); F.G. Grosveld (Frank); B. Lenhard (Boris); J.N.J. Philipsen (Sjaak)

    2012-01-01

    textabstractBackground: Non-small cell lung carcinoma (NSCLC) is a complex malignancy that owing to its heterogeneity and poor prognosis poses many challenges to diagnosis, prognosis and patient treatment. DNA methylation is an important mechanism of epigenetic regulation involved in normal developm

  6. Detection of methylation status of promoter region of CASR in cell and tissue of human esophageal squamous cell carcinoma%食管鳞状细胞癌细胞及组织中钙敏感受体基因启动子区甲基化状态检测

    Institute of Scientific and Technical Information of China (English)

    景坤玉; 董腾慧; 郭明洲; 薛乐勋

    2013-01-01

    Aim:To investigate the promoter region methylation status of calcium-sensing receptor( CASR ) in esophageal carcinogenesis, and to explore the possibility of CASR promoter methylation as a potential biomarker for human esophageal squamous cell carcinoma ( ESCC ). Methods:The promoter region methylation of CASR in 6 esophageal cancer cell lines, 8 normal esophageal mucosa and 68 primary ESCC tissue samples were investigated using methylation specific PCR. Semi-quantitative RT-PCR was used to evaluate CASR expression levels before and after treatment with 5-aza-2'-deoxycyti-dine ( 5-Aza ) in cell lines. Results:Methylation of the CASR promoter region was detected in 5 cell lines( KYSE30, KYSE70, KYSE140.TE8 and BIC1 ), but not in KYSE510. Loss of CASR expression was found in the methylated cell lines. Re-expression of CASR was induced after 5-Aza treatment for 96 h. In 68 cases of human primary ESCC tissue samples, the methylation rate of CASR promoter region was 71% ( 48/68 ), but no one was methylated in 8 cases of normal e-sophageal mucosa( 0/8 ). No association was found between promoter region hypermethylation and age(X2= 1. 114, P = 0.291 ), gender(X2=0.342, P =0.558 ), tumor location(X2= 1. 209 , P =0. 546 ), tumor stage (X2 =0.181, P= 0.670) or lymph node metastasis(X2= 1. 169, P =0. 280 ). Conclusion:The methylated CASR frequently exists in primary human ESCC, suggesting it may be used as a potential early detection biomarker or target for this disease.%目的:研究钙敏感受体(CASR)基因启动子区甲基化状态与食管鳞状细胞癌(ESCC)发生之间的关系,探讨CASR甲基化作为潜在的ESCC早期诊断标志物的可能性.方法:采用甲基化特异性PCR(MSP)方法分析6株食管癌细胞系、8例正常食管黏膜组织以及68例原发ESCC组织中CASR启动子区的甲基化状态.采用半定量RT-PCR方法分析甲基化酶抑制剂5-氮杂-2-脱氧胞嘧啶核苷(5-Aza)处理前后食管癌细胞系中CASR m

  7. GSTP1 expression and promoter methylation in epithelial ovarian carcinoma

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    V Shilpa

    2014-01-01

    Full Text Available Context: GSTP1 is a subgroup of glutathione-S-transferase family, which provides cellular protection against free radical and carcinogenic compounds due to its detoxifying function. Altered GSTP1 activity due to down regulation of enzyme activity and DNA methylation has been reported in many tumors, although data for ovarian cancer are few. In this study, we aimed at determining the expression of GSTP1 in relation to the methylation of the GSTP1 promoter in epithelial ovarian cancer (EOC. Materials and Methods: GSTP1 mRNA expression and GSTP1 enzyme concentration were assessed by quantitative reverse transcriptase polymerase chain reaction (PCR and enzyme-linked immunosorbent assay, respectively, in 88 EOCs, 14 low malignant potential (LMP tumors, and 20 benign tumors. The promoter methylation of GSTP1 gene was evaluated by methylation-specific PCR. Results: Reduced GSTP1 mRNA expression was observed in 49% EOCs, 21.4% LMP, and 45% benign tumors. Significantly lower levels of plasma GSTP1 were observed in all tumor samples compared to normal. GSTP1 promoter methylation was detected in 10 (11.4% EOCs and 1 (7.3% LMP tumors. No methylation was observed in benign tumors and normal ovaries. Conclusions: Our results show that there is a significant down regulation of GSTP1 expression while hypermethylation of the GSTP1 gene promoter is not very frequent in EOC. Further studies are needed to study underlying mechanisms leading to decreased expression.

  8. Aberrant promoter methylation and gene expression of H-cadherin gene is associated with tumor progression and recurrence in epithelial ovarian carcinoma

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    Rahul Bhagat

    2014-01-01

    Full Text Available Background: Loss of expression of cadherins by promoter hypermethylation has been described in many epithelial cancers, and it may play a role in tumor cell invasion and metastasis. Previously, we reported that E-cadherin gene is frequently methylated in epithelial ovarian cancer. Aim: The aim of this study was to compare the promoter hypermethylation of H-cadherin gene in ovarian epithelial neoplasms to better understand the role of epigenetic silencing in carcinogenesis. Materials and Methods: We examined the promoter methylation of the H-cadherin gene in 134 epithelial ovarian carcinomas (EOC, 23 low malignant potential (LMP tumors, 26 benign cystadenomas and 15 normal ovarian tissues. Methylation was investigated by methylation specific polymerase chain reaction (MSP and the results confirmed by bisulfite DNA sequencing. Relative gene expression of H-cadherin was done using quantitative reverse transcriptase PCR on 51 EOC cases, 9 LMP tumors, 7 benign cystadenomas with 5 normal ovarian tissues. Results: Aberrant methylation of H-cadherin was present in 20 of 134 (15% carcinoma cases, 2 of 23 (09% LMP tumors and 1 of 26 (4% benign cystadenomas. No methylation was observed in any of the normal ovarian tissues. The mRNA expression level of H-cadherin was significantly down-regulated in EOC and LMP tumors than the corresponding normal tissues, whereas the expression level was normal in benign cystadenomas. A significant correlation of H-cadherin promoter methylation was observed with reduced gene expression in EOC. The prevalence of H-cadherin methylation was associated significantly with stage, histopathological grade, and menopausal status of the patient. H-cadherin methylation also had significant association with recurrence and differentiation of tumor. Conclusion: Our findings suggest an association between H-cadherin methylation, tumor progression and recurrence in EOC.

  9. Methylation associated inactivation of RASSF1A and its synergistic effect with activated K-Ras in nasopharyngeal carcinoma

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    Yu Jing

    2009-12-01

    Full Text Available Abstract Background Epigenetic silencing of tumor suppressor genes associated with promoter methylation is considered to be a hallmark of oncogenesis. RASSF1A is a candidate tumor suppressor gene which was found to be inactivated in many human cancers. Although we have had a prelimilary cognition about the function of RASSF1A, the exact mechanisms about how RASSF1A functions in human cancers were largely unknown. Moreover, the effect of mutated K-Ras gene on the function of RASSF1A is lacking. The aim of this study was to investigate the expression profile and methylation status of RASSF1A gene, and to explore its concrete mechanisms as a tumor suppressor gene in Nasopharyngeal Carcinoma. Methods We examined the expression profile and methylation status of RASSF1A in two NPC cell lines, 38 primary nasopharyngeal carcinoma and 14 normal nasopharyngeal epithelia using RT-PCR and methylated specific PCR(MSP respectively. 5-aza-dC was then added to confirm the correlation between hypermethylation status and inactivation of RASSF1A. The NPC cell line CNE-2 was transfected with exogenous pcDNA3.1(+/RASSF1A plasmid in the presence or absence of mutated K-Ras by liposome-mediated gene transfer method. Flow cytometry was used to examine the effect of RASSF1A on cell cycle modulation and apoptosis. Meanwhile, trypan blue dye exclusion assays was used to detect the effect of RASSF1A transfection alone and the co-transfection of RASSF1A and K-Ras on cell proliferation. Results Promoter methylation of RASSF1A could be detected in 71.05% (27/38 of NPC samples, but not in normal nasopharyngeal epithelia. RASSF1A expression in NPC primary tumors was lower than that in normal nasopharyngeal epithelial (p p p p Conclusion Expression of RASSF1A is down-regulated in NPC due to the hypermethylation of promoter. Exogenous expression of RASSF1A is able to induce growth inhibition effect and apoptosis in tumor cell lines, and this effect could be enhanced by activated

  10. Analysis of the expression of Siit/Robo genes and the methylation status of their promoters in the hetpatocellular carcinoma cell lines%人肝癌细胞系Slit/Robo启动子甲基化状态及基因表达的研究

    Institute of Scientific and Technical Information of China (English)

    郑丹; 刘彬彬; 刘银坤; 康晓楠; 孙璐; 郭坤; 孙瑞霞; 陈洁; 赵燕

    2009-01-01

    目的 研究Slit/Robo信号通路相关基因Slit1、Slit2、Slit3和Robo1、Robo3在多种人肝癌细胞系中的表达及甲基化状态,探讨与肝癌发生和发展的关系. 方法提取9种人肝细胞癌细胞株(Hep3B、HepG2、PLC/PRF/5/PRF/5、SMMC-7721、BEL-7402、MHCC97-H,MHCC97-L、LM3、LM6)及对照细胞株L02的基因组DNA和总RNA,采用半定量逆转录聚合酶链反应技术和甲基化特异性聚合酶链反应技术检测Slit1、Slit2,Slit3和Robo1,Robo3基因的基因表达水平与启动子甲基化状态.实验数据应用Paired t检验. 结果 Slit1、Slit2、Slit3基因除个别细胞株外,在不同转移潜能细胞株中均发生了DNA甲基化,同时Slit1和Slit3在mRNA水平几乎均不表达,Slit2基因表达程度在不同转移潜能的细胞株之间存在差异,随着转移潜能的增加表达大致呈下降趋势.作为Slit2受体的Robo1基因在10株肝癌细胞株中均发生甲基化修饰,但除在SMMC-7721、BEL-7402、L02不表达外,其余7种细胞株均有表达.Robo3基因相关CpG岛在9种肝癌细胞株中均未发生甲基化,同时其在mRNA水平均无表达. 结论 Slit/Robo可能在肝癌发生和发展中发挥作用.而Robo3则在肝癌中不发生表达而且其表达沉默可能不受甲基化方式调控.%Objective To analyze the expression of genes in the Slit/Robo signaling pathway, and the methylation status of their promoters in hetpatocellular carcinoma (HCC) cell lines. Methods Genomic DNA and total RNA were isolated from 9 HCC cell lines of different metastatic ability (Hep3B, HepG2, PLC/PRF/5, SMMC-7721, BEL-7402, MHCC97-H, MHCC97-L, LM3, LM6) and a control cell line L-02. The expression profiles of Slit1, Slit2, Slit3, Robo1, and Robo3 were analyzed by reverse transcription polymerase chain reaction(RT-PCR). The methylation status of the promoters was detected by methylation specific polymerase chain reaction (MSP). Results The promoters of Slit1, Slit2 and Slit3 genes were almost

  11. Study on the methylation status of CDH1 and PAX1 genes in human cervical carcinoma tissues%人宫颈癌组织CDH1和PAX1基因甲基化研究

    Institute of Scientific and Technical Information of China (English)

    徐军; 王红琳; 陆杲川; 林晓

    2009-01-01

    目的:检测上皮型钙黏附素(E-cadherin,CDH1)基因和配对盒基因家族PAX1基因在宫颈癌组织及人乳头瘤病毒(human papillomavirus, HPV)感染的正常宫颈组织、宫颈炎组织、宫颈上皮内瘤样病变(cervical intraepithelial neoplasia, CIN)Ⅰ组织、CIN Ⅱ~Ⅲ组织和宫颈癌组织中的甲基化情况,评估是否可将其作为临床诊断的分子标志物.方法:应用甲基化特异性聚合酶链反应法(methylation specific PCR, MSP)对HPV感染的正常宫颈组织、宫颈炎组织、CINⅠ组织、CIN Ⅱ~Ⅲ组织以及宫颈癌组织中的CDH1和PAX1基因进行甲基化检测.结果:HPV感染的正常宫颈组织、宫颈炎组织和CINⅠ组织中未检出CDH1基因甲基化;CINⅡ~Ⅲ组织中检出CDH1基因甲基化2例(13.3%),宫颈癌组织中检出CDH1基因甲基化9例(22.5%),与HPV感染的正常宫颈组织比较差异均有统计学意义(P<0.05).HPV感染的正常宫颈组织和宫颈炎组织中未检出PAX1基因甲基化,CINⅠ、CINⅡ~Ⅲ和宫颈癌组织的PAX1基因甲基化阳性率分别为13.3%、46.7%和87.5%,CINⅡ~Ⅲ组织与CINⅠ和宫颈癌组织比较差异均有统计学意义(P<0.05).CINⅠ组织、CINⅡ~Ⅲ组织和宫颈癌组织的CDH1和PAX1基因甲基化总阳性率分别为13.3%、60.0%和97.5%.结论:CDH1和PAX1基因甲基化,尤其是PAX1基因甲基化对于宫颈癌临床诊断具有潜在价值.

  12. DNA methylation profiling of ovarian carcinomas and their in vitro models identifies HOXA9, HOXB5, SCGB3A1, and CRABP1 as novel targets

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    Tropé Claes G

    2007-07-01

    Full Text Available Abstract Background The epigenetics of ovarian carcinogenesis remains poorly described. We have in the present study investigated the promoter methylation status of 13 genes in primary ovarian carcinomas (n = 52 and their in vitro models (n = 4; ES-2, OV-90, OVCAR-3, and SKOV-3 by methylation-specific polymerase chain reaction (MSP. Direct bisulphite sequencing analysis was used to confirm the methylation status of individual genes. The MSP results were compared with clinico- pathological features. Results Eight out of the 13 genes were hypermethylated among the ovarian carcinomas, and altogether 40 of 52 tumours were methylated in one or more genes. Promoter hypermethylation of HOXA9, RASSF1A, APC, CDH13, HOXB5, SCGB3A1 (HIN-1, CRABP1, and MLH1 was found in 51% (26/51, 49% (23/47, 24% (12/51, 20% (10/51, 12% (6/52, 10% (5/52, 4% (2/48, and 2% (1/51 of the carcinomas, respectively, whereas ADAMTS1, MGMT, NR3C1, p14ARF, and p16INK4a were unmethylated in all samples. The methylation frequencies of HOXA9 and SCGB3A1 were higher among relatively early-stage carcinomas (FIGO I-II than among carcinomas of later stages (FIGO III-IV; P = 0.002, P = 0.020, respectively. The majority of the early-stage carcinomas were of the endometrioid histotype. Additionally, HOXA9 hypermethylation was more common in tumours from patients older than 60 years of age (15/21 than among those of younger age (11/30; P = 0.023. Finally, there was a significant difference in HOXA9 methylation frequency among the histological types (P = 0.007. Conclusion DNA hypermethylation of tumour suppressor genes seems to play an important role in ovarian carcinogenesis and HOXA9, HOXB5, SCGB3A1, and CRABP1 are identified as novel hypermethylated target genes in this tumour type.

  13. Identification of a Novel Methylated Gene in Nasopharyngeal Carcinoma: TTC40

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    Wajdi Ayadi

    2014-01-01

    Full Text Available To further explore the epigenetic changes in nasopharyngeal carcinoma (NPC, methylation-sensitive arbitrarily primed PCR was performed on NPC biopsies and nontumor nasopharyngeal samples. We have shown mainly two DNA fragments that appeared to be differentially methylated in NPCs versus nontumors. The first, defined as hypermethylated, corresponds to a CpG island at the 5′-end of the tetratricopeptide repeat domain 40 (TTC40 gene, whereas the second, defined as hypo-methylated, is located on repetitive sequences at chromosomes 16p11.1 and 13.1. Thereafter, the epigenetic alteration on the 5′-TTC40 gene was confirmed by methylation-specific PCR, showing a significant aberrant methylation in NPCs, compared to nontumors. In addition, the bisulfite sequencing analysis has shown a very high density of methylated cytosines in C15, C17, and X666 NPC xenografts. To assess whether TTC40 gene is silenced by aberrant methylation, we examined the gene expression by reverse transcription-PCR. Our analysis showed that the mRNA expression was significantly lower in tumors than in nontumors, which is associated with 5′-TTC40 gene hypermethylation. In conclusion, we found that the 5′-TTC40 gene is frequently methylated and is associated with the loss of mRNA expression in NPCs. Hypermethylation of 5′-TTC40 gene might play a role in NPC development; nevertheless, other studies are needed.

  14. SMARCB1/INI1-deficient sinonasal carcinoma shows methylation of RASSF1 gene: A clinicopathological, immunohistochemical and molecular genetic study of a recently described entity.

    Science.gov (United States)

    Laco, Jan; Chmelařová, Marcela; Vošmiková, Hana; Sieglová, Kateřina; Bubancová, Ivana; Dundr, Pavel; Němejcová, Kristýna; Michálek, Jaroslav; Čelakovský, Petr; Mottl, Radovan; Sirák, Igor; Vošmik, Milan; Ryška, Aleš

    2017-02-01

    The aim of the study was detailed clinicopathological investigation of SMARCB1/INI1-deficient sinonasal carcinomas, including molecular genetic analysis of mutational status and DNA methylation of selected protooncogenes and tumor suppressor genes by means of next generation sequencing (NGS) and methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA). A total of 4/56 (7%) cases of SMARCB1/INI1-deficient carcinomas were detected among 56 sinonasal carcinomas diagnosed over a 19year period using immunohistochemical screening. The series comprised 3 males and 1 female, aged 27-76 years (median 64 years). All tumors arose in the nasal cavity. Three neoplasms were diagnosed in advanced stage pT4. During the follow-up period (range 14-111 months (median 72 months)), three tumors recurred locally, but none of the patients developed regional or distant metastases. Ultimately, two patients died due to the tumor. Microscopically, all tumors consisted of infiltrating nests of polygonal basaloid cells with a variable component of rhabdoid cells with eosinophilic cytoplasm. Immunohistochemically, there was almost diffuse expression of cytokeratins (CK), p16, p40 and p63 in all cases, while expression of CK5/6, CK7 and vimentin was only focal or absent. The detection of NUT gave negative results. In three cases, the absence of SMARCB1/INI1 expression was due to deletion of SMARCB1/INI1 gene. Methylation of SMARCB1/INI1 gene was not found. One tumor harbored HPV18 E6/E7 mRNA. All 12 genes (BRAF, BRCA1, BRCA2, KIT, EGFR, KRAS, NRAS, PDGFRA, PIK3CA, PTEN, RET, and ROS1) tested for mutations using NGS were wild-type. Regarding DNA methylation, all four SMARCB1/INI1-deficient tumors showed methylation of RASSF1 gene by means of MS-MLPA. There was a statistically significant difference in RASSF1 gene methylation between SMARCB1/INI1-deficient and SMARCB1/INI1-positive tumors (p=0.0095). All other examined genes (ATM, BRCA1, BRCA2, CADM1, CASP8, CD44, CDKN1B

  15. Hepatocellular carcinoma displays distinct DNA methylation signatures with potential as clinical predictors.

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    Hector Hernandez-Vargas

    Full Text Available BACKGROUND: Hepatocellular carcinoma (HCC is characterized by late detection and fast progression, and it is believed that epigenetic disruption may be the cause of its molecular and clinicopathological heterogeneity. A better understanding of the global deregulation of methylation states and how they correlate with disease progression will aid in the design of strategies for earlier detection and better therapeutic decisions. METHODS AND FINDINGS: We characterized the changes in promoter methylation in a series of 30 HCC tumors and their respective surrounding tissue and identified methylation signatures associated with major risk factors and clinical correlates. A wide panel of cancer-related gene promoters was analyzed using Illumina bead array technology, and CpG sites were then selected according to their ability to classify clinicopathological parameters. An independent series of HCC tumors and matched surrounding tissue was used for validation of the signatures. We were able to develop and validate a signature of methylation in HCC. This signature distinguished HCC from surrounding tissue and from other tumor types, and was independent of risk factors. However, aberrant methylation of an independent subset of promoters was associated with tumor progression and etiological risk factors (HBV or HCV infection and alcohol consumption. Interestingly, distinct methylation of an independent panel of gene promoters was strongly correlated with survival after cancer therapy. CONCLUSION: Our study shows that HCC tumors exhibit specific DNA methylation signatures associated with major risk factors and tumor progression stage, with potential clinical applications in diagnosis and prognosis.

  16. Promoter methylation and expression of CDH1 and susceptibility and prognosis of eyelid squamous cell carcinoma.

    Science.gov (United States)

    Wang, Yong-Qiang; Yuan, Ye; Jiang, Shan; Jiang, Hua

    2016-07-01

    Eyelid skin tumors are the most frequent type of cancer in ophthalmology. And, eyelid squamous cell carcinoma (SCC) accounts for a large part of it. CDH1 encodes E-cadherin, a glycoprotein that plays an important part in cell-cell interaction. Loss of CDH1 function was suspected to be associated with tumorigenesis. Methylation of CDH1 promotors can alter the expression of its protein and is also considered as a contributor to various cancers. In this study, CDH1 methylation and expression profile as well as prognosis of 38 cases of eyelid SCC and the corresponding adjacent tissues were analyzed to clarify the role of CDH1 methylation in SCC carcinogenesis and prognosis. Methylation was detected by PCR, and CDH1 expression was evaluated by immunohistochemistry. We observed that CDH1 methylation is significantly correlated with decreased CDH1 protein expression in eyelid SCC patients. Patients with methylation and low expression of CDH1 are significantly associated with advanced and aggressive phenotypes. Therefore, CDH1 methylation and CDH1 expression are both independent prognostic factors for prognosis of eyelid SCC patients.

  17. Aberrant gene promoter methylation of p16, FHIT, CRBP1, WWOX, and DLC-1 in Epstein-Barr virus-associated gastric carcinomas.

    Science.gov (United States)

    He, Dan; Zhang, Yi-wang; Zhang, Na-na; Zhou, Lu; Chen, Jian-ning; Jiang, Ye; Shao, Chun-kui

    2015-04-01

    Alterations in global DNA methylation and specific regulatory gene methylation are frequently found in cancer, but the significance of these epigenetic changes in EBV-associated gastric carcinoma (EBVaGC) remains unclear. We evaluated global DNA methylation status in 49 EBVaGC and 45 EBV-negative gastric carcinoma (EBVnGC) tissue samples and cell lines by 5-methylcytosine immunohistochemical staining and methylation quantification. We determined promoter methylation status and protein expression for the p16, FHIT, CRBP1, WWOX, and DLC-1 genes in tissues and studied the correlation between CpG island methylator phenotype (CIMP) class and clinicopathological characteristics. Changes in gene methylation and mRNA expression in EBVaGC cell line SNU-719 and in EBVnGC cell lines SGC-7901, BGC-823, and AGS were assessed after treatment with 5-aza-2'-deoxycytidine (5-aza-dC), trichostatin A (TSA), or a combination of both, by methylation-specific PCR and quantitative real-time RT-PCR. Global genomic DNA hypomethylation was more pronounced in EBVnGC than in EBVaGC. Promoter methylation of all five genes was more frequent in EBVaGC than in EBVnGC (p < 0.05). p16 and FHIT methylation was reversely correlated with protein expression in EBVaGC. Most (41/49) EBVaGC exhibited CIMP-high (CIMP-H), and the prognosis of CIMP-H patients was significantly worse than that of CIMP-low (p = 0.027) and CIMP-none (p = 0.003) patients. Treatment with 5-aza-dC and/or TSA induced upregulation of RNA expression of all five genes in SNU-719; meanwhile, individual gene expression increased in EBVnGC cell lines. In summary, EBV-induced hypermethylation of p16, FHIT, CRBP1, WWOX, and DLC-1 may contribute to EBVaGC development. Demethylation therapy may represent a novel therapeutic strategy for EBVaGC.

  18. Methylation Status of H19/IGF2 Differentially Methylated Region in in vitro Human Blastocysts Donated by Healthy Couples

    Science.gov (United States)

    Derakhshan-Horeh, Marzieh; Abolhassani, Farid; Jafarpour, Farnoosh; Moini, Ashraf; Karbalaie, Khadijeh; Hosseini, Sayyed Morteza; Ostadhosseini, Somayyeh; Nasr-Esfahani, Mohammad Hossein

    2017-01-01

    Background: Imprinted genes are a unique subset of few genes that have been differentially methylated region (DMR) in a parental origin-dependent manner during gametogenesis, and these genes are highly protected during pre-implantation epigenetic reprogramming. Several studies have shown that the particular vulnerability of imprinting genes during suboptimal pre- and peri-conception micro-environments often is occurred by assisted reproduction techniques (ART). This study investigated the methylation status of H19/IGF2 DMR at high-quality expanding/expanded human blastocysts donated by healthy individuals to evaluate the risks linked to ART. Method: Methylation levels of H19/IGF2 DMR were analyzed by bisulfite conversion and sequencing at 18 CpG sites (CpGs) located in this region. Result: The overall percentage of methylated CpGs and the proportion of hyper-methylated clones of H19/IGF2 DMR in analyzed blastocysts were 37.85±4.87% and 43.75±5.1%, respectively. For validation of our technique, the corresponding methylation levels of peripheral human lymphocytes were defined (49.52±1.86% and 50%, respectively). Conclusion: Considering the absence of in vivo- produced human embryos, it is not possible to conclude that the methylation found in H19/IGF2 DMR is actually normal or abnormal. Regarding the possible risks associated with ART, the procedures should be optimized in order to at least reduce some of the epigenetic risks. PMID:27432596

  19. Methylation Status of Vitamin D Receptor Gene Promoter in Benign and Malignant Adrenal Tumors

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    Catia Pilon

    2015-01-01

    Full Text Available We previously showed a decreased expression of vitamin D receptor (VDR mRNA/protein in a small group of adrenocortical carcinoma (ACC tissues, suggesting the loss of a protective role of VDR against malignant cell growth in this cancer type. Downregulation of VDR gene expression may result from epigenetics events, that is, methylation of cytosine nucleotide of CpG islands in VDR gene promoter. We analyzed methylation of CpG sites in the VDR gene promoter in normal adrenals and adrenocortical tumor samples. Methylation of CpG-rich 5′ regions was assessed by bisulfite sequencing PCR using bisulfite-treated DNA from archival microdissected paraffin-embedded adrenocortical tissues. Three normal adrenals and 23 various adrenocortical tumor samples (15 adenomas and 8 carcinomas were studied. Methylation in the promoter region of VDR gene was found in 3/8 ACCs, while no VDR gene methylation was observed in normal adrenals and adrenocortical adenomas. VDR mRNA and protein levels were lower in ACCs than in benign tumors, and VDR immunostaining was weak or negative in ACCs, including all 3 methylated tissue samples. The association between VDR gene promoter methylation and reduced VDR gene expression is not a rare event in ACC, suggesting that VDR epigenetic inactivation may have a role in adrenocortical carcinogenesis.

  20. Correlation of MGMT promoter methylation status with gene and protein expression levels in glioblastoma

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    Miyuki Uno

    2011-01-01

    Full Text Available OBJECTIVES: 1 To correlate the methylation status of the O6-methylguanine-DNA-methyltransferase (MGMT promoter to its gene and protein expression levels in glioblastoma and 2 to determine the most reliable method for using MGMT to predict the response to adjuvant therapy in patients with glioblastoma. BACKGROUND: The MGMT gene is epigenetically silenced by promoter hypermethylation in gliomas, and this modification has emerged as a relevant predictor of therapeutic response. METHODS: Fifty-one cases of glioblastoma were analyzed for MGMT promoter methylation by methylation-specific PCR and pyrosequencing, gene expression by real time polymerase chain reaction, and protein expression by immunohistochemistry. RESULTS: MGMT promoter methylation was found in 43.1% of glioblastoma by methylation-specific PCR and 38.8% by pyrosequencing. A low level of MGMT gene expression was correlated with positive MGMT promoter methylation (p = 0.001. However, no correlation was found between promoter methylation and MGMT protein expression (p = 0.297. The mean survival time of glioblastoma patients submitted to adjuvant therapy was significantly higher among patients with MGMT promoter methylation (log rank = 0.025 by methylation-specific PCR and 0.004 by pyrosequencing, and methylation was an independent predictive factor that was associated with improved prognosis by multivariate analysis. DISCUSSION AND CONCLUSION: MGMT promoter methylation status was a more reliable predictor of susceptibility to adjuvant therapy and prognosis of glioblastoma than were MGMT protein or gene expression levels. Methylation-specific polymerase chain reaction and pyrosequencing methods were both sensitive methods for determining MGMT promoter methylation status using DNA extracted from frozen tissue.

  1. Comprehensive DNA Methylation Analysis Reveals a Common Ten-Gene Methylation Signature in Colorectal Adenomas and Carcinomas.

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    Árpád V Patai

    Full Text Available Microarray analysis of promoter hypermethylation provides insight into the role and extent of DNA methylation in the development of colorectal cancer (CRC and may be co-monitored with the appearance of driver mutations. Colonic biopsy samples were obtained endoscopically from 10 normal, 23 adenoma (17 low-grade (LGD and 6 high-grade dysplasia (HGD, and 8 ulcerative colitis (UC patients (4 active and 4 inactive. CRC samples were obtained from 24 patients (17 primary, 7 metastatic (MCRC, 7 of them with synchronous LGD. Field effects were analyzed in tissues 1 cm (n = 5 and 10 cm (n = 5 from the margin of CRC. Tissue materials were studied for DNA methylation status using a 96 gene panel and for KRAS and BRAF mutations. Expression levels were assayed using whole genomic mRNA arrays. SFRP1 was further examined by immunohistochemistry. HT29 cells were treated with 5-aza-2' deoxycytidine to analyze the reversal possibility of DNA methylation. More than 85% of tumor samples showed hypermethylation in 10 genes (SFRP1, SST, BNC1, MAL, SLIT2, SFRP2, SLIT3, ALDH1A3, TMEFF2, WIF1, whereas the frequency of examined mutations were below 25%. These genes distinguished precancerous and cancerous lesions from inflamed and healthy tissue. The mRNA alterations that might be caused by systematic methylation could be partly reversed by demethylation treatment. Systematic changes in methylation patterns were observed early in CRC carcinogenesis, occuring in precursor lesions and CRC. Thus we conclude that DNA hypermethylation is an early and systematic event in colorectal carcinogenesis, and it could be potentially reversed by systematic demethylation therapy, but it would need more in vitro and in vivo experiments to support this theory.

  2. Effects of Trophic Status on Mercury Methylation Pathways in Peatlands

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    Hines, M. E.; Zhang, L.; Sampath, S.; Hu, R.; Barkay, T.

    2014-12-01

    Methyl mercury (MeHg) is a bioaccumulative toxicant. It was believed to be produced by sulfate (SO4)- and iron- reducing bacteria (SRB and FeRB), but recent studies suggest that organisms that possess the gene cluster (hgcAB) can methylate Hg, which includes other microbial groups besides SRB and FeRB. Many areas known to accumulate high levels of MeHg are freshwater wetlands that lack sufficient electron acceptors to support the production of MeHg. To test the hypothesis that oligotrophic wetlands are able to methylate Hg by pathways that are not respiratory, peat was collected from three wetland sites in Alaska and three in Massachusetts that represented a trophic gradient. We determined rates of gas (CH4, CO2, H2) and LMW organic acid (formate, acetate, propionate, butyrate) formation, and rates of Hg methylation using the short-lived radioisotope 197Hg (half life 2.67 days). Two temperate sites exhibited strong terminal respiration (methanogenesis) and syntrophy, while the Alaskan sites and an oligotrophic temperate site exhibited low rates of both. Primary fermentation was an important process in the latter sites. Hg methylation was most active at the minerotrophic sites that exhibited active syntrophy and methanogenesis. Methylation decreased greatly in the presence of a methanogenic inhibitor and was stimulated by H2 indicating that methanogens and perhaps syntrophs were primary methylators. In the oligotrophic sites, the addition of SO4 stimulated methylation while a SO4 reduction inhibitor decreased methylation. There was no evidence of SO4 reduction in these samples suggesting that methylation was conducted by SRB that were metabolizing via fermentation and not SO4 reduction. While further studies are required to decipher the role of syntrophs including SRB varieties such as Syntrophobacter sp., these results indicate that fermentative bacteria may be able to significantly methylate Hg in wetlands that do not support anaerobic respiration.

  3. Mutation and methylation of hMLH1 in gastric carcinomas with microsatellite instability

    Institute of Scientific and Technical Information of China (English)

    Dian-Chun Fang; Rong-Quan Wang; Shi-Ming Yang; Jian-Ming Yang; Hai-Feng Liu; Gui-Yong Peng; Tian-Li Xiao; Yuan-Hui Luo

    2003-01-01

    AIM: To appraise the correlation of mutation and methylation of hMSH1 with microsatellite instability (MSI) in gastric cancers.METHODS: Mutation of hMLH1 was detected by Twodimensional electrophoresis (Two-D) and DNA sequencing;Methylation of hMLH1 promoter was measured with methylation-specific PCR; MST was analyzed by PCR-basecl methods.RESULTS: Sixty-eight cases of sporadic gastric carcinoma were studied for mutation and methylation of hMLH1 promoter and MSI. Three mutations were found, two of them were caused by a single bp substitution and one was caused by a 2 bp substitution, which displayed similar Two-D band pattern.Methylation of hMLH1 promoter was detected in 11(16.2%)gastric cancer. By using five MSI markers, MSI in at least one locus was detected in 17/68(25 %) of the tumors analyzed.Three hMLH1 mutations were all detected in MSI-H (≥2 loci,n=8), but no mutation was found in MSI-L (only one locus,n=9) or MSS (tumor lacking MSI or stable, n=51). Methylation frequency of hMLH1 in MSI-H (87.5 %, 7/8) was significantly higher than that in MSI-L (11.1%, 1/9) or MSS (5.9%, 3/51)(P<0.01-0.001), but no difference was found between MSI-L and MSS (P>0.05).CONCLUSION: Both mutation and methylation of hMLH1are involved in the MSI pathway but not related to the LOH pathway in gastric carcinogenesis.

  4. Trophic Status Controls Mercury Methylation Pathways in Northern Peats

    Science.gov (United States)

    Hines, M. E.; Zhang, L.; Barkay, T.; Krabbenhoft, D. P.; Schaefer, J.; Hu, H.; Sidelinger, W.; Liu, X.; Wang, Y.

    2015-12-01

    Methyl mercury (MeHg) can be produced by a variety of microbes including syntrophs, methanogens, acetogens, and fermenters, besides sulfate (SO42-, SRB) and iron- reducing bacteria. Many freshwater wetlands are deficient in electron acceptors that support the traditional respiratory pathways of methylation, yet they accumulate high levels of MeHg. To investigate methylation in these wetlands and to connect these pathways with vegetation and microbial communities, incubation experiments were conducted using peats from 26 sites in Alaska. The sites were clustered using multiple factor analysis based on pH, temp, CH4 and volatile fatty acids production rates, and surface vegetation composition. Three clusters were generated and corresponded to three trophic levels that were manifested by three pH levels (3.5, 4.5, and 5). Hg methylation activity in laboratory incubations was determined using the short-lived radioisotope 197Hg. In the low pH, Sphagnum-dominated cluster, methylation rates were less than 1% day-1 and likely conducted by primary fermenters. Conversely, the high pH trophic cluster dominated by Carex aquatilis and active syntrophy exhibited Hg methylation rates as high as 12% day-1. In intermediate sites, rich in Sphagnum magellanicum with less Carex, a gradient in syntrophy and Hg methylation paths was observed. Amendments with process-stimulators and inhibitors revealed no evidence of SO42- reduction, but suggested that SRB, metabolizing either syntrophically with methanogens and/or by fermentation, likely methylated Hg. While on going metatranscriptomics studies are required to verify the role of syntrophs, fermenters, and methanogens as methylators, these results revealed that Hg methylation pathways change greatly along trophic gradients with a dominance of respiratory pathways in mineral-rich sites, syntrophy dominance in intermediate sites, and fermentation dominance in nutrient-poor sites.

  5. Reduction of TIP30 in esophageal squamous cell carcinoma cells involves promoter methylation and microRNA-10b

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    Dong, Wenjie, E-mail: dongwenjie200581@126.com [Department of Internal Medicine-Oncology, The First Affiliated Hospital, Zhengzhou University (China); Shen, Ruizhe; Cheng, Shidan [Department of Gastroenterology, Rui-jin Hospital, Shanghai Jiao Tong University, Shanghai (China)

    2014-10-31

    Highlights: • TIP30 expression is frequently suppressed in ESCC. • TIP30 was hypermethylated in ESCC. • Reduction of TIP30 was significantly correlated with LN metastasis. • miR-10b is a direct regulator of TIP30. - Abstract: TIP30 is a putative tumor suppressor that can promote apoptosis and inhibit angiogenesis. However, the role of TIP30 in esophageal squamous cell carcinoma (ESCC) biology has not been investigated. Immunohistochemistry was used to investigate the expression of TIP30 in 70 ESCC. Hypermethylation of TIP30 was evaluated by the methylation specific PCR (MSP) method in ESCC (tumor and paired adjacent non-tumor tissues). Lost expression of TIP30 was observed in 50 of 70 (71.4%) ESCC. 61.4% (43 of 70) of primary tumors analyzed displayed TIP30 hypermethylation, indicating that this aberrant characteristic is common in ESCC. Moreover, a statistically significant inverse association was found between TIP30 methylation status and expression of the TIP30 protein in tumor tissues (p = 0.001). We also found that microRNA-10b (miR-10b) targets a homologous DNA region in the 3′untranslated region of the TIP30 gene and represses its expression at the transcriptional level. Reporter assay with 3′UTR of TIP30 cloned downstream of the luciferase gene showed reduced luciferase activity in the presence of miR-10b, providing strong evidence that miR-10b is a direct regulator of TIP30. These results suggest that TIP30 expression is regulated by promoter methylation and miR-10b in ESCC.

  6. Methylation matters? Decreased methylation status of genomic DNA in the blood of schizophrenic twins.

    Science.gov (United States)

    Bönsch, Dominikus; Wunschel, Michael; Lenz, Bernd; Janssen, Gesa; Weisbrod, Matthias; Sauer, Heinrich

    2012-08-15

    Studies of schizophrenia inheritance in identical twins show a concordance of about 50%, which supports an epigenetic model. In our present study we investigated methylation of genomic DNA and promoter methylation of Reelin and SOX10 genes in peripheral blood of twins suffering from schizophrenia. Global DNA methylation was reduced (52.3%) in schizophrenic twins if compared with healthy control twins (65.7%). The reduced methylation was significant in males only. We also found a similar hypomethylation in the non-affected twins of discordant pairs and a mixed group of psychiatric controls. In discordant twins there was a relative hypermethylation of the SOX10 promoter. Within-pair-difference of methylation of Reelin promoter was significantly lower in monozygotic twins than in dizygotic twins.

  7. Methylation Inactivation Mechanism of Parkin Gene mRNA Expression in Nasopharyngeal Carcinoma

    Institute of Scientific and Technical Information of China (English)

    Ni Haifeng; Jiang Bo; Zhou Zhen; Li Yong; Huang Guangwu

    2014-01-01

    Objective:To investigate the methylation inactivation and the clinical signiifcance of Parkin gene mRNA expression in nasopharyngeal carcinoma (NPC). Methods: The methylation-speciifc PCR (MSP) and semi-quantitative reverse transcription PCR (RT-PCR) were used to detect methylation and the mRNA expression level of Parkin gene in 54 cases of NPC tissues and 16 cases of normal nasopharyngeal epithelial (NNE) tissues.The mRNA expression of Parkin gene in two NPC cell lines (CNE1 and CNE2) were detected before and after treatment with the methyltransferase inhibitor 5-aza-2-deoxycytidine so as to analyze the effects of Parkin gene methylation and demethylation on Parkin gene mRNA expression and the relationship between Parkin gene mRNA expression and clinical factors. Results: The methylation frequency of Parkin gene in human NPC tissues was 62.96% (34/54), but didn’t happen in any of 16 cases of NNE tissues. The mRNA expression level was (0.3430±0.4947) in 54 cases of NPC tissues and (1.0052±0.4911) in NNE tissues, showing that the mRNA expression level of NPC tissues was significantly down-regulated (P0.05), but was closely related to lymph node metastasis (P<0.05). Conclusion:Parkin gene mRNA expression, serving as a cancer suppressor gene in the occurrence and development of NPC, is inactivated and regulated by methylation, which also has a negative correlation with lymph node metastasis and could be considered as the judgment of predictive index of clinical prognosis of NPC.

  8. High-definition DNA methylation profiles from breast and ovarian carcinoma cell lines with differing doxorubicin resistance.

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    Michael Boettcher

    Full Text Available Acquired drug resistance represents a frequent obstacle which hampers efficient chemotherapy of cancers. The contribution of aberrant DNA methylation to the development of drug resistant tumor cells has gained increasing attention over the past decades. Hence, the objective of the presented study was to characterize DNA methylation changes which arise from treatment of tumor cells with the chemotherapeutic drug doxorubicin. DNA methylation levels from CpG islands (CGIs linked to twenty-eight genes, whose expression levels had previously been shown to contribute to resistance against DNA double strand break inducing drugs or tumor progression in different cancer types were analyzed. High-definition DNA methylation profiles which consisted of methylation levels from 800 CpG sites mapping to CGIs around the transcription start sites of the selected genes were determined. In order to investigate the influence of CGI methylation on the expression of associated genes, their mRNA levels were investigated via qRT-PCR. It was shown that the employed method is suitable for providing highly accurate methylation profiles, comparable to those obtained via clone sequencing, the gold standard for high-definition DNA methylation studies. In breast carcinoma cells with acquired resistance against the double strand break inducing drug doxorubicin, changes in methylation of specific cytosines from CGIs linked to thirteen genes were detected. Moreover, similarities between methylation profiles obtained from breast and ovarian carcinoma cell lines with acquired doxorubicin resistance were found. The expression levels of a subset of analyzed genes were shown to be linked to the methylation levels of the analyzed CGIs. Our results provide detailed DNA methylation information from two separate model systems for acquired doxorubicin resistance and suggest the occurrence of similar methylation changes in both systems upon exposure to the drug.

  9. Distinctive Klf4 mutants determine preference for DNA methylation status

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    Hashimoto, Hideharu; Wang, Dongxue; Steves, Alyse N.; Jin, Peng; Blumenthal, Robert M.; Zhang, Xing; Cheng, Xiaodong

    2016-09-04

    Reprogramming of mammalian genome methylation is critically important but poorly understood. Klf4, a transcription factor directing reprogramming, contains a DNA binding domain with three consecutive C2H2 zinc fingers. Klf4 recognizes CpG or TpG within a specific sequence. Mouse Klf4 DNA binding domain has roughly equal affinity for methylated CpG or TpG, and slightly lower affinity for unmodified CpG. The structural basis for this key preference is unclear, though the side chain of Glu446 is known to contact the methyl group of 5-methylcytosine (5mC) or thymine (5-methyluracil). We examined the role of Glu446 by mutagenesis. Substituting Glu446 with aspartate (E446D) resulted in preference for unmodified cytosine, due to decreased affinity for 5mC. In contrast, substituting Glu446 with proline (E446P) increased affinity for 5mC by two orders of magnitude. Structural analysis revealed hydrophobic interaction between the proline's aliphatic cyclic structure and the 5-methyl group of the pyrimidine (5mC or T). As in wild-type Klf4 (E446), the proline at position 446 does not interact directly with either the 5mC N4 nitrogen or the thymine O4 oxygen. In contrast, the unmethylated cytosine's exocyclic N4 amino group (NH2) and its ring carbon C5 atom hydrogen bond directly with the aspartate carboxylate of the E446D variant. Both of these interactions would provide a preference for cytosine over thymine, and the latter one could explain the E446D preference for unmethylated cytosine. Finally, we evaluated the ability of these Klf4 mutants to regulate transcription of methylated and unmethylated promoters in a luciferase reporter assay.

  10. PRSS8 methylation and its significance in esophageal squamous cell carcinoma

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    Bao, Yonghua; Wang, Qian; Guo, Yongchen; Chen, Zhiguo; Li, Kai; Yang, Yiqiong; Zhang, Huijuan; Dong, Huali; Shen, Kui; Yang, Wancai

    2016-01-01

    Esophageal cancer is one of the most common cancers worldwide, and the incidence and mortality is increasing rapidly in recent years in China, but the underlying mechanisms are largely unclear. Herein we found that the expression of PRSS8, a serine protease prostasin, is significantly decreased in esophageal squamous cell carcinomas (ESCC) at mRNA and protein levels. The reduction of PRSS8 was well correlated with poor differentiation and shorter survival time. Interestingly, ESCC stromal expression of PRSS8 was significantly correlated with stromal lymphocyte infiltration and cancer progression. Methylation specific PCR showed that PRSS8 was hypermethylated in ESCC tissues and ESCC cell lines, which was linked to the downregulation of PRSS8 expression and decreased activities of PRSS8 promoter. De-methylation agent decitabine was able to restore PRSS8 expression, leading to the inhibition of cancer cell proliferation, motility, migration and cell cycle arrest. However, the restored PRSS8 and its tumor inhibition could be reversed by small interfering RNA targeting PRSS8. Mechanistic study showed that tumor inhibition of PRSS8 may be associated with proliferation- and epithelial mesenchymal transition - related proteins in ESCC cells. In conclusion, our finding showed that PRSS8 methylation and its stromal expression had important clinical significance in ESCC. PMID:27081034

  11. Highly efficient generation of transgenic sheep by lentivirus accompanying the alteration of methylation status.

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    Chenxi Liu

    Full Text Available BACKGROUND: Low efficiency of gene transfer and silence of transgene expression are the critical factors hampering the development of transgenic livestock. Recently, transfer of recombinant lentivirus has been demonstrated to be an efficient transgene delivery method in various animals. However, the lentiviral transgenesis and the methylation status of transgene in sheep have not been well addressed. METHODOLOGY/PRINCIPLE FINDINGS: EGFP transgenic sheep were generated by injecting recombinant lentivirus into zygotes. Of the 13 lambs born, 8 carried the EGFP transgene, and its chromosomal integration was identified in all tested tissues. Western blotting showed that GFP was expressed in all transgenic founders and their various tissues. Analysis of CpG methylation status of CMV promoter by bisulfate sequencing unraveled remarkable variation of methylation levels in transgenic sheep. The average methylation levels ranged from 37.6% to 79.1% in the transgenic individuals and 34.7% to 83% in the tested tissues. Correlative analysis of methylation status with GFP expression revealed that the GFP expression level was inversely correlated with methylation density. The similar phenomenon was also observed in tested tissues. Transgene integration determined by Southern blotting presented multiple integrants ranging from 2 to 6 copies in the genome of transgenic sheep. CONCLUSIONS/SIGNIFICANCE: Injection of lentiviral transgene into zygotes could be a promising efficient gene delivery system to generate transgenic sheep and achieved widespread transgene expression. The promoter of integrants transferred by lentiviral vector was subjected to dramatic alteration of methylation status and the transgene expression level was inversely correlative with promoter methylation density. Our work illustrated for the first time that generation of transgenic sheep by injecting recombinant lentivirus into zygote could be an efficient tool to improve sheep performance by

  12. Protein expression and methylation of MGMT, a DNA repair gene and their correlation with clinicopathological parameters in invasive ductal carcinoma of the breast.

    Science.gov (United States)

    Asiaf, Asia; Ahmad, Shiekh Tanveer; Malik, Ajaz Ahmad; Aziz, Shiekh Aejaz; Rasool, Zubaida; Masood, Akbar; Zargar, Mohammad Afzal

    2015-08-01

    Epigenetic mechanisms such as DNA methylation are being increasingly recognized to play an important role in cancer and may serve as a cancer biomarker. The aim of this study was to evaluate the promoter methylation status of MGMT (O6-methylguanine-DNA methyltransferase) and a possible correlation with the expression of MGMT and standard clinicopathological parameters in invasive ductal breast carcinoma patients (IDC) of Kashmir. Methylation-specific PCR was carried out to investigate the promoter methylation status of MGMT in breast tumors paired with the corresponding normal tissue samples from 128 breast cancer patients. The effect of promoter methylation on protein expression in the primary breast cancer and adjacent normal tissues was evaluated by immunohistochemistry (n = 128) and western blotting (n = 30). The frequency of tumor hypermethylation was 39.8 % and a significant difference in methylation frequency among breast tumors were found (p MGMT in 68/128 (53.1 %) tumors. MGMT promoter methylation mediated gene silencing was associated with loss of its protein expression (rs = -0.285, p = 0.001, OR = 3.38, 95 % CI = 1.59-7.17). A significant correlation was seen between loss of MGMT and lymph node involvement (p = 0.030), tumor grade (p MGMT methylation was found to be associated with tumor grade (p = 0.011), tumor stage (p = 0.009), and loss of ER (p = 0.003) and PR receptors (p = 0.009). To our knowledge, our findings, for the first time, in Kashmiri population, indicate that MGMT is aberrantly methylated in breast cancer and promoter hypermethylation could be attributed to silencing of MGMT gene expression in breast cancer. Our data suggests that MGMT promoter hypermethylation could have a potential function as molecular biomarker of breast oncogenesis. Also, based on their predictive value of response to therapy, the immunohistochemical evaluation and interpretation of MGMT may also help in future to

  13. Downregulation of retinoic acid receptor-β2expression is linked to aberrant methylation in esophageal squamous cell carcinoma cell lines

    Institute of Scientific and Technical Information of China (English)

    Zhong-Min Liu; Fang Ding; Ming-Zhou Guo; Li-Yong Zhang; Min Wu; Zhi-Hua Liu

    2004-01-01

    AIM: To study the role of hypermethylation in the loss ofretinoic acid receptorβ2(RARβ2) in esophageal squamous cell carcinoma (ESCC).METHODS: The role of hypermethylation in RAR,β2 gene silencing in 6 ESCC cell lines was determined by methylationspecific PCR (MSP), and its methylation status was compared with RARβ2 mRNA expression by RT-PCR. The MSP results were confirmed by bisulfite sequencing of RARβ2promoter regions. RESULTS: Methylation was detected in 4 of the 6 cell lines, and the expression of RARβ2was markedly downregulated in 3 of the 4 methylated cell lines. The expression of RARβ2was restored in one RARβ2-downregulated cell line with the partial demethylation of promoter region of RARβ after 5aza-2'-deoxycytidine (5-aza-dc) treatment.CONCLUSION: The methylation of the 5' region may play an important role in the downregulation of RARβ2 in someESCC cell lines, suggesting that multiple mechanisms contribute to the loss of RARβ2expression in ESCC cell lines. This study may have clinical applications for treatment and prevention of ESCC.

  14. Methylation status of the interferon-gamma gene promoter in chronic hepatitis B

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Objective To evaluate the methylation status at CpG site -55 in the interferon-gamma (IFN-γ) gene promoter and its effect on IFN-γ expression in chronic hepatitis B. Method The authors recruited 30 patients with HBeAg-positive chronic hepatitis B (CHB), 30 HBeAg-negative CHB patients, and 30 healthy blood donors. Pyrosequencing was used to determine the methylation status at CpG site -55 in the IFN-γ gene promoter following bisulfite treatment of DNA in peripheral blood mononuclear cells (PBMCs). The expres...

  15. Nucleosome Positions and Differential Methylation Status of Various Regions within MLH1 CpG Island

    Institute of Scientific and Technical Information of China (English)

    BAI Hua; ZHOU Jing; DENG Da-jun

    2008-01-01

    Objective:To determine the relationship between nucleosome positions and formation of differential methylation of the reported region A,B,C,and D within the MLH1 CpG island. Methods:Methylation of the MLH1 promoter was analyzed by combined of bisulfite restriction assay.Chromatin of RKO and MGC803 cells were extracted and digested by Mnase.Mononucleosomal DNA fragment was isolated and used as templates for detection of nucleosomal distribution by a battery of quantitative PCRs covering the full MLH1 promoter region. Results:The MLH1 was methylated in RKO and unmethylated in MGC803.At the region B,where methylation of CpG sites did not correlated with transcription of this gene well,qPCR product of the M-3(-599nt~-475nt)fragment was amplified in both RKO and MGC803 cells.However,at the region C and D within the core promoter,where methylation of CpG sites correlated with loss of MLH1 transcription well,the M-7(-257nt~-153nt)and M-8(-189nt~-71nt)fragments were amplified remarkably only in RKO cells. Conclusion:Nucleosome may be the basic unit for both CpG methylation and methylation-related regulation of gene transcription.Methylation status of CpG sites within the same nucleosome may be homogeneous;between different nucleosomes,homogeneous or heterogeneous.

  16. Characterisation of DNA methylation status using spectroscopy (mid-IR versus Raman) with multivariate analysis.

    Science.gov (United States)

    Kelly, Jemma G; Najand, Ghazal M; Martin, Francis L

    2011-05-01

    Methylation status plays important roles in the regulation of gene expression and significantly influences the dynamics, bending and flexibility of DNA. The aim of this study was to determine whether attenuated total reflection Fourier-transform infrared (ATR-FTIR) or Raman spectroscopy with subsequent multivariate analysis could determine methylation patterning in oligonucleotides variously containing 5-methylcytosine, cytosine and guanine bases. Applied to Low-E reflective glass slides, 10 independent spectral acquisitions were acquired per oligonucleotide sample. Resultant spectra were baseline-corrected and vector normalised over the 1750 cm(-1) -760 cm(-1) (for ATR-FTIR spectroscopy) or the 1750 cm(-1) -600 cm(-1) (for Raman spectroscopy) regions. Data were then analysed using principal component analysis (PCA) coupled with linear discriminant analysis (LDA). Exploiting this approach, biomolecular signatures enabling sensitive and specific discrimination of methylation patterning were derived. For DNA sequence and methylation analysis, this approach has the potential to be an important tool, especially when material is scarce.

  17. Methylation and protein expression of DNA repair genes: association with chemotherapy exposure and survival in sporadic ovarian and peritoneal carcinomas

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    Walsh Tom

    2009-07-01

    Full Text Available Abstract Background DNA repair genes critically regulate the cellular response to chemotherapy and epigenetic regulation of these genes may be influenced by chemotherapy exposure. Restoration of BRCA1 and BRCA2 mediates resistance to platinum chemotherapy in recurrent BRCA1 and BRCA2 mutated hereditary ovarian carcinomas. We evaluated BRCA1, BRCA2, and MLH1 protein expression in 115 sporadic primary ovarian carcinomas, of which 31 had paired recurrent neoplasms collected after chemotherapy. Additionally, we assessed whether promoter methylation of BRCA1, MLH1 or FANCF influenced response to chemotherapy or explained alterations in protein expression after chemotherapy exposure. Results Of 115 primary sporadic ovarian carcinomas, 39 (34% had low BRCA1 protein and 49 (42% had low BRCA2 expression. BRCA1 and BRCA2 protein expression were highly concordant (p Conclusion Low BRCA1 expression in primary sporadic ovarian carcinoma is associated with prolonged survival. Recurrent ovarian carcinomas commonly have increased BRCA1 and/or BRCA2 protein expression post chemotherapy exposure which could mediate resistance to platinum based therapies. However, alterations in expression of these proteins after chemotherapy are not commonly mediated by promoter methylation, and other regulatory mechanisms are likely to contribute to these alterations.

  18. Screening of candidate tumor-suppressor genes in 3p21.3 and investigation of the methylation of gene promoters in oral squamous cell carcinoma.

    Science.gov (United States)

    Wang, Kai; Ling, Tianyou; Wu, Hanjiang; Zhang, Jie

    2013-03-01

    Oral squamous cell carcinoma (OSCC) is the most common type of head and neck malignant tumor. however, its pathological mechanisms have not yet been elucidated. In the present study, we screened for candidate tumor-suppressor genes (TSGs) related to OSCC among 10 candidate genes located in 3p21.3, a region abundant with TSGs based on previous studies, using semi-quantitative reverse transcription PCR (RT-PCR). Three genes, GNAT1, SEMA3B and AXUD1, with low or no expression in OSCC tissues and the cell line TCA8113 were selected, and the promoter methylation status was further analyzed by methylation-specific PCR (MS-PCR). Hypermethylation in the promoter regions of SEMA3B was found in OSCC tissues, and a significant difference in the frequency of methylation of SEMA3B was observed between OSCC and non-cancerous tissues. Furthermore, TCA8113 cells treated with 5-Aza-Cdc started to re-express SEMA3B at a concentration of 5 µM or higher. Our study confirmed that three candidate TSGs with low expression may be involved in OSCC and that hypermethylation in promoter regions may contribute to the low expression of SEMA3B. These findings offer novel insights for clarifying the molecular mechanisms of tumorigenesis of OSCC as well as for aiding in its clinical diagnosis and therapeutic strategy.

  19. Methylation status of the interferon-gamma gene promoter in chronic hepatitis B

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    Objective To evaluate the methylation status at CpG site -55 in the interferon-gamma (IFN-7) gene promoter and its effect on IFN-7 expression in chronic hepatitis B. Method The authors recruited 30 patients with UBeAg-positive chronic hepatitis B (CHB), 30 HBeAg-negative CHB patients, and 30 healthy blood donors. Pyrosequeneing was used to determine the methylation status at CpG site -55 in the IFN-γ gene promoter following bisulfite treatment of DNA in peripheral blood mononuclear cells (PBMCs). The expression of IFN-γ was analyzed by real-time RT-PCR and ELISA. HBV DNA in PBMCs was detected by nested PCR. Results The methylation level at CpG site -55 in the IFN-γ gene promoter was significantly increased, resulting in subsequent down-regulation of the expression of this cytoldne in CHB. The methylation level at CpG site -55 was significantly higher in HBeAg-positive patients than in HBeAg-negative ones (P<0.01) and was also significantly higher in PBMCs from HBV DNA-positive patients than from HBV DNA-negative ones (P<0.01) ; the methylation level at CpG site -55 was positively correlated with the amount of HBV DNA in serum (P<0.01). Oonclusion IFN-γ gene expression appears to be regulated by methylation of the IFN-γ gene promoter in CHB; the methylation level at CpG site -55 is associated with HBV infection.

  20. Methylation status of c-fms oncogene in HCC and its relationship with clinical pathology

    Institute of Scientific and Technical Information of China (English)

    Jun Cui; Dong Hua Yang; Xiang Jun Bi; Zi Rong Fan

    2001-01-01

    @@ INTRODUCTIONThe mechanism that DNA hypomethylation leads toactivation of oncogene and occurrence of malignantneoplasm is being increasingly recognized byresearchers. Normal DNA methylation playsimportant role in stabilizing the phenotype of cell.DNA methylation status reduction and/or patternalteration are related to activation and abnormallyhigh expression of some oncogenes and cellularmalignancy[1-6]. c-fms oncogene encodes for colonystimulating factor 1 receptor (CSF-1R)[7], c-fms/CSF-1R was highly expressed in hepatocellularcarcinoma (HCC) tissue, but the mechanismremained obscure[8,9].

  1. DGKI methylation status modulates the prognostic value of MGMT in glioblastoma patients treated with combined radio-chemotherapy with temozolomide.

    Directory of Open Access Journals (Sweden)

    Amandine Etcheverry

    Full Text Available BACKGROUND: Consistently reported prognostic factors for glioblastoma (GBM are age, extent of surgery, performance status, IDH1 mutational status, and MGMT promoter methylation status. We aimed to integrate biological and clinical prognostic factors into a nomogram intended to predict the survival time of an individual GBM patient treated with a standard regimen. In a previous study we showed that the methylation status of the DGKI promoter identified patients with MGMT-methylated tumors that responded poorly to the standard regimen. We further evaluated the potential prognostic value of DGKI methylation status. METHODS: 399 patients with newly diagnosed GBM and treated with a standard regimen were retrospectively included in this study. Survival modelling was performed on two patient populations: intention-to-treat population of all included patients (population 1 and MGMT-methylated patients (population 2. Cox proportional hazard models were fitted to identify the main prognostic factors. A nomogram was developed for population 1. The prognostic value of DGKI promoter methylation status was evaluated on population 1 and population 2. RESULTS: The nomogram-based stratification of the cohort identified two risk groups (high/low with significantly different median survival. We validated the prognostic value of DGKI methylation status for MGMT-methylated patients. We also demonstrated that the DGKI methylation status identified 22% of poorly responding patients in the low-risk group defined by the nomogram. CONCLUSIONS: Our results improve the conventional MGMT stratification of GBM patients receiving standard treatment. These results could help the interpretation of published or ongoing clinical trial outcomes and refine patient recruitment in the future.

  2. Keap1/Nrf2 pathway in kidney cancer: frequent methylation of KEAP1 gene promoter in clear renal cell carcinoma.

    Science.gov (United States)

    Fabrizio, Federico Pio; Costantini, Manuela; Copetti, Massimiliano; la Torre, Annamaria; Sparaneo, Angelo; Fontana, Andrea; Poeta, Luana; Gallucci, Michele; Sentinelli, Steno; Graziano, Paolo; Parente, Paola; Pompeo, Vincenzo; De Salvo, Laura; Simone, Giuseppe; Papalia, Rocco; Picardo, Francesco; Balsamo, Teresa; Flammia, Gerardo Paolo; Trombetta, Domenico; Pantalone, Angela; Kok, Klaas; Paranita, Ferronika; Muscarella, Lucia Anna; Fazio, Vito Michele

    2017-01-04

    The Keap1/Nrf2 pathway is a master regulator of the cellular redox state through the induction of several antioxidant defence genes implicated in chemotherapeutic drugs resistance of tumor cells. An increasing body of evidence supports a key role for Keap1/Nrf2 pathway in kidney diseases and renal cell carcinoma (RCC), but data concerning the molecular basis and the clinical effect of its deregulation remain incomplete.Here we present a molecular profiling of the KEAP1 and NFE2L2 genes in five different Renal Cell Carcinoma histotypes by analysing 89 tumor/normal paired tissues (clear cell Renal Carcinoma, ccRCCs; Oncocytomas; Papillary Renal Cell Carcinoma Type 1, PRCC1; Papillary Renal Cell Carcinoma Type 2, PRCC2; and Chromophobe Cell Carcinoma).A tumor-specific DNA methylation of the KEAP1 gene promoter region was found as a specific feature of the ccRCC subtype (18/37, 48.6%) and a direct correlation with mRNA levels was confirmed by in vitro 5-azacytidine treatment. Analysis of an independent data set of 481 ccRCC and 265 PRCC tumors corroborates our results and multivariate analysis reveals a significant correlation among ccRCCs epigenetic KEAP1 silencing and staging, grading and overall survival.Our molecular results show for the the first time the epigenetic silencing of KEAP1 promoter as the leading mechanism for modulation of KEAP1 expression in ccRCCs and corroborate the driver role of Keap1/Nrf2 axis deregulation with potential new function as independent epigenetic prognostic marker in renal cell carcinoma.

  3. LOH at chromosome 9q34.3 and the Notch1 gene methylation are less involved in oral squamous cell carcinomas

    DEFF Research Database (Denmark)

    Gao, Shan; Krogdahl, Annelise; Eiberg, Hans;

    2007-01-01

    BACKGROUND: Previous studies of oral carcinomas have shown that both loss of heterozygosity (LOH) and hypermethylation at chromosome 9q33 to 9q34.2 are frequent. The present study investigates the frequency of Notch1 gene methylation and LOH at 9q34.3 region. METHODS: Gene promoter hypermethylation...... of the Notch1 gene was analysed by methylation-specific PCR and LOH was analysed using microsatellite markers. RESULTS: We found LOH at 9q34.3 in three patients and methylation of the Notch1 gene only in two patients with oral carcinoma. CONCLUSION: Comparing with the alterations at 9q33 to 34.2 regions, LOH...... at 9q34.3 and methylation of the Notch1 gene was less involved in oral squamous cell carcinomas....

  4. Liver insulin-like growth factor 2 methylation in hepatitis C virus drrhosis and further occurrence of hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Philippe Couvert; Michel Beaugrand; Nathalie Ganne-Carrié; Alain Carrié; Jacques Pariès; Jenny Vaysse; Audrey Miroglio; Antoine Kerjean; Pierre Nahon; Jamel Chelly; Jean-Claude Trinchet

    2008-01-01

    AIM: To assess the predictive value of the insulinlike growth factor 2 (Igf2) methylation profile for the occurrence of Hepatocellular Carcinoma (HCC) in hepatitis C (HCV) cirrhosis.METHODS: Patients with: (I) biopsy-proven compensated HCV cirrhosis; (2) available baseline frozen liver sample; (3) absence of detectable HCC; (4) regular screening for HCC; (5) informed consent for genetic analysis were studied.After DNA extraction from liver samples and bisulfite treatment,unbiased PCR and DHPLC analysis were performed for methylation analysis at the Igf2 locus.The predictive value of the Igf2 methylation profile for HCC was assessed by Kaplan-Meier and Cox methods.RESULTS: Among 94 included patients,20 developed an HCC during follow-up (6.9±3.2 years).The methylation profile was hypomethylated,intermediate and hypermethylated in 13,64 and 17 cases,respectively.In univariate analysis,two baseline parameters were associated with the occurrence of HCC: age (P=0.01) and prothrombin (P=0.04).The test of linear tendency between the three ordered levels of Igf2 methylation and probability of HCC occurrence was significant (Log Rank,P=0.043;Breslow,P=0.037; Tarone-Ware,P=0.039).CONCLUSION: These results suggest that hypomethylation at the Igf2 locus in the liver could be predictive for HCC occurrence in HCV cirrhosis.

  5. Effects of methylation status of caspase-8 promoter on antitumor activity of TRAIL to human gastric cancer cells

    Institute of Scientific and Technical Information of China (English)

    ZHANG Ru-gang; FANG Dian-chun; YANG Liu-qin; LUO Yuan-gang

    2004-01-01

    Objective: To study the effects of the methylation status of caspase-8 promoter on the antitumor activity of TRAIL to the human gastric cancer cells. Methods: The methylation of caspase-8 was measured with methylation specific PCR (MSP) and the antitomor capability of TRAIL to human gastric cancer cells was determined with MTT. Results: No methylation of caspase-8 in the human gastric cancer cells was found. The sensitivity of 5 lines of gastric cancer cells to the antitumor activity of TRAIL was different. The administration of the demethylation agent 5-Aza-2'-deoxycytidine ( 5-AzaCdR) increased the sensitivity of gastric cancer cells to TRAIL but did not change the methylation status of caspase-8 promoter in gastric cancer cells. Conclusion: 5-Aza-CdR increases the sensitivity of most of gastric cancer cells to TRAIL but caspase-8 is not involved in the antitumor activity of TRAIL.

  6. Aberrant methylation of the adenomatous polyposis coli (APC) gene promoter 1A in breast and lung carcinomas.

    Science.gov (United States)

    Virmani, A K; Rathi, A; Sathyanarayana, U G; Padar, A; Huang, C X; Cunnigham, H T; Farinas, A J; Milchgrub, S; Euhus, D M; Gilcrease, M; Herman, J; Minna, J D; Gazdar, A F

    2001-07-01

    The adenomatous polyposis coli (APC) gene is a tumor suppressor gene associated with both familial and sporadic cancer. Despite high rates of allelic loss in lung and breast cancers, point mutations of the APC gene are infrequent in these cancer types. Aberrant methylation of the APC promoter 1A occurs in some colorectal and gastric malignancies, and we investigated whether the same mechanism occurs in lung and breast cancers. The methylation status of the APC gene promoter 1A was analyzed in 77 breast, 50 small cell (SCLC), and 106 non-small cell (NSCLC) lung cancer tumors and cell lines and in 68 nonmalignant tissues by methylation-specific PCR. Expression of the APC promoter 1A transcript was examined in a subset of cell lines by reverse transcription-PCR, and loss of heterozygosity at the gene locus was analyzed by the use of 12 microsatellite and polymorphic markers. Statistical tests were two-sided. Promoter 1A was methylated in 34 of 77 breast cancer tumors and cell lines (44%), in 56 of 106 NSCLC tumors and cell lines (53%), in 13 of 50 SCLC cell lines (26%), and in 3 of 68 nonmalignant samples (4%). Most cell lines tested contained the unmethylated or methylated form exclusively. In 27 cell lines tested, there was complete concordance between promoter methylation and silencing of its transcript. Demethylation with 5-aza-2'-deoxycytidine treatment restored transcript 1A expression in all eight methylated cell lines tested. Loss of heterozygosity at the APC locus was observed in 85% of SCLCs, 83% of NSCLCs, and 63% of breast cancer cell lines. The frequency of methylation in breast cancers increased with tumor stage and size. In summary, aberrant methylation of the 1A promoter of the APC gene and loss of its specific transcript is frequently present in breast and NSCLC cancers and cell lines and, to a lesser extent, in SCLC cell lines. Our findings may be of biological and clinical importance.

  7. Immunotherapy for Urothelial Carcinoma: Current Status and Perspectives

    Directory of Open Access Journals (Sweden)

    Taiji Tsukamoto

    2011-07-01

    Full Text Available Intravesical instillation of bacillus Calmette Guérin (BCG for the treatment of urothelial carcinoma (UC of the bladder is based on the BCG-induced immune response, which eradicates and prevents bladder cancer. The results of recent studies have suggested that not only major histocompatibility complex (MHC-nonrestricted immune cells such as natural killer cells, macrophages, neutrophils, etc., but also MHC-restricted CD8+ T cells play an important role and are one of the main effectors in this therapy. Better understanding of the mechanism of BCG immunotherapy supports the idea that active immunotherapy through its augmented T cell response can have great potential for the treatment of advanced UC. In this review, progress in immunotherapy for UC is discussed based on data from basic, translational and clinical studies. We also review the escape mechanism of cancer cells from the immune system, and down-regulation of MHC class I molecules.

  8. Immunotherapy for Urothelial Carcinoma: Current Status and Perspectives

    Energy Technology Data Exchange (ETDEWEB)

    Kitamura, Hiroshi, E-mail: hkitamu@sapmed.ac.jp; Tsukamoto, Taiji [Department of Urology, Sapporo Medical University School of Medicine, South 1 West 16, Chuo-ku, Sapporo 060-8543 (Japan)

    2011-07-29

    Intravesical instillation of bacillus Calmette Guérin (BCG) for the treatment of urothelial carcinoma (UC) of the bladder is based on the BCG-induced immune response, which eradicates and prevents bladder cancer. The results of recent studies have suggested that not only major histocompatibility complex (MHC)-nonrestricted immune cells such as natural killer cells, macrophages, neutrophils, etc., but also MHC-restricted CD8{sup +} T cells play an important role and are one of the main effectors in this therapy. Better understanding of the mechanism of BCG immunotherapy supports the idea that active immunotherapy through its augmented T cell response can have great potential for the treatment of advanced UC. In this review, progress in immunotherapy for UC is discussed based on data from basic, translational and clinical studies. We also review the escape mechanism of cancer cells from the immune system, and down-regulation of MHC class I molecules.

  9. MOLECULAR GENETIC DISORDERS IN THE VHL GENE AND METHYLATION OF SOME SUPPRESSOR GENES IN SPORADIC CLEAR-CELL RENAL CARCINOMAS

    Directory of Open Access Journals (Sweden)

    D. S. Mikhailenko

    2014-07-01

    Full Text Available Renal carcinoma (RC is one of ten most common malignancies in adults and an urgent problem of modern oncology. The purpose of the study was to make a molecular genetic analysis of a number of suppressor genes in RC, which was aimed at searching for and characterizing the potential markers of the disease. Two hundred and nine RC samples were examined, of them there were 192 clear-cell carcinomas. VHL gene mutations were detected by single-strand conformation polymorphism and sequence analyses while the methylation of suppressor genes was by the methylation-sensitive polymerase chain reaction. Somatic VHL mutations were determined in 35.4% of cases of clear-cell RC (CCRC. VHL gene disorders were found in 53.7% of patients with Stage 1, which counts in favor of VHL inactivation in early-stage CCRC. The methylation of the VHL, RASSF1, FHIT, and CDH1 genes was identified in 12, 56, 58.4, and 46.4% of primary tumors, respectively; that of at least one gene was in 84.1% of the samples. The hypermethylation of the RASSF1 gene was associated with late stages (p = 0.015 and the presence of metastases (p = 0.036; that of the CDH1 gene was related to the progression, invasion, and dissemination of primary tumors (p = 0.009, 0.039, and 0.002, respectively. The findings show it possible to use an analysis of abnormalities in the VHL gene and the methylation of the RASSF1 and CDH1 genes to develop a system of molecular genetic markers of RC.

  10. Aberrant methylation of the 3q25 tumor suppressor gene PTX3 in human esophageal squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    Jun-Xiong Wang; Yuan-Long He; Sheng-Tao Zhu; Shuo Yang; Shu-Tian Zhang

    2011-01-01

    AIM: To identify the novel methylation-silenced gene pentraxin 3 (PTX3) in esophageal squamous cell carcinoma (ESCC). METHODS: PTX3 mRNA expression was examined in six human ESCC cell lines, one human immortalized normal esophageal epithelial cell line, primary ESCC tumor tissue, and paired adjacent nontumor tissue using reverse transcription polymerase chain reaction (RT-PCR). Semi-quantitative immunohistochemistry was used to examine cellular localisation and protein levels. Methylation specific PCR and bisulphite genomic sequencing were employed to investigate the methylation of the candidate gene. RESULTS: In the majority of ESCC cell lines, we found that PTX3 expression was down-regulated due to gene promoter hypermethylation, which was further confirmed by bisulphite genomic sequencing. Demethyl-ation treatment with 5-aza-2'-deoxycytidine restored PTX3 mRNA expression in ESCC cell lines. Methylation was more common in tumor tissues (85%) than in adjacent nontumor tissues (25%) (P < 0 .01). CONCLUSION: PTX3 is down-regulated through promoter hypermethylation in ESCC, and could potentially serve as a biomarker of ESCC.

  11. Status of Intraductal Therapy for Ductal Carcinoma in Situ

    Science.gov (United States)

    Flanagan, Meghan; Love, Susan

    2010-01-01

    The intraductal approach is particularly appealing in the setting of ductal carcinoma in situ (DCIS), a preinvasive breast neoplasm that is thought to be entirely intraductal in its extent. Based on an emerging understanding of the anatomy of the ductal system as well as novel techniques to leverage the access accorded by the intraductal approach, researchers are actively exploring how ductal lavage, ductoscopy, and intraductal infusion of therapeutic agents may enhance breast cancer treatment. Both cytologic and molecular diagnostics continue to improve, and work is ongoing to identify the most effective diagnostic biomarkers for DCIS and cancer, although optimal targeting of the diseased duct remains an important consideration. Ductoscopy holds potential in detection of occult intraductal lesions, and ductoscopically guided lumpectomy could increase the likelihood of a more comprehensive surgical excision. Exciting pilot studies are in progress to determine the safety and feasibility of intraductal chemotherapy infusion. These studies are an important starting point for future investigations of intraductal ablative therapy for DCIS, because as our knowledge and techniques evolve, it is likely that DCIS may be the target most amenable to treatment by intraductal therapy. If such studies are successful, these approaches will allow an important and meaningful transformation in treatment options for women diagnosed with DCIS. PMID:21124756

  12. Current status of laparoscopic liver resection for hepatocellular carcinoma.

    Science.gov (United States)

    Guro, Hanisah; Cho, Jai Young; Han, Ho-Seong; Yoon, Yoo-Seok; Choi, YoungRok; Periyasamy, Mohan

    2016-06-01

    Laparoscopic liver resection (LLR) is becoming widely accepted for the treatment of hepatocellular carcinoma (HCC). Laparoscopic left lateral sectionectomy and minor laparoscopic liver resection are now considered standard approaches, especially for tumors located in the anterolateral segments of the liver. Laparoscopic left lateral sectionectomy in adult donors is also gaining acceptance for child liver transplantation in many centers. Major LLRs, including left hepatectomy and right hepatectomy, have been recently attempted. Laparoscopic donor hepatectomy is becoming more popular owing to increasing demand from young living donors who appreciate its minimal invasiveness and excellent cosmetic outcomes. Several centers have performed total laparoscopic donor right hepatectomy in adult-to-adult living donor liver transplantation. Many meta-analyses have shown that LLR is better than open liver resection in terms of short-term outcomes, principally cosmetic outcomes. Although no randomized control trials have compared LLR with open liver resection, the long-term oncologic outcomes were similar for both procedures in recent case-matched studies.

  13. Polymorphism of inflammatory genes and arsenic methylation capacity are associated with urothelial carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Wu, Chia-Chang [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Department of Urology, Taipei Medical University—Shuang Ho Hospital, Taipei, Taiwan (China); Huang, Yung-Kai [School of Oral Hygiene, College of Oral Medicine, Taipei Medical University, Taipei, Taiwan (China); Chung, Chi-Jung [Department of Health Risk Management, College of Public Health, China Medical University and Hospital, Taichung, Taiwan (China); Department of Medical Research, China Medical University Hospital, Taichung, Taiwan (China); Huang, Chao-Yuan; Pu, Yeong-Shiau [Department of Urology, National Taiwan University Hospital, College of Medicine National Taiwan University, Taipei, Taiwan (China); Shiue, Horng-Sheng [Department of Chinese Medicine, Chang Gung Memorial Hospital, Taipei, Taiwan (China); Lai, Li-An [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Lin, Ying-Chin [Department of Family Medicine, Shung Ho Hospital, Taipei Medical University, Taipei, Taiwan (China); Department of Health Examination, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan (China); Su, Chien-Tien [Department of Family Medicine, Taipei Medical University Hospital, Taipei, Taiwan (China); Hsueh, Yu-Mei, E-mail: ymhsueh@tmu.edu.tw [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Department of Public Health, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan (China)

    2013-10-01

    Chronic exposure to arsenic can generate reactive oxidative species, which can induce certain proinflammatory cytokines such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and interleukin-8 (IL-8). TNF-α, IL-6 and IL-8 have been shown to be involved in the development and progression of various cancers, including bladder cancer. This study aimed to investigate the joint effect of the polymorphism of TNF-α − 308 G/A, IL-6 − 174 G/C, IL-8 − 251 T/A and urinary arsenic profiles on urothelial carcinoma (UC) risk. This study evaluated 300 pathologically-confirmed cases of UC and 594 cancer-free controls. Urinary arsenic species were detected using high-performance liquid chromatography-linked hydride generator and atomic absorption spectrometry. The polymorphism of TNF-α − 308 G/A, IL-6 − 174 G/C and IL-8 − 251 T/A was determined using polymerase chain reaction-restriction fragment length polymorphism. The joint effects on UC risk were estimated by odds ratios and 95% confidence intervals using unconditional logistic regression. We found that the TNF-α − 308 A/A and IL-8 − 251 T/T polymorphisms were significantly associated with UC. Moreover, significant dose–response joint effect of TNF-α − 308 A/A or IL-8 − 251 T/T genotypes and arsenic methylation indices were seen to affect UC risk. The present results also showed a significant increase in UC risk in subjects with the IL-8 − 251 T/T genotype for each SD increase in urinary total arsenic and MMA%. In contrast, a significant decrease in UC risk was found in subjects who carried the IL-8 − 251 T/T genotype for each SD increase in DMA%. - Highlights: • Joint effect of the TNF-α -308 A/A genotype and urinary total arsenic affected UC. • Joint effect of the IL-8 -251 T/T genotype and urinary total arsenic affected UC. • Urinary total arsenic level, TNF-α -308 A/A and IL-8 -251 T/T genotype affected UC.

  14. Promoter methylation status of hMLH1,MGMT,and CDKN2A/p16 in colorectal adenomas

    Institute of Scientific and Technical Information of China (English)

    Vasiliki; Psofaki; Chryssoula; Kalogera; Nikolaos; Tzambouras; Dimitrios; Stephanou; Epameinondas; Tsianos; Konstantin; Seferiadis; Georgios; Kolios

    2010-01-01

    AIM:To investigate aberrant DNA methylation of CpG islands and subsequent low-or high-level DNA microsatellite instability(MSI)which is assumed to drive colon carcinogenesis. METHODS:DNA of healthy individuals,adenoma(tu-bular or villous/tubulovillous)patients,and colorectal carcinoma patients who underwent colonoscopy was used for assessing the prevalence of aberrant DNA methylation of human DNA mismatch repair gene mutator L homologue 1(hMLH1),Cyclin-dependent kinase inhibitor 2A(CDKN2A/p16),and O-6-methy...

  15. Chromosomal and methylation alterations in sporadic and familial adenomatous polyposis-related duodenal carcinomas.

    NARCIS (Netherlands)

    Berkhout, M.; Nagtegaal, I.D.; Cornelissen, S.J.B.; Dekkers, M.M.G.; Molengraft, F.J. van de; Peters, W.H.M.; Nagengast, F.M.; Krieken, J.H.J.M. van; Jeuken, J.W.M.

    2007-01-01

    Primary carcinomas of the small intestine are rare and the mechanism of their pathogenesis is poorly understood. Patients with familial adenomatous polyposis (FAP) have a high risk of developing duodenal carcinomas. The aim of this study is to gain more insight into the development of duodenal carci

  16. Relationship between methylation of MGMT gene promoter and cervical carcinoma%宫颈癌及宫颈上皮内瘤变MGMT基因甲基化及其相关性研究

    Institute of Scientific and Technical Information of China (English)

    陆欢; 陈莹蓉; 孙立奇; 陶志梅; 张甦; 邓再兴; 戴利成

    2014-01-01

    目的:探讨宫颈癌和宫颈上皮内瘤变(CIN)患者宫颈脱落细胞中O6-甲基鸟嘌呤- DNA甲基转移酶(MGMT)基因启动子区甲基化状态在宫颈癌临床早期诊断和筛查中的作用。方法选取对照组20例,CINI、CINII、CINIII期患者各50例和宫颈癌患者50例,采用甲基化特异性PCR检测宫颈脱落细胞中MGMT基因启动子区甲基化状态并进行比较。结果宫颈癌和CINI、II、III期患者MGMT基因启动子区甲基化阳性率分别为82.0%(41/50)、24.0%(12/50)、50.0%(25/50)和80.0%(40/50),而对照组未检测到MGMT基因启动子区甲基化。与对照组比,宫颈癌组和CINI、II、III期组MGMT基因启动子区甲基化阳性率的差异均有统计学意义(均P<0.05)。宫颈癌组中MGMT基因启动子甲基化与其临床分期及组织分化程度有显著相关性(均P<0.05)。结论 MGMT基因启动子区甲基化可能参与宫颈癌的发生、发展,有助于宫颈癌的早期辅助诊断和筛查。%Objective To investigate the methylation status of MGMT gene promoter in cervical carcinoma and cervical intraepithelial neoplasia (CIN). Methods Methylation status of MGMT gene promoter was detected with methylation specific PCR (MSP) in cervical exfoliated cel s from patients with CINI (n=50), CINII (n=50), CINIII (n=50), cervical carcinoma (n=50) and non- cancer controls(n=20). Results The promoter methylation rates of MGMT were 82.0%(41/50), 24.0%(12/50), 50.0%(25/50) and 80.0%(40/50) in cervical carcinoma, CIN I, CINII and CINIII, while no methylation was detected in control group (P<0.05). The methylation of MGMT gene was correlated with the clinicopathological characteristics and tissue differentiation of cervical carcinoma (P<0.05). Conclusion The methylation of MGMT gene promoter region is associated with cervical carcinoma, which may be used as potential marker for early diagnosis of cervical carcinoma.

  17. A CLINICAL STUDY ON CARCINOMA BREAST IN RELATION TO ER AND PR STATUS

    Directory of Open Access Journals (Sweden)

    Ramanaiah

    2015-09-01

    Full Text Available INTRODUCTION : Breast carcinoma is the most common malignant tumor and the leading cause of death in women worldwide 1 . It accounts for 15 % of all cancer deaths 2 . According to the World Health Organisation (WHO, approximately 70% of breast cancers occur in women with none of the known risk factors. Only about 5% of breast cancers are inherited. Various protocols are in use for the assessment of prognosis, and also to assist further management of these cases. Of various parameters, expression of hormonereceptors Estrogen receptor (ER and Progesterone receptor (PR ar e significant AIMS AND OBJECTIVES : To study the occurrence of ER and PR status in breast cancer patients attending S.V.R.R.G.G. Hospital. To correlate the expression of prognostic factors like age at presentation menarche, menopause, parity, tumor size, number of lymph nodes, metastasis histology, grading with ER and PR status. MATERIALS AND METHODS : This clinic opathological study of carcinoma breast was carried out in patients admitted to SVRRGG Hospital, Tirupati during the period from September 2011 to August 2013 after obtaining approval from scientific committee and ethical committee .Forty cases of breast carcinoma were taken into study. The clinical study done by interviewing, detailed examination and subjecting to relevant investigations and surgeries depending upon the stage of the disease. Excised specimen is sent for Histopathological examination in 10% formaline Reports of light microscopy (Hematoxilin and Eosin and immunohistochemistry on tumor histology including MBR (Modified Bloom Richardson grading and Estrogen and Progesterone status is analysed. CONCLUSION: In conclusion, ER and PR status correlates well with histopathological grading and other clinico - pathological parameters. Higher grade is associated with ER PR negativity. Hence. Immunohistochemical assessment of ER and PR status should be incorporated as a routine investigation. This along with

  18. Frequent methylation of the KLOTHO gene and overexpression of the FGFR4 receptor in invasive ductal carcinoma of the breast.

    Science.gov (United States)

    Dallol, Ashraf; Buhmeida, Abdelbaset; Merdad, Adnan; Al-Maghrabi, Jaudah; Gari, Mamdooh A; Abu-Elmagd, Muhammad M; Elaimi, Aisha; Assidi, Mourad; Chaudhary, Adeel G; Abuzenadah, Adel M; Nedjadi, Taoufik; Ermiah, Eramah; Alkhayyat, Shadi S; Al-Qahtani, Mohammed H

    2015-12-01

    Invasive ductal carcinoma of the breast is the most common cancer affecting women worldwide. The marked heterogeneity of breast cancer is matched only with the heterogeneity in its associated or causative factors. Breast cancer in Saudi Arabia is apparently an early onset with many of the affected females diagnosed before they reach the age of 50 years. One possible rationale underlying this observation is that consanguinity, which is widely spread in the Saudi community, is causing the accumulation of yet undetermined cancer susceptibility mutations. Another factor could be the accumulation of epigenetic aberrations caused by the shift toward a Western-like lifestyle in the past two decades. In order to shed some light into the molecular mechanisms underlying breast cancer in the Saudi community, we identified KLOTHO (KL) as a tumor-specific methylated gene using genome-wide methylation analysis of primary breast tumors utilizing the MBD-seq approach. KL methylation was frequent as it was detected in 55.3 % of breast cancer cases from Saudi Arabia (n = 179) using MethyLight assay. Furthermore, KL is downregulated in breast tumors with its expression induced following treatment with 5-azacytidine. The involvement of KL in breast cancer led us to investigate its relationship in the context of breast cancer, with one of the protagonists of its function, fibroblast growth factor receptor 4 (FGFR4). Overexpression of FGFR4 in breast cancer is frequent in our cohort and this overexpression is associated with poor overall survival. Interestingly, FGFR4 expression is higher in the absence of KL methylation and lower when KL is methylated and presumably silenced, which is suggestive of an intricate relationship between the two factors. In conclusion, our findings further implicate "metabolic" genes or pathways in breast cancer that are disrupted by epigenetic mechanisms and could provide new avenues for understanding this disease in a new context.

  19. Nuclear BMI-1 expression in laryngeal carcinoma correlates with lymph node pathological status

    Directory of Open Access Journals (Sweden)

    Allegra Eugenia

    2012-10-01

    Full Text Available Abstract Background The main cause of treatment failure and death in laryngeal squamous cell carcinoma is metastasis to the regional lymph nodes. The current clinical staging criteria fail to differentiate patients with occult metastasis from patients without metastasis. Identifying molecular markers of the disease might improve our understanding of the molecular mechanisms underlying the pathogenesis and development of laryngeal carcinoma and may help improve clinical staging and treatment. Methods Sixty-four previously untreated patients who underwent surgical excision of laryngeal squamous cell carcinoma with neck dissection were included in this study. The expression of B cell-specific Moloney murine leukemia virus integration site 1 (BMI-1 was examined immunohistochemically on formalin-fixed paraffin-embedded primary tissue specimens. Results Nuclear expression of BMI-1 (nBMI-1 was detected in 32 of the 64 tumors (50%, cytoplasmic expression of BMI-1 (cBMI-1 was detected in 22 (34.4%, and 10 tumors (15.6% showed no BMI-1 immunoreactivity. High nBMI-1 expression levels (≥10 were detected in 28 of the 32 (87.5% nBMI-1-positive patients. Multivariate analysis including age at diagnosis, grade, tumor location, TNM status, and nBMI-1 expression showed that a high nBMI-1 expression level was an independent prognostic factor for lymph node metastasis. Conclusion The expression of BMI-1 in patients with laryngeal carcinoma seems to correlate with lymph node metastasis.

  20. RESEARCH OF NUTRITIONAL AND IMMUNE STATUS IN PATIENTS WITH GALLBLADDER CARCINOMA RADICAL CHOLECYSTECTOMY

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective To inquire the nutritional and immune status in patients with gallbladder carcinoma before and after radical cholecystectomy.Methods The nutritional and immune status in patients with gallbladder carcinoma were assessed in 1 week before surgery, and on 3rd day, 7th day, 14th day and 21st day after operation, respectively.Results All of the nutritional parameters but the serum level of iron, TIBC and transferrin recovered within 3 week after operation. Remarkable decrease of serum IgG, IgA, IgM and C3, C4 complement, IL-2, CD4, CD4/CD8 ratio, and the remarkable increase of serum SIL-2R and CD8(P<0.01) on 3rd day after operation.Conclusion Adequate iron should be supplemented after the radical cholecystectomy for gallbladder carcinoma in the third postoperative week. Radical cholecystectomy with complete resection of the tumor and removal of lymph nodes played the important roles in the recovery of immune function.

  1. Stability of methylation markers in head and neck squamous cell carcinomas

    Science.gov (United States)

    Virani, S; Light, E; Peterson, LA; Sartor, MA; Taylor, JMG; McHugh, JB; Wolf, GT; Rozek, LS

    2016-01-01

    Background As cancer progresses, methylation patterns change to promote the tumorigenic phenotype. However, stability of methylation markers over time and the extent that biopsy samples are representative of larger tumor specimens are unknown. This information is critical for clinical use of such biomarkers. Methods Ninety-eight patients with tumor specimens from two time points were measured for DNA methylation in the promoter regions across four genes. Results There were no significant differences in overall methylation of CCNA1, DCC, CD1A or NDN within paired specimens (p-values= 0.56, 0.17, 0.66 and 0.58, respectively). All genes showed strong correlations between paired specimens across time. Methylation was most consistent for CCNA1 and NDN over time. Conclusions This report provides the first evidence that methylation markers measured in biopsy samples are representative of gene methylation in later specimens and suggests that biopsy markers could be representative biomarkers for use in defining personalized treatment utilizing epigenetic changes. PMID:26566081

  2. MGMT-Methylated Alleles Are Distributed Heterogeneously Within Glioma Samples Irrespective of IDH Status and Chromosome 10q Deletion.

    Science.gov (United States)

    Fontana, Laura; Tabano, Silvia; Bonaparte, Eleonora; Marfia, Giovanni; Pesenti, Chiara; Falcone, Rossella; Augello, Claudia; Carlessi, Nicole; Silipigni, Rosamaria; Guerneri, Silvana; Campanella, Rolando; Caroli, Manuela; Maria Sirchia, Silvia; Bosari, Silvano; Miozzo, Monica

    2016-06-26

    Several molecular markers drive diagnostic classification, prognostic stratification, and/or prediction of response to therapy in patients with gliomas. Among them, IDH gene mutations are valuable markers for defining subtypes and are strongly associated with epigenetic silencing of the methylguanine DNA methyltransferase (MGMT) gene. However, little is known about the percentage of MGMT-methylated alleles in IDH-mutated cells or the potential association between MGMT methylation and deletion of chromosome 10q, which encompasses the MGMT locus. Here, we quantitatively assessed MGMT methylation and IDH1 mutation in 208 primary glioma samples to explore possible differences associated with the IDH genotype. We also explored a potential association between MGMT methylation and loss of chromosome 10q. We observed that MGMT methylation was heterogeneously distributed within glioma samples irrespective of IDH status suggesting an incomplete overlap between IDH1-mutated and MGMT-methylated alleles and indicating a partial association between these two events. Moreover, loss of one MGMT allele did not affect the methylation level of the remaining allele. MGMT was methylated in about half of gliomas harboring a 10q deletion; in those cases, loss of heterozygosity might be considered a second hit leading to complete inactivation of MGMT and further contributing to tumor progression.

  3. Comparison of topical methyl aminolevulinate photodynamic therapy with cryotherapy or Fluorouracil for treatment of squamous cell carcinoma in situ: Results of a multicenter randomized trial.

    NARCIS (Netherlands)

    Morton, C.; Horn, M.; Leman, J.; Tack, B.; Bedane, C.; Tjioe, M.; Ibbotson, S.; Khemis, A.; Wolf, P.

    2006-01-01

    OBJECTIVE: To compare the efficacy, tolerability, and cosmetic outcome of photodynamic therapy (PDT) using topical methyl aminolevulinate with cryotherapy or topical fluorouracil for treatment of squamous cell carcinoma in situ. DESIGN: Randomized, placebo-controlled study, with follow-up at 3 and 1

  4. Methylation of a CpG Island within the Uroplakin Ib Promoter: A Possible Mechanism for Loss of Uroplakin lb Expression in Bladder Carcinoma

    Directory of Open Access Journals (Sweden)

    Andrea E. Varga

    2004-03-01

    Full Text Available Uroplakin Ib is a structural protein on the surface of urothelial cells. Expression of uroplakin Ib mRNA is reduced or absent in many transitional cell carcinomas (TCCs but molecular mechanisms underlying loss of expression remain to be determined. Analysis of the uroplakin Ib promoter identified a weak CpG island spanning the proximal promoter, exon 1, and the beginning of intron 1. This study examined the hypothesis that methylation of this CpG island regulates uroplakin Ib expression. Uroplakin Ib mRNA levels were determined by reverse transcription polymerase chain reaction and CpG methylation was assessed by bisulfite modification of DNA, PCR, and sequencing. A correlation was demonstrated in 15 TCC lines between uroplakin Ib mRNA expression and lack of CpG methylation. In support of a regulatory role for methylation, incubating uroplakin Ib-negative lines with 5-aza-2′ -deoxycytidine reactivated uroplakin Ib mRNA expression. A trend between uroplakin Ib mRNA expression and CpG methylation was also observed in normal urothelium and bladder carcinomas. In particular, loss of uroplakin Ib expression correlated with methylation of a putative Spi/NFκB binding motif. The data are consistent with the hypothesis that methylation of specific sites within the uroplakin Ib promoter may be an important factor in the loss of uroplakin Ib expression in TCCs.

  5. COX-2 gene expression in colon cancer tissue related to regulating factors and promoter methylation status

    Directory of Open Access Journals (Sweden)

    Lagerstedt Kristina

    2011-06-01

    Full Text Available Abstract Background Increased cyclooxygenase activity promotes progression of colorectal cancer, but the mechanisms behind COX-2 induction remain elusive. This study was therefore aimed to define external cell signaling and transcription factors relating to high COX-2 expression in colon cancer tissue. Method Tumor and normal colon tissue were collected at primary curative operation in 48 unselected patients. COX-2 expression in tumor and normal colon tissue was quantified including microarray analyses on tumor mRNA accounting for high and low tumor COX-2 expression. Cross hybridization was performed between tumor and normal colon tissue. Methylation status of up-stream COX-2 promoter region was evaluated. Results Tumors with high COX-2 expression displayed large differences in gene expression compared to normal colon. Numerous genes with altered expression appeared in tumors of high COX-2 expression compared to tumors of low COX-2. COX-2 expression in normal colon was increased in patients with tumors of high COX-2 compared to normal colon from patients with tumors of low COX-2. IL1β, IL6 and iNOS transcripts were up-regulated among external cell signaling factors; nine transcription factors (ATF3, C/EBP, c-Fos, Fos-B, JDP2, JunB, c-Maf, NF-κB, TCF4 showed increased expression and 5 (AP-2, CBP, Elk-1, p53, PEA3 were decreased in tumors with high COX-2. The promoter region of COX-2 gene did not show consistent methylation in tumor or normal colon tissue. Conclusions Transcription and external cell signaling factors are altered as covariates to COX-2 expression in colon cancer tissue, but DNA methylation of the COX-2 promoter region was not a significant factor behind COX-2 expression in tumor and normal colon tissue.

  6. Ancestral TCDD exposure promotes epigenetic transgenerational inheritance of imprinted gene Igf2: Methylation status and DNMTs.

    Science.gov (United States)

    Ma, Jing; Chen, Xi; Liu, Yanan; Xie, Qunhui; Sun, Yawen; Chen, Jingshan; Leng, Ling; Yan, Huan; Zhao, Bin; Tang, Naijun

    2015-12-01

    Ancestral TCDD exposure could induce epigenetic transgenerational phenotypes, which may be mediated in part by imprinted gene inheritance. The aim of our study was to evaluate the transgenerational effects of ancestral TCDD exposure on the imprinted gene insulin-like growth factor-2 (Igf2) in rat somatic tissue. TCDD was administered daily by oral gavage to groups of F0 pregnant SD rats at dose levels of 0 (control), 200 or 800 ng/kg bw during gestation day 8-14. Animal transgenerational model of ancestral exposure to TCDD was carefully built, avoiding sibling inbreeding. Hepatic Igf2 expression of the TCDD male progeny was decreased concomitantly with hepatic damage and increased activities of serum hepatic enzymes both in the F1 and F3 generation. Imprinted Control Region (ICR) of Igf2 manifested a hypermethylated pattern, whereas methylation status in the Differentially Methylated Region 2 (DMR2) showed a hypomethylated manner in the F1 generation. These epigenetic alterations in these two regions maintained similar trends in the F3 generation. Meanwhile, the expressions of DNA methyltransferases (DNMT1, DNMT3A and DNMT3B) changed in a non-monotonic manner both in the F1 and F3 generation. This study provides evidence that ancestral TCDD exposure may promote epigenetic transgenerational alterations of imprinted gene Igf2 in adult somatic tissue.

  7. CDH1基因甲基化与食管鳞癌的关系%Relationship between CDH1 gene methylation and esophageal squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    李永丽; 张立玮

    2012-01-01

    目的 探对食管鳞癌组织中CDH1基因启动子区甲基化的表达状况及其与烟酒史、肿瘤家族史的关系.方法 应用甲基化特异性聚合酶链反应(MSP)法检测食管鳞癌中CDH1基因启动子区甲基化情况.结果 食管鳞癌、癌旁及正常组织CDH1基因的甲基化阳性率分别为51.6%(32/62)、22.6%(14/62)和0(0/18).癌与癌旁组织比较,甲基化阳性率差异有统计学意义(P<0.01);癌和正常组织比较,甲基化阳性率差异有统计学意义(x2=15.484,P<0.01);癌旁与正常组织间差异无统计学意义(x2 =3.487,P>0.05).食管鳞癌CDH1基因甲基化情况与其性别、年龄、烟酒史和上消化道肿瘤家族史比较差异均无统计学意义(均P >0.05).结论 CDH1基因启动子区甲基化与食管鳞癌发生关系密切,在食管鳞癌中是一常见的表观遗传学事件.CDH1基因甲基化可能是一个独立的危险因素.%Objective To study CDH1 gene promoter methylation status in tissues of esophageal squamous cell' carcinomas(ESCC) and its relationship with tobacco and alcohol history, family history of cancer. Methods CDHl gene promoter methylation in ESCC Was detected by methylation-specific polymerase chain reaction (MSP) and was analyzed statistically. Results CDH1 gene methylation positive rates in tumor tissues, adjacent and normal tissues of ESCC were 51. 6%(32/62) ,22. 6%(14/62) and 0(0/18) .respectively,the difference between cancer and adjacent tissue was significant ( P 0. 05). Compared CDHl gene methylation in ESCC with sex,age,tobacco and alcohol history,family history of upper gastrointestinal tumors, there were no statistical significance (all P > 0. 05). Conclusion CDHl gene promoter methylation is closely related to the occurrence of ESCC, and it is a common epigenetic event in ESCC. CDHl gene methylation may be an independent risk factor.

  8. Evaluating the methylation status of CXCR4 promoter: A cost-effective and sensitive two-step PCR method.

    Science.gov (United States)

    Bianchessi, Valentina; Lauri, Andrea; Vigorelli, Vera; Toia, Martina; Vinci, Maria Cristina

    2017-02-15

    The chemokine receptor CXCR4 plays a key role in the bone marrow microenvironment maintenance and in the hematopoietic stem and progenitor cells migration. In addition, CXCR4 is expressed in a broad spectrum of solid tumors where its methylation state has been recently proposed as a biomarker for cancer prognosis. To evaluate methylation status of CXCR4 promoter we developed a sensitive, accurate, specific and cost-effective two-step PCR method that does not require any specific equipment other than a conventional real-time PCR instrument. The principle of the technique relies on a novel normalization strategy which allows the detection and quantification of small methylation differences among pre-amplified DNA samples deriving from low amount of starting material. In addition, the analysis of melting curve profiles of PCR products provides additional information about the methylation status of CpG sites in between the primers. Finally, the principle of this technique can potentially be adapted for the investigation of the methylation status of any other DNA region.

  9. Meningeal hemangiopericytomas: a clinicopathological study with emphasis on MGMT (O(6) -methylguanine-DNA methyltransferase) promoter methylation status.

    Science.gov (United States)

    Kakkar, Aanchal; Kumar, Anupam; Jha, Prerana; Goyal, Nishant; Mallick, Supriya; Sharma, Mehar Chand; Suri, Ashish; Singh, Manmohan; Kale, Shashank S; Julka, Pramod Kumar; Sarkar, Chitra; Suri, Vaishali

    2014-08-01

    Meningeal hemangiopericytomas (HPCs) are aggressive dural-based tumors, for which no prognostic or predictive marker has been identified. Gross total resection is treatment of choice, but not easily achieved; hence, alkylating agents like temozolomide (TMZ) are now being tried. O(6) -methylguanine-DNA methyltransferase (MGMT) promoter methylation has proven prognostic and predictive value in glioblastomas. This study evaluates MGMT promoter methylation in meningeal HPCs to determine its role in HPC oncogenesis and its association with patient outcome. Meningeal HPCs diagnosed between 2002 and 2011 were retrieved and clinicopathological features reviewed. MGMT promoter methylation status was assessed by methylation-specific polymerase chain reaction (MSP) and immunohistochemistry (IHC) for MGMT protein. HPCs accounted for 1.1% of all CNS tumors. Forty cases were analyzed; the majority were adults (mean age = 41.4 years). Seventy percent were primary and 30% were recurrent tumors; 60% were grade II and 40% were grade III. MGMT promoter methylation was identified in 45% of cases, including Grade II (54.2%) and Grade III (31.3%) (P = 0.203). Promoter methylation was significantly (P = 0.035) more frequent in primary (57.1%) than in recurrent (16.7%) tumors. No correlation was noted between MGMT promoter methylation by MSP and MGMT protein expression by IHC, or with progression-free survival. Thus, a significant proportion of HPCs demonstrate MGMT promoter methylation, suggesting possible susceptibility to TMZ. As promoter methylation is more frequent in primary tumors, TMZ may serve as a therapeutic option in residual primary tumors. Epigenetic inactivation of MGMT in HPCs necessitates the assessment of prognostic and predictive value of MGMT promoter methylation in HPCs in larger clinical trials.

  10. FOXM1 induces a global methylation signature that mimics the cancer epigenome in head and neck squamous cell carcinoma.

    Directory of Open Access Journals (Sweden)

    Muy-Teck Teh

    Full Text Available The oncogene FOXM1 has been implicated in all major types of human cancer. We recently showed that aberrant FOXM1 expression causes stem cell compartment expansion resulting in the initiation of hyperplasia. We have previously shown that FOXM1 regulates HELLS, a SNF2/helicase involved in DNA methylation, implicating FOXM1 in epigenetic regulation. Here, we have demonstrated using primary normal human oral keratinocytes (NOK that upregulation of FOXM1 suppressed the tumour suppressor gene p16(INK4A (CDKN2A through promoter hypermethylation. Knockdown of HELLS using siRNA re-activated the mRNA expression of p16(INK4A and concomitant downregulation of two DNA methyltransferases DNMT1 and DNMT3B. The dose-dependent upregulation of endogenous FOXM1 (isoform B expression during tumour progression across a panel of normal primary NOK strains (n = 8, dysplasias (n = 5 and head and neck squamous cell carcinoma (HNSCC cell lines (n = 11 correlated positively with endogenous expressions of HELLS, BMI1, DNMT1 and DNMT3B and negatively with p16(INK4A and involucrin. Bisulfite modification and methylation-specific promoter analysis using absolute quantitative PCR (MS-qPCR showed that upregulation of FOXM1 significantly induced p16(INK4A promoter hypermethylation (10-fold, P<0.05 in primary NOK cells. Using a non-bias genome-wide promoter methylation microarray profiling method, we revealed that aberrant FOXM1 expression in primary NOK induced a global hypomethylation pattern similar to that found in an HNSCC (SCC15 cell line. Following validation experiments using absolute qPCR, we have identified a set of differentially methylated genes, found to be inversely correlated with in vivo mRNA expression levels of clinical HNSCC tumour biopsy samples. This study provided the first evidence, using primary normal human cells and tumour tissues, that aberrant upregulation of FOXM1 orchestrated a DNA methylation signature that mimics the cancer methylome

  11. Molecular Profiling of Non-small Cell Lung Carcinomas : A Genome-wide DNA Methylation Analysis

    NARCIS (Netherlands)

    R. Hughes Carvalho (Rejane)

    2012-01-01

    textabstractDNA methylation is a signaling marker used by the cell to control gene expression, to keep genes silenced or active. It is an important part of what is called epigenetic controlling mechanisms (epi- Greek: επί- over, above, outer). We are just beginning to understand the intricate proces

  12. Correlation between aberrant methylation status of ras association domain family 1A and alteration of DNA methyltransferases, hepatitis B virus infection in hepatocellular carcinoma%原发性肝癌组织RASSF1A异常甲基化与DNMTs及HBV的关系

    Institute of Scientific and Technical Information of China (English)

    赵强子; 窦科峰; 祁志芳; 魏万礼; 杨雁灵; 李开宗

    2006-01-01

    目的探讨原发性肝癌ras相关结构域家族1A(ras association domain family 1A,RASSF1A)异常甲基化与肿瘤组织中DNA甲基转移酶(DNA methyltransferases,DNMTs)转录水平变化及HBV感染的关系.方法采用甲基化特异性PCR(methylation specific PCR,MSP)技术检测61例原发性肝癌组织RASSF1A基因异常甲基化情况,RT-PCR法检测其中三种主要DNA甲基转移酶(DNMT1、DNMT3A、DNMT3B)水平的变化.结果61例肝癌标本中RASSF1A基因启动子区异常甲基化45例(73.8%);HBV感染者47例(77.1%),其中MSP阳性32例;无HBV感染者14例,其中MSP阳性13例,RASSF1A基因异常甲基化与HBV感染之间的关系无统计学意义(x2=2.260,P=0.133).肝癌组、肝癌细胞系组及HBV感染组DNMTs表达水平与正常对照组相比均显著升高;组内比较显示肝癌组中DNMT3A、DNMT3B在MSP阳性患者中较阴性患者升高(分别为t=3.494,P<0.01;t=4.258,P<0.01),而DNMT1下降(t=3.221,P<0.05);HBV感染组中,MSP阳性组织其DNMT3B较阴性者升高(t=12.171,P<0.01).结论肝癌组织DNMTs水平变化与RASSF1A异常甲基化有密切关系;HBV感染与DNMT3B转录水平变化相关.

  13. Amyloid protein-mediated differential DNA methylation status regulates gene expression in Alzheimer's disease model cell line

    Energy Technology Data Exchange (ETDEWEB)

    Sung, Hye Youn; Choi, Eun Nam [Department of Biochemistry, School of Medicine, Ewha Womans University, 911-1 Mok-6-dong, Yangcheon-ku, Seoul 158-710 (Korea, Republic of); Ahn Jo, Sangmee [Department of Pharmacy, College of Pharmacy, Dankook University, San 29 Anseo-dong, Dongnam-gu, Cheonan-si, Chungnam 330-714 (Korea, Republic of); Oh, Seikwan [Department of Neuroscience and TIDRC, School of Medicine, Ewha Womans University, 911-1 Mok-6-dong, Yangcheon-ku, Seoul 158-710 (Korea, Republic of); Ahn, Jung-Hyuck, E-mail: ahnj@ewha.ac.kr [Department of Biochemistry, School of Medicine, Ewha Womans University, 911-1 Mok-6-dong, Yangcheon-ku, Seoul 158-710 (Korea, Republic of)

    2011-11-04

    Highlights: Black-Right-Pointing-Pointer Genome-wide DNA methylation pattern in Alzheimer's disease model cell line. Black-Right-Pointing-Pointer Integrated analysis of CpG methylation and mRNA expression profiles. Black-Right-Pointing-Pointer Identify three Swedish mutant target genes; CTIF, NXT2 and DDR2 gene. Black-Right-Pointing-Pointer The effect of Swedish mutation on alteration of DNA methylation and gene expression. -- Abstract: The Swedish mutation of amyloid precursor protein (APP-sw) has been reported to dramatically increase beta amyloid production through aberrant cleavage at the beta secretase site, causing early-onset Alzheimer's disease (AD). DNA methylation has been reported to be associated with AD pathogenesis, but the underlying molecular mechanism of APP-sw-mediated epigenetic alterations in AD pathogenesis remains largely unknown. We analyzed genome-wide interplay between promoter CpG DNA methylation and gene expression in an APP-sw-expressing AD model cell line. To identify genes whose expression was regulated by DNA methylation status, we performed integrated analysis of CpG methylation and mRNA expression profiles, and identified three target genes of the APP-sw mutant; hypomethylated CTIF (CBP80/CBP20-dependent translation initiation factor) and NXT2 (nuclear exporting factor 2), and hypermethylated DDR2 (discoidin domain receptor 2). Treatment with the demethylating agent 5-aza-2 Prime -deoxycytidine restored mRNA expression of these three genes, implying methylation-dependent transcriptional regulation. The profound alteration in the methylation status was detected at the -435, -295, and -271 CpG sites of CTIF, and at the -505 to -341 region in the promoter of DDR2. In the promoter region of NXT2, only one CpG site located at -432 was differentially unmethylated in APP-sw cells. Thus, we demonstrated the effect of the APP-sw mutation on alteration of DNA methylation and subsequent gene expression. This epigenetic regulatory

  14. HERV-K and LINE-1 DNA methylation and reexpression in urothelial carcinoma

    Directory of Open Access Journals (Sweden)

    Ulrike eKreimer

    2013-09-01

    Full Text Available Changes in DNA methylation frequently accompany cancer development. One prominent change is an apparently genome-wide decrease in methylcytosine that is often ascribed to DNA hypomethylation at retroelements comprising nearly half the genome. DNA hypomethylation may allow reactivation of retroelements, enabling retrotransposition and causing gene expression disturbances favoring tumor development. However, neither the extent of hypomethylation nor of retroelement reactivation are precisely known. We therefore assessed DNA methylation and expression of three major classes of retroelements (LINE-1, HERV-K and AluY in human urinary bladder cancer tissues and cell lines by pyrosequencing and quantitative reverse transcription–polymerase chain reaction, respectively. We found substantial global LINE-1 DNA hypomethylation in bladder cancer going along with a shift towards full-length LINE-1 expression. Thus, pronounced differences in LINE-1 expression were observed, which may be promoted, among others, by LINE-1 hypomethylation. Significant DNA hypomethylation was found at the HERV-K_22q11.23 proviral long terminal repeat (LTR in bladder cancer tissues but without reactivation of its expression. DNA methylation of HERVK17, essentially absent from normal urothelial cells, was elevated in cell lines from invasive bladder cancers. Accordingly, the faint expression of HERVK17 in normal urothelial cells disappeared in such cancer cell lines. Of 16 additional HERV-Ks, expression of 7 could be detected in the bladder, albeit generally at low levels. Unlike in prostate cancers, none of these showed significant expression changes in bladder cancer. In contrast, expression of the AluYb8 but not of the AluYa5 family was significantly increased in bladder cancer tissues. Collectively, our findings demonstrate a remarkable specificity of changes in expression and DNA methylation of retroelements in bladder cancer with a significantly different pattern from that

  15. Methylation status of the APC and RASSF1A promoter in cell-free circulating DNA and its prognostic role in patients with colorectal cancer.

    Science.gov (United States)

    Matthaios, Dimitrios; Balgkouranidou, Ioanna; Karayiannakis, Anastasios; Bolanaki, Helen; Xenidis, Nikolaos; Amarantidis, Kyriakos; Chelis, Leonidas; Romanidis, Konstantinos; Chatzaki, Aikaterini; Lianidou, Evi; Trypsianis, Grigorios; Kakolyris, Stylianos

    2016-07-01

    DNA methylation is the most frequent epigenetic alteration. Using methylation-specific polymerase chain reaction (MSP), the methylation status of the adenomatous polyposis coli (APC) and Ras association domain family 1 isoform A (RASSF1A) genes was examined in cell-free circulating DNA from 155 plasma samples obtained from patients with early and advanced colorectal cancer (CRC). APC and RASSF1A hypermethylation was frequently observed in both early and advanced disease, and was significantly associated with a poorer disease outcome. The methylation status of the APC and RASSF1A promoters was investigated in cell-free DNA of patients with CRC. Using MSP, the promoter methylation status of APC and RASSF1A was examined in 155 blood samples obtained from patients with CRC, 88 of whom had operable CRC (oCRC) and 67 had metastatic CRC (mCRC). The frequency of APC methylation in patients with oCRC was 33%. Methylated APC promoter was significantly associated with older age (P=0.012), higher stage (P=0.014) and methylated RASSF1A status (P=0.050). The frequency of APC methylation in patients with mCRC was 53.7%. In these patients, APC methylation was significantly associated with methylated RASSF1A status (P=0.016). The frequency of RASSF1A methylation in patients with oCRC was 25%. Methylated RASSF1A in oCRC was significantly associated with higher stage (P=0.021). The frequency of RASSF1A methylation in mCRC was 44.8%. Methylated RASSF1A in mCRC was associated with moderate differentiation (P=0.012), high levels of carcinoembryonic antigen (P=0.023) and methylated APC status (P=0.016). Patients with an unmethylated APC gene had better survival in both early (81±5 vs. 27±4 months, PAPC. Patients with an unmethylated RASSF1A gene had better survival in both early (71±6 vs. 46±8 months, PAPC and RASSF1A promoter methylation status and survival may be indicative of a prognostic role for these genes in CRC, which requires additional testing in larger studies.

  16. MGMT methylation analysis of glioblastoma on the Infinium methylation BeadChip identifies two distinct CpG regions associated with gene silencing and outcome, yielding a prediction model for comparisons across datasets, tumor grades, and CIMP-status

    NARCIS (Netherlands)

    P. Bady (Pierre); D. Sciuscio (Davide); A.C. Diserens; J. Bloch (Jocelyne); M.J. van den Bent (Martin); C. Marosi (Christine); P-Y. Dietrich (Pierre Yves); M. Weller (Michael); L. Mariani (Luigi); F.L. Heppner (Frank ); D.R. Mcdonald (David ); D. Lacombe (Denis); R. Stupp (Roger); M. Delorenzi (Mauro); M.E. Hegi (Monika)

    2012-01-01

    textabstractThe methylation status of the O6-methylguanine- DNA methyltransferase (MGMT) gene is an important predictive biomarker for benefit from alkylating agent therapy in glioblastoma. Recent studies in anaplastic glioma suggest a prognostic value for MGMT methylation. Investigation of pathogen

  17. DNA methylation by DNMT1 and DNMT3b methyltransferases is driven by the MUC1-C oncoprotein in human carcinoma cells.

    Science.gov (United States)

    Rajabi, H; Tagde, A; Alam, M; Bouillez, A; Pitroda, S; Suzuki, Y; Kufe, D

    2016-12-15

    Aberrant expression of the DNA methyltransferases (DNMTs) and disruption of DNA methylation patterns are associated with carcinogenesis and cancer cell survival. The oncogenic MUC1-C protein is aberrantly overexpressed in diverse carcinomas; however, there is no known link between MUC1-C and DNA methylation. Our results demonstrate that MUC1-C induces the expression of DNMT1 and DNMT3b, but not DNMT3a, in breast and other carcinoma cell types. We show that MUC1-C occupies the DNMT1 and DNMT3b promoters in complexes with NF-κB p65 and drives DNMT1 and DNMT3b transcription. In this way, MUC1-C controls global DNA methylation as determined by analysis of LINE-1 repeat elements. The results further demonstrate that targeting MUC1-C downregulates DNA methylation of the CDH1 tumor suppressor gene in association with induction of E-cadherin expression. These findings provide compelling evidence that MUC1-C is of functional importance to induction of DNMT1 and DNMT3b and, in turn, changes in DNA methylation patterns in cancer cells.

  18. Identification of human papillomavirus (HPV) 16 DNA integration and the ensuing patterns of methylation in HPV-associated head and neck squamous cell carcinoma cell lines.

    Science.gov (United States)

    Hatano, Takashi; Sano, Daisuke; Takahashi, Hideaki; Hyakusoku, Hiroshi; Isono, Yasuhiro; Shimada, Shoko; Sawakuma, Kae; Takada, Kentaro; Oikawa, Ritsuko; Watanabe, Yoshiyuki; Yamamoto, Hiroyuki; Itoh, Fumio; Myers, Jeffrey N; Oridate, Nobuhiko

    2017-04-01

    Recent studies showed that human papillomavirus (HPV) integration contributes to the genomic instability seen in HPV-associated head and neck squamous cell carcinoma (HPV-HNSCC). However, the epigenetic alterations induced after HPV integration remains unclear. To identify the molecular details of HPV16 DNA integration and the ensuing patterns of methylation in HNSCC, we performed next-generation sequencing using a target-enrichment method for the effective identification of HPV16 integration breakpoints as well as the characterization of genomic sequences adjacent to HPV16 integration breakpoints with three HPV16-related HNSCC cell lines. The DNA methylation levels of the integrated HPV16 genome and that of the adjacent human genome were also analyzed by bisulfite pyrosequencing. We found various integration loci, including novel integration sites. Integration loci were located predominantly in the intergenic region, with a significant enrichment of the microhomologous sequences between the human and HPV16 genomes at the integration breakpoints. Furthermore, various levels of methylation within both the human genome and the integrated HPV genome at the integration breakpoints in each integrant were observed. Allele-specific methylation analysis suggested that the HPV16 integrants remained hypomethylated when the flanking host genome was hypomethylated. After integration into highly methylated human genome regions, however, the HPV16 DNA became methylated. In conclusion, we found novel integration sites and methylation patterns in HPV-HNSCC using our unique method. These findings may provide insights into understanding of viral integration mechanism and virus-associated carcinogenesis of HPV-HNSCC.

  19. Effect of vitrification at the germinal vesicle stage on the global methylation status in mouse oocytes subsequently matured in vitro

    Institute of Scientific and Technical Information of China (English)

    Yan Jie; Zhang Lu; Wang Tianren; Li Rong; Liu Ping; Yan Liying; Qiao Jie

    2014-01-01

    Background It is still unclear whether the vitrification procedure itself is associated with the incidence of abnormal DNA methylation during oocytes vitrification.The purpose of this study was to evaluate the epigenetic profile of mouse oocytes,which went through vitrification either at a mature stage or at an immature stage following in vitro maturation (IVM) by analyzing the global DNA methylation.Methods Metaphase Ⅱ (M Ⅱ) stage and germinal vesicle (GV) stage oocytes were collected from adult female mice and were vitrified respectively.The M Ⅱ oocytes were assessed for cryo-survival and global DNA methylation.The GV oocytes were assessed for cryo-survival and only the surviving GV oocytes were cultured in vitro for subsequent assessment of global DNA methylation in mature oocytes.In vivo matured fresh M Ⅱ oocytes without undergoing vitrification were used as control.The level of global DNA methylation in the M Ⅱ oocytes was then examined by immunofluorescence using an anti-5-methylcytosine (anti-5-MeC) monoclonal antibody and fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG under a laser scanning confocal microscope.Results In terms of the effect of vitrification on global DNA methylation status in matured oocytes,in the M Ⅱ-v group,all the examined oocytes (90/90) were found with hypermethylation,including 63.3% (57/90) of them displaying DNA methylation of a very high level,25.6% (23/90) with a high level,and 11.1% (10/90) with an intermediate level,whereas in the GV-v group,all the matured oocytes (129/129) were also examined with hypermethylation,including 67.4% (87/129) of them displaying DNA methylation of a very high level,23.3% (30/129) with a high level,and 9.3% (12/129) with an intermediate level.Statistically,it was similar between both groups,which were similar to the control:68.6% (83/121) of fresh M Ⅱ oocytes displayed DNA methylation of a very high level,21.5% (26/121) with a high level,and 9.9%(12

  20. Association between the methylation status of the MGMT promoter in bone marrow specimens and chemotherapy outcomes of patients with acute myeloid leukemia.

    Science.gov (United States)

    Hong, Qingxiao; Chen, Xiaoying; Ye, Huadan; Zhou, Annan; Gao, Yuting; Jiang, Danjie; Wu, Xiaodong; Tian, Bingru; Chen, Youfen; Wang, Ming; Xie, Jiping; Xia, Yongming; Duan, Shiwei

    2016-04-01

    The O(6)-methylguanine-DNA methyltransferase (MGMT) gene is a tumor suppressor gene that is associated with the risk of developing acute myeloid leukemia (AML). However, the association between the methylation status of the MGMT promoter and the chemotherapeutic outcomes of patients with AML remains unknown. In the present study, 30 bone marrow samples derived from patients with AML were collected prior and subsequent to chemotherapy. The methylation status of the MGMT promoter in the bone marrow specimens was determined by methylation-specific polymerase chain reaction. The results indicated that the methylation status of the MGMT promoter was influenced by different chemotherapeutic regimens. The MGMT methylation status of M4 patients (3 out of 6) were more chemosensitive, compared with that of patients with other AML subtypes (M1, 1 out of 3; M2, 0 out of 8; M3, 3 out of 7; M5, 0 out of 3; and M6, 1 out of 3). Age-based analysis revealed that the group aged ≤60 years (7 out of 24 patients) exhibited more methylation changes than patients aged >60 years (1 out of 6). Male patients (4 out of 13) were more susceptible to chemotherapy-induced methylation changes than female patients (4 out of 17). Thus, the methylation status of the MGMT promoter may serve as a potential biomarker to predict the therapeutic outcomes in male AML patients. However, further studies in larger sample sets are required to confirm the present findings.

  1. KRAS, BRAF and PIK3CA status in squamous cell anal carcinoma (SCAC.

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    Andrea Casadei Gardini

    Full Text Available Anti-EGFR therapy appears to be a potential treatment option for squamous cell anal carcinoma (SCAC. KRAS mutation is a rare event in SCAC, indicating the absence of the principal mechanism of resistance to this type of therapy. However, no information is available from the literature regarding the status of BRAF or PIK3CA in this cancer type. We analysed KRAS, BRAF and PIK3CA status in SCAC patients in relation to the clinical-pathological characteristics of patients and to the presence of the human papilloma virus (HPV. One hundred and three patients were treated with the Nigro scheme for anal cancer from March 2001 to August 2012. Fifty patients were considered for the study as there was insufficient paraffin-embedded tumour tissue to perform molecular analysis the remaining 53. DNA was extracted from paraffin-embedded sections. KRAS, BRAF and PIK3CA gene status and HPV genotype were evaluated by pyrosequencing. KRAS and BRAF genes were wild-type in all cases. Conversely, PIK3CA gene was found to be mutated in 11 (22% cases. In particular, 8 mutations occurred in exon 9 and 3 in exon 20 of the PIK3CA gene. These findings suggest that SCAC could potentially respond to an anti-EGFR drug. PIK3CA mutation may be involved in the process of carcinogenesis in some cases of SCAC.

  2. CLINICAL STUDY OF CORRELATION BETWEEN ESTROGEN RECEPTOR (ER AND PROGESTERONE RECEPTOR (PR STATUS AND PATIENT CHARACTERISTICS IN CARCINOMA BREAST

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    Kiran Kumar

    2016-03-01

    Full Text Available BACKGROUND AND INTRODUCTION Breast carcinoma is the first among women of all races and Hispanic origin populations; most common carcinoma in women and account for 25% of all female cancers, which is more than twice the prevalence of cancer in women at any other site. Recent information suggests globally breast carcinoma is the second leading cause of cancer related death and is the most common cancer among women excluding the non-melanoma skin cancer. Estrogen and Progesterone play a central role in regulating growth kinetics of a variety of epithelial lining like in breast and endometrium and are powerful predictive markers in carcinoma of breast. AIMS AND OBJECTIVES To study the correlation between ER and PR status and patient characteristics like age of patient, menarche, menopause, parity, tumour size, lymph node status and tumour grade in carcinoma breast patient. MATERIALS AND METHODS Forty patients were included in this study who were admitted with breast carcinoma, the patient characteristics are correlated with ER and PR status studied by immunohistochemistry. This study was done over the period from October 2013 to September 2015. RESULTS Patients with both ER/PR positive are better differentiated, low grade, small sized tumours and with less chance of lymph node metastasis. Progesterone receptor negative status is associated with lymph node metastasis independent of other patient characteristics. Grade 3 and 4 tumours showed lower level of Estrogen and Progesterone receptors irrespective of age. CONCLUSION ER/PR positive group were better differentiated, showed better prognosis with hormonal therapy. They were less aggressive, low lymph node metastasis when compared with patients with either of the receptors negative or both the receptors negative. Hence immunohistochemistry is a must as it provides valuable prognostic, predictive and therapeutic information.

  3. The predictive but not prognostic value of MGMT promoter methylation status in elderly glioblastoma patients: a meta-analysis.

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    An-an Yin

    Full Text Available BACKGROUND: The clinical implication of O6-methylguanine-DNA methyltransferase (MGMT promoter status is ill-defined in elderly glioblastoma patients. Here we report a meta-analysis to seek valid evidence for its clinical relevance in this subpopulation. METHODS: Literature were searched and reviewed in a systematic manner using the PubMed, EMBASE and Cochrane databases. Studies investigating the association between MGMT promoter status and survival data of elderly patients (≥65 years were eligible for inclusion. RESULTS: Totally 16 studies were identified, with 13 studies included in the final analyses. The aggregate proportion of MGMT promoter methylation in elderly patients was 47% (95% confidence interval [CI]: 42-52%, which was similar to the value for younger patients. The analyses showed differential effects of MGMT status on overall survival (OS of elderly patients according to assigned treatments: methylated vs. unmethylated: (1 temozolomide (TMZ-containing therapies: hazard ratio [HR] 0.49, 95% CI 0.41-0.58; (2 TMZ-free therapies: HR 0.97, 95% CI 0.77-1.21. More importantly, a useful predictive value was observed by an interaction analysis: TMZ-containing therapies vs. RT alone: (1 methylated tumors: HR 0.48, 95% CI 0.36-0.65; (2 unmethylated tumors: HR 1.14; 95% CI 0.90-1.44. CONCLUSION: The meta-analysis reports an age-independent presence of MGMT promoter methylation. More importantly, the study encouraged routine testing of MGMT promoter status especially in elderly glioblastoma patients by indicating a direct linkage between biomarker test and individual treatment decision. Future studies are needed to justify the mandatory testing in younger patients.

  4. Methylation status of imprinting centers for H19/IGF2 and SNURF/SNRPN in primate embryonic stem cells.

    Science.gov (United States)

    Mitalipov, Shoukhrat; Clepper, Lisa; Sritanaudomchai, Hathaitip; Fujimoto, Akihisa; Wolf, Don

    2007-03-01

    Embryonic stem cells (ESCs) hold promise for cell and tissue replacement approaches to treating human diseases based on their capacity to differentiate into a wide variety of somatic cells and tissues. However, long-term in vitro culture and manipulations of ESCs may adversely affect their epigenetic integrity, including imprinting. We have recently reported aberrant biallelic expression of IGF2 and H19 in several rhesus monkey ESC lines, whereas SNRPN and NDN were normally imprinted and expressed predominantly from the paternal allele. The dysregulation of IGF2 and H19 that is associated with tumorigenesis in humans may result from improper maintenance of allele-specific methylation patterns at an imprinting center (IC) upstream of H19. To test this possibility, we performed methylation analysis of several monkey ESC lines by genomic bisulfite sequencing. We investigated methylation profiles of CpG islands within the IGF2/H19 IC harboring the CTCF-6 binding site. In addition, the methylation status of the IC within the promoter/exon 1 of SNURF/SNRPN known as the Prader-Willi syndrome IC was examined. Our results demonstrate abnormal hypermethylation within the IGF2/H19 IC in all analyzed ESC lines, whereas the SNURF/SNRPN IC was differentially methylated, consistent with monoallelic expression.

  5. DNA methylation-dependent regulation of TrkA, TrkB, and TrkC genes in human hepatocellular carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Jin, Wook [Laboratory of Molecular Disease and Cell Regulation, Lee Gil Ya Cancer and Diabetes Institute, Gachon University of Medicine and Science, Incheon (Korea, Republic of); Lee, Jong-Joo [Department of Environmental Medical Biology, Institute of Tropical Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, 250 Seongsanno, Seodaemun-gu, Seoul 120-752 (Korea, Republic of); Kim, Min Soo [Laboratory of Molecular Disease and Cell Regulation, Lee Gil Ya Cancer and Diabetes Institute, Gachon University of Medicine and Science, Incheon (Korea, Republic of); Son, Byung Ho [Department of Surgery, Kangbuk Samsung Hospital, Sungkyunkwan University, School of Medicine, 108 Pyung-dong, Jongro-gu, Seoul 110-746 (Korea, Republic of); Cho, Yong Kyun, E-mail: choyk2004@hanmail.net [Division of Gastroenterology, Department of Internal Medicine, Kangbuk Samsung Hospital, Sungkyunkwan University, School of Medicine, 108 Pyung-dong, Jongro-gu, Seoul 110-746 (Korea, Republic of); Kim, Hyoung-Pyo, E-mail: kimhp@yuhs.ac [Department of Environmental Medical Biology, Institute of Tropical Medicine, Brain Korea 21 Project for Medical Science, Yonsei University College of Medicine, 250 Seongsanno, Seodaemun-gu, Seoul 120-752 (Korea, Republic of)

    2011-03-04

    Research highlights: {yields} Expression of TrkA, TrkB, and TrkC is significantly elevated in human hepatocellular carcinoma. {yields} Downregulation of Trks is correlated with their promoter hypermethylation. {yields} Inhibiting DNA methylation restored expression of Trks in normal liver cell lines. {yields} Trks promote the proliferation of hepatocellular carcinoma. {yields} Trks induce expression of the metastatic regulator, Twist. -- Abstract: The tropomyosin-related kinase (Trk) family of neurotrophin receptors, TrkA, TrkB and TrkC, has been implicated in the growth and survival of human cancers. Here we report that Trks are frequently overexpressed in hepatocellular carcinoma (HCC) from patients and human liver cancer cell lines. To unravel the underlying molecular mechanism(s) for this phenomenon, DNA methylation patterns of CpG islands in TrkA, TrkB, and TrkC genes were examined in normal and cancer cell lines derived from liver. A good correlation was observed between promoter hypermethylation and lower expression of TrkA, TrkB, and TrkC genes, which was supported by the data that inhibiting DNA methylation with 5-azacytidine restored expression of those genes in normal liver cell lines. Furthermore, Trks promoted the proliferation of HepG2 and induced expression of the metastatic regulator, Twist. These results suggest that Trks may contribute to growth and metastasis of liver cancer.

  6. Association of Arsenic Methylation Capacity with Developmental Delays and Health Status in Children: A Prospective Case–Control Trial

    Science.gov (United States)

    Hsueh, Yu-Mei; Chen, Wei-Jen; Lee, Chih-Ying; Chien, Ssu-Ning; Shiue, Horng-Sheng; Huang, Shiau-Rung; Lin, Ming-I.; Mu, Shu-Chi; Hsieh, Ru-Lan

    2016-11-01

    This case–control study identified the association between the arsenic methylation capacity and developmental delays and explored the association of this capacity with the health status of children. We recruited 120 children with developmental delays and 120 age- and sex-matched children without developmental delays. The health status of the children was assessed using the Pediatric Quality of Life Inventory (PedsQL) and Pediatric Outcomes Data Collection Instrument (PODCI). The arsenic methylation capacity was determined by the percentages of inorganic arsenic (InAs%), monomethylarsonic acid (MMAV%), and dimethylarsinic acid (DMAV%) through liquid chromatography and hydride generation atomic absorption spectrometry. Developmental delays were significantly positively associated with the total urinary arsenic concentration, InAs%, and MMAV%, and was significantly negatively associated with DMAV% in a dose-dependent manner. MMAV% was negatively associated with the health-related quality of life (HRQOL; ‑1.19 to ‑1.46, P < 0.01) and functional performance (‑0.82 to ‑1.14, P < 0.01), whereas DMAV% was positively associated with HRQOL (0.33–0.35, P < 0.05) and functional performance (0.21–0.39, P < 0.01–0.05) in all children and in those with developmental delays. The arsenic methylation capacity is dose-dependently associated with developmental delays and with the health status of children, particularly those with developmental delays.

  7. Methylation of Tumor Suppressor Genes and Clinical Significance in Esophageal Squamous Cell Carcinoma%多种抑癌基因在食管鳞状细胞癌中的甲基化状态及其临床意义

    Institute of Scientific and Technical Information of China (English)

    王长春; 毛伟敏; 凌志强

    2012-01-01

    背景:抑癌基因甲基化与食管癌相关,目前多种抑癌基因与肿瘤家族史相关性的报道少见.目的:研究多种抑癌基因在食管鳞状细胞癌(ESCC)中的甲基化状态及其临床意义.方法:选取2010年2~7月浙江省肿瘤医院76例ESCC患者.应用MSP技术检测肿瘤组织和相应癌旁正常组织中APC、RARβ2、CDH1、p16INK4a、RASSF1A等5个抑癌基因的甲基化状态,并分析抑癌基因甲基化状态与肿瘤家族史的关系及其对预后的影响.结果:ESCC组织APC、RARβ2、CDH1、p16INK4a、RASSF1A的甲基化率均显著高于相应癌旁正常组织(P<0.05).ESCC组织中APC、RAR32、CDH1、RASSF1A甲基化与肿瘤家族史相关(P<0.05);CDH1、RASSF1A甲基化患者的生存期明显低于非甲基化患者(P=0.015、P=0.016).结论:ESCC患者存在抑癌基因APC、RARβ2、CDH1、p16INK4a、RASSF1A高甲基化;且APC、RARβ2、CDH1、RASSF1A甲基化与肿瘤家族史显著相关,CDH1、RASSF1A甲基化患者的预后可能较差.%Background: There is a correlation between methylation of tumor suppressor genes and esophageal carcinoma, but studies on relationship between methylation of tumor suppressor genes and family history of carcinoma are few. Aims: To study the methylation of tumor suppressor genes and clinical significance in esophageal squamous cell carcinoma (ESCC). Methods: Seventy-six patients with ESCC from Feb. To July 2010 at Zhejiang Cancer Hospital were enrolled. MSP technique was used to detect methylation status of APC, RAR(32, CDH1, plG?4" and RASSF1A in tumor tissue and paracancerous normal tissue. Relationship of methylation status of tumor suppressor genes with family history of carcinoma was analyzed, and their influences on prognosis was estimated. Results: The methylation rates of APC, RAR|32, CDH1, pjfjiM?a and RASSF1A were significantly higher in tumor tissue than those in paracancerous normal tissue (P<0.05). Methylation of APC, RARp2, CDH1, RASSF1A genes in

  8. Methylation profile of the promoter CpG islands of 14"drug-resistance" genes in hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Sheng Ding; Bang-Dong Gong; Jian Yu; Jun Gu; Hong-Yu Zhang; Zu-Bin Shang; Qi Fei; Peng Wang; Jing-De Zhu

    2004-01-01

    AIM: To establish the DNA methylation patterns of the promoter CpG islands of 14 "drug-resistance" genes in hepatocellular carcinoma (HCC).METHODS: The methylation specific polymerase chain reaction in conjunction with sequencing verification was used to establish the methylation patterns of the 14 genes in the liver tissues of four healthy liver donors, as well as tumor and the paired non-cancerous tissues of 30 HCC patients.RESULTS: While 11 genes (ATP-binding cassette, sub-family G (WHITE), member 2(ABCG2), activating transcription factor (ATF2), beta-2-microglobulin (B2M), deoxycytidine kinase (DCK), occludin (OCLN), v-raf-1 murine leukemia viral oncogene homolog (RAF1), ralA binding protein 1 (RALBP1),splicing factor (45 kD) (SPF45), S-phase kinase-associated protein 2 (p45) (SKP2), tumor protein p53 (Li-Fraumeni syndrome) (TP53) and topoisomerase (DNA) Ⅱ beta (TOP2B))maintained the unmethylated patterns, three genes displayed to various extents the hypermethylation state in tumor tissues in comparison with the normal counterparts. The catalase (CAT) was hypermethylated in tumor and the neighboring non-cancerous tissue of one case (3.3%). Both glutathione S-transferase pi (GSTpi) (80%, 24/30 in tumor and 56.7%,17/30 in the paired non-cancerous tissues) and cystic fibrosis transmembrane conductance regulator, ATP-binding cassette (sub-family C, member 7) (CFTR) (77%, 23/30 in tumor and 50%, 15/30 in the paired non-cancerous tissues) genes were prevalently hypermethylated in HCC as well as their neighboring non-cancerous tissues. No significant difference in the hypermethylation occurrence was observed between the HCC and its neighboring non-cancerous tissues.CONCLUSION: Hypermethylation of promoter CpG islands of both CFTR and GSTpi genes occurs prevalently in HCC,which may correlate with the low expression of these two genes at the mRNA level and has the profound etiological and clinical implications. It is likely to be specific to the early phase of HCC

  9. Molecular Features and Methylation Status in Early Onset (≤40 Years Colorectal Cancer: A Population Based, Case-Control Study

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    Giulia Magnani

    2015-01-01

    Full Text Available Colorectal cancer is usually considered a disease of the elderly. However, a small fraction of patients develops colorectal cancer earlier. The aim of our study was to define the frequency of known hereditary colorectal syndromes and to characterise genetic and epigenetic features of early nonhereditary tumors. Thirty-three patients ≤40 years with diagnosis of colorectal cancer and 41 patients with disease at >60 years of age were investigated for MSI, Mismatch Repair proteins expression, KRAS and BRAF mutations, hypermethylation, and LINE-1 hypomethylation. Detection of germline mutations was performed in Mismatch Repair, APC and MUTYH genes. Early onset colorectal cancer showed a high incidence of hereditary forms (18%. KRAS mutations were detected in 36% of early nonhereditary tumors. Early onset colorectal cancer disclosed an average number of methylated genes significantly lower when compared to the controls (p=0.02. Finally both of the two groups were highly methylated in ESR1, GATA5, and WT1 genes and were similar for LINE-1 hypomethylation. The genetic make-up of carcinomas differs from young to elderly patients. Early onset tumors showed more frequently a constitutional defective of Mismatch Repair System and a minor number of methylated genes. Hypermethylation of ESR1, GATA5, and WT1 genes suggests possible markers in the earlier diagnosis of colorectal tumorigenesis.

  10. Molecular Features and Methylation Status in Early Onset (≤40 Years) Colorectal Cancer: A Population Based, Case-Control Study

    Science.gov (United States)

    Magnani, Giulia; Furlan, Daniela; Sahnane, Nora; Reggiani Bonetti, Luca; Domati, Federica; Pedroni, Monica

    2015-01-01

    Colorectal cancer is usually considered a disease of the elderly. However, a small fraction of patients develops colorectal cancer earlier. The aim of our study was to define the frequency of known hereditary colorectal syndromes and to characterise genetic and epigenetic features of early nonhereditary tumors. Thirty-three patients ≤40 years with diagnosis of colorectal cancer and 41 patients with disease at >60 years of age were investigated for MSI, Mismatch Repair proteins expression, KRAS and BRAF mutations, hypermethylation, and LINE-1 hypomethylation. Detection of germline mutations was performed in Mismatch Repair, APC and MUTYH genes. Early onset colorectal cancer showed a high incidence of hereditary forms (18%). KRAS mutations were detected in 36% of early nonhereditary tumors. Early onset colorectal cancer disclosed an average number of methylated genes significantly lower when compared to the controls (p = 0.02). Finally both of the two groups were highly methylated in ESR1, GATA5, and WT1 genes and were similar for LINE-1 hypomethylation. The genetic make-up of carcinomas differs from young to elderly patients. Early onset tumors showed more frequently a constitutional defective of Mismatch Repair System and a minor number of methylated genes. Hypermethylation of ESR1, GATA5, and WT1 genes suggests possible markers in the earlier diagnosis of colorectal tumorigenesis. PMID:26557847

  11. Prognostic implications of FGFR1 and MYC status in esophageal squamous cell carcinoma

    Science.gov (United States)

    Kwon, Dohee; Yun, Ji Yun; Keam, Bhumsuk; Kim, Young Tae; Jeon, Yoon Kyung

    2016-01-01

    AIM To investigate the clinicopathological features and prognostic implications of combined MYC and fibroblast growth factor receptor 1 (FGFR1) status in esophageal squamous cell carcinomas (ESCCs). METHODS All patients with ESCC (n = 180) underwent surgical resection at Seoul National University Hospital sometime between 2000 and 2013. A tissue microarray was constructed using cores obtained from representative tumor areas of formalin-fixed, paraffin-embedded tissue blocks. FGFR1 and MYC copy numbers were quantified using fluorescence in situ hybridization. The level of MYC expression was determined using immunohistochemistry. FGFR1 and MYC amplification status was compared between primary and metastatic lymph nodes. Univariate and multivariate survival analyses were performed according to adjuvant therapy status. RESULTS FGFR1 and MYC amplifications were observed in 21.4% (37/173) and 54.2% (91/168) of patients, respectively, while MYC expression was observed in 58.9% (106/180) of patients. There was a positive correlation between MYC amplification and overexpression (P = 0.002). Although FGFR1 amplification was not associated with MYC amplification or expression, 12.3% (20/163) of patients exhibited both FGFR1 amplification and MYC expression. There was also a correlation in FGFR1 amplification status between matched primary tumors and metastatic lymph nodes (P FGFR1 amplification was an independent predictor for prolonged OS in all patients (P = 0.029) and in those who did not receive adjuvant therapy (P = 0.013). Combined FGFR1 amplification and MYC expression predicted better OS in patients who did not receive adjuvant therapy (P = 0.034) but not in those who did receive adjuvant therapy. CONCLUSION FGFR1 amplification and MYC expression have prognostic implications in resected ESCCs with respect to adjuvant therapy. The role of FGFR1-targeted therapy in ESCC remains to be explored. PMID:27956804

  12. Correlation of primary tumor size and axillary nodal status with tumor suppressor gene p53 in breast carcinoma

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    Topić Brano

    2002-01-01

    Full Text Available Correlation of standard path morphological prognostic parameters, primary tumor size and axillary nodal status with new prognostic factor in breast carcinoma: tumor suppressor gene p53 was analyzed. The studied sample included 65 women who underwent surgery for breast carcinoma at the Surgical Clinic of Clinical Center Banja Luka, from January 1st 1997 till January 1st 1999. Statistical data analysis was performed and correlation of prognostic factors was determined. The majority of authors in this field agree that the primary tumor size and axillary nodal status are the two most important prognostic factors. These factors are the best predictors of prognosis and survival of women who had the tumor and were operated on. Tumor markers were immunohistochemically determined in the last ten years and, according to the majority of authors, are still considered the additional or relative prognostic factors in breast carcinoma. Their prognostic value and significance increase almost daily. Most frequently determined tumor markers are bcl-2, pS2, Ki-67 and p53. There was a positive, directly proportional relationship between primary tumor size and tumor suppressor gene p53, but there was no positive correlation between the axillary nodal status and tumor suppressor gene p53. Significance of determination of new tumor markers as the prognostic factors was emphasized. These markers represent a powerful tool in the early detection and prevention of breast carcinoma.

  13. A combination of TERT promoter mutation and MGMT methylation status predicts clinically relevant subgroups of newly diagnosed glioblastomas.

    Science.gov (United States)

    Arita, Hideyuki; Yamasaki, Kai; Matsushita, Yuko; Nakamura, Taishi; Shimokawa, Asanao; Takami, Hirokazu; Tanaka, Shota; Mukasa, Akitake; Shirahata, Mitsuaki; Shimizu, Saki; Suzuki, Kaori; Saito, Kuniaki; Kobayashi, Keiichi; Higuchi, Fumi; Uzuka, Takeo; Otani, Ryohei; Tamura, Kaoru; Sumita, Kazutaka; Ohno, Makoto; Miyakita, Yasuji; Kagawa, Naoki; Hashimoto, Naoya; Hatae, Ryusuke; Yoshimoto, Koji; Shinojima, Naoki; Nakamura, Hideo; Kanemura, Yonehiro; Okita, Yoshiko; Kinoshita, Manabu; Ishibashi, Kenichi; Shofuda, Tomoko; Kodama, Yoshinori; Mori, Kanji; Tomogane, Yusuke; Fukai, Junya; Fujita, Koji; Terakawa, Yuzo; Tsuyuguchi, Naohiro; Moriuchi, Shusuke; Nonaka, Masahiro; Suzuki, Hiroyoshi; Shibuya, Makoto; Maehara, Taketoshi; Saito, Nobuhito; Nagane, Motoo; Kawahara, Nobutaka; Ueki, Keisuke; Yoshimine, Toshiki; Miyaoka, Etsuo; Nishikawa, Ryo; Komori, Takashi; Narita, Yoshitaka; Ichimura, Koichi

    2016-08-08

    The prognostic impact of TERT mutations has been controversial in IDH-wild tumors, particularly in glioblastomas (GBM). The controversy may be attributable to presence of potential confounding factors such as MGMT methylation status or patients' treatment. This study aimed to evaluate the impact of TERT status on patient outcome in association with various factors in a large series of adult diffuse gliomas. We analyzed a total of 951 adult diffuse gliomas from two cohorts (Cohort 1, n = 758; Cohort 2, n = 193) for IDH1/2, 1p/19q, and TERT promoter status. The combined IDH/TERT classification divided Cohort 1 into four molecular groups with distinct outcomes. The overall survival (OS) was the shortest in IDH wild-type/TERT mutated groups, which mostly consisted of GBMs (P MGMT methylation on survival of patients with GBM, samples from a combined cohort of 453 IDH-wild-type GBM cases treated with radiation and temozolomide were analyzed. A multivariate Cox regression model revealed that the interaction between TERT and MGMT was significant for OS (P = 0.0064). Compared with TERT mutant-MGMT unmethylated GBMs, the hazard ratio (HR) for OS incorporating the interaction was the lowest in the TERT mutant-MGMT methylated GBM (HR, 0.266), followed by the TERT wild-type-MGMT methylated (HR, 0.317) and the TERT wild-type-MGMT unmethylated GBMs (HR, 0.542). Thus, patients with TERT mutant-MGMT unmethylated GBM have the poorest prognosis. Our findings suggest that a combination of IDH, TERT, and MGMT refines the classification of grade II-IV diffuse gliomas.

  14. Using the apparent diffusion coefficient to identifying MGMT promoter methylation status early in glioblastoma: importance of analytical method

    Energy Technology Data Exchange (ETDEWEB)

    Rundle-Thiele, Dayle [Centre for Clinical Research, University of Queensland, Brisbane, Queensland (Australia); Day, Bryan; Stringer, Brett [Brain Cancer Research Unit, Queensland Institute of Medical Research, Brisbane, Queensland (Australia); Fay, Michael [Department of Radiation Oncology, Royal Brisbane and Women' s Hospital, Brisbane, Queensland (Australia); Martin, Jennifer [Discipline of Clinical Pharmacology, School of Medicine and Public Health, University of Newcastle, Newcastle, New South Wales (Australia); Jeffree, Rosalind L [Department of Neurosurgery, Royal Brisbane and Women' s Hospital, Brisbane, Queensland (Australia); Thomas, Paul [Queensland PET Service, Royal Brisbane and Women' s Hospital, Brisbane, Queensland (Australia); Bell, Christopher [Centre for Clinical Research, University of Queensland, Brisbane, Queensland (Australia); Salvado, Olivier [CSIRO Digital Productivity Flagship, CSIRO, Herston, Queensland (Australia); Gal, Yaniv [Centre for Medical Diagnostic Technologies in Queensland, University of Queensland, Brisbane, Queensland (Australia); Coulthard, Alan [Discipline of Medical Imaging, University of Queensland, St Lucia, Queensland (Australia); Department of Medical Imaging, Royal Brisbane and Women' s Hospital, Brisbane, Queensland (Australia); Crozier, Stuart [Centre for Medical Diagnostic Technologies in Queensland, University of Queensland, Brisbane, Queensland (Australia); Rose, Stephen, E-mail: stephen.rose@csiro.au [CSIRO Digital Productivity Flagship, CSIRO, Herston, Queensland (Australia); Centre for Clinical Research, University of Queensland, Brisbane, Queensland (Australia)

    2015-06-15

    Accurate knowledge of O{sup 6}-methylguanine methyltransferase (MGMT) gene promoter subtype in patients with glioblastoma (GBM) is important for treatment. However, this test is not always available. Pre-operative diffusion MRI (dMRI) can be used to probe tumour biology using the apparent diffusion coefficient (ADC); however, its ability to act as a surrogate to predict MGMT status has shown mixed results. We investigated whether this was due to variations in the method used to analyse ADC. We undertook a retrospective study of 32 patients with GBM who had MGMT status measured. Matching pre-operative MRI data were used to calculate the ADC within contrast enhancing regions of tumour. The relationship between ADC and MGMT was examined using two published ADC methods. A strong trend between a measure of ‘minimum ADC’ and methylation status was seen. An elevated minimum ADC was more likely in the methylated compared to the unmethylated MGMT group (U = 56, P = 0.0561). In contrast, utilising a two-mixture model histogram approach, a significant reduction in mean measure of the ‘low ADC’ component within the histogram was associated with an MGMT promoter methylation subtype (P < 0.0246). This study shows that within the same patient cohort, the method selected to analyse ADC measures has a significant bearing on the use of that metric as a surrogate marker of MGMT status. Thus for dMRI data to be clinically useful, consistent methods of data analysis need to be established prior to establishing any relationship with genetic or epigenetic profiling.

  15. Impact of Margin Status on Local Recurrence After Mastectomy for Ductal Carcinoma In Situ

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    Childs, Stephanie K. [Harvard Radiation Oncology Program, Boston, Massachusetts (United States); Chen, Yu-Hui [Department of Biostatistics and Computational Biology, Dana-Farber Cancer Institute, Boston, Massachusetts (United States); Duggan, Margaret M. [Department of Surgery, Faulkner Hospital, Boston, Massachusetts (United States); Department of Surgery, Brigham and Women' s Hospital and Dana-Farber Cancer Institute, Boston, Massachusetts (United States); Golshan, Mehra [Department of Surgery, Brigham and Women' s Hospital, Boston, Massachusetts (United States); Pochebit, Stephen [Department of Pathology, Faulkner Hospital, Boston, Massachusetts (United States); Punglia, Rinaa S.; Wong, Julia S. [Department of Radiation Oncology, Brigham and Women' s Hospital and Dana-Farber Cancer Institute, Boston, Massachusetts (United States); Bellon, Jennifer R., E-mail: jbellon@lroc.harvard.edu [Department of Radiation Oncology, Brigham and Women' s Hospital and Dana-Farber Cancer Institute, Boston, Massachusetts (United States)

    2013-03-15

    Purpose: To examine the rate of local recurrence according to the margin status for patients with pure ductal carcinoma in situ (DCIS) treated by mastectomy. Methods and Materials: One hundred forty-five consecutive women who underwent mastectomy with or without radiation therapy for DCIS from 1998 to 2005 were included in this retrospective analysis. Only patients with pure DCIS were eligible; patients with microinvasion were excluded. The primary endpoint was local recurrence, defined as recurrence on the chest wall; regional and distant recurrences were secondary endpoints. Outcomes were analyzed according to margin status (positive, close (≤2 mm), or negative), location of the closest margin (superficial, deep, or both), nuclear grade, necrosis, receptor status, type of mastectomy, and receipt of hormonal therapy. Results: The primary cohort consisted of 142 patients who did not receive postmastectomy radiation therapy (PMRT). For those patients, the median follow-up time was 7.6 years (range, 0.6-13.0 years). Twenty-one patients (15%) had a positive margin, and 23 patients (16%) had a close (≤2 mm) margin. The deep margin was close in 14 patients and positive in 6 patients. The superficial margin was close in 13 patients and positive in 19 patients. One patient experienced an isolated invasive chest wall recurrence, and 1 patient had simultaneous chest wall, regional nodal, and distant metastases. The crude rates of chest wall recurrence were 2/142 (1.4%) for all patients, 1/21 (4.8%) for those with positive margins, 1/23 (4.3%) for those with close margins, and 0/98 for patients with negative margins. PMRT was given as part of the initial treatment to 3 patients, 1 of whom had an isolated chest wall recurrence. Conclusions: Mastectomy for pure DCIS resulted in a low rate of local or distant recurrences. Even with positive or close mastectomy margins, the rates of chest wall recurrences were so low that PMRT is likely not warranted.

  16. Coordinate regulation of microenvironmental stimuli and role of methylation in bone metastasis from breast carcinoma.

    Science.gov (United States)

    Matteucci, Emanuela; Maroni, Paola; Disanza, Andrea; Bendinelli, Paola; Desiderio, Maria Alfonsina

    2016-01-01

    The pathogenesis of bone metastasis is unclear, and much focus in metastatic biology and therapy relays on epigenetic alterations. Since DNA-methyltransferase blockade with 5-aza-2'-deoxycytidine (dAza) counteracts tumour growth, here we utilized dAza to clarify whether molecular events undergoing epigenetic control were critical for bone metastatization. In particular, we investigated the patterns of secreted-protein acidic and rich in cysteine (SPARC) and of Endothelin 1, affected by DNA methyltransferases in tumours, with the hypothesis that in bone metastasis a coordinate function of SPARC and Endothelin 1, if any occurs, was orchestrated by DNA methylation. To this purpose, we prepared a xenograft model with the clone 1833, derived from human-MDA-MB231 cells, and dAza administration slowed-down metastasis outgrowth. This seemed consequent to the reductions of SPARC and Endothelin 1 at invasive front and in the bone marrow, mostly due to loss of Twist. In the metastasis bulk Snail, partly reduced by dAza, might sustain Endothelin 1-SPARC cooperativity. Both SPARC and Endothelin 1 underwent post-translational control by miRNAs, a molecular mechanism that might explain the in vivo data. Ectopic miR29a reduced SPARC expression also under long-term dAza exposure, while Endothelin 1 down-regulation occurred in the presence of endogenous-miR98 expression. Notably, dAza effects differed depending on in vivo and in vitro conditions. In 1833 cells exposed to 30-days dAza, SPARC-protein level was practically unaffected, while Endothelin 1 induction depended on the 3'-UTR functionality. The blockade of methyltransferases leading to SPARC reduction in vivo, might represent a promising strategy to hamper early steps of the metastatic process affecting the osteogenic niche.

  17. Ideal number of biopsy tumor fragments for predicting HER2 status in gastric carcinoma resection specimens.

    Science.gov (United States)

    Ahn, Sangjeong; Ahn, Soomin; Van Vrancken, Michael; Lee, Minju; Ha, Sang Yun; Lee, Hyuk; Min, Byung-Hoon; Lee, Jun Haeng; Kim, Jae J; Choi, Sunkyu; Jung, Sin-Ho; Choi, Min Gew; Lee, Jun-Ho; Sohn, Tae Sung; Bae, Jae Moon; Kim, Sung; Kim, Kyoung-Mee

    2015-11-10

    Intratumoral heterogeneity of HER2 expression is common in gastric cancers and pose a challenge for identifying patients who would benefit from anti-HER2 therapy. The aim of this study is to compare HER2 expression in biopsy and resection specimens of gastric carcinoma by immunohistochemistry (IHC) and to find the ideal number of biopsy tumor fragments that can accurately predict HER2 overexpression in the corresponding surgically resected specimen. The HER2 IHC results of 702 paired biopsy and resection specimens of gastric cancer were compared.The mean number of biopsy fragments among all cases was 4.3 (range 1-11). HER2 was positive in 130 (18.5%) endoscopic biopsies and in 102 (14.5%) gastrectomy specimens. Intratumoral heterogeneity of HER2 was found in 80 (61.5%) biopsies and 70 (68.6%) resection specimens. Out of the 70 surgical specimens with intratumoral heterogeneity, 24 (34.3%) of the corresponding biopsies were categorized as negative (positive conversion). In the 86 (12.3%) discrepant cases, negative conversion was observed in 57 (66.3%) cases and positive conversion in 29 (33.7%). The fragment numbers were significantly correlated with the discrepancy of results and positive predictability (P = 0.0315 and P = 0.0052). ROC curve analysis and positive predictability showed that 4 fragments should be obtained to minimize the differences in HER2 scores between biopsy and resection specimen.In gastric carcinomas with discrepant HER2 results between biopsy and surgical resection specimens, intratumoral heterogeneity is common with most of them showing positive conversion. To predict HER2 status precisely, at least 4 biopsy fragments containing tumor cells are required.

  18. XRCC1 Arg194Trp and Arg399Gln polymorphisms and arsenic methylation capacity are associated with urothelial carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Chiang, Chien-I [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Huang, Ya-Li [Department of Public Health, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan (China); Chen, Wei-Jen [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Shiue, Horng-Sheng [Department of Chinese Medicine, Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Taoyuan, Taiwan (China); Huang, Chao-Yuan; Pu, Yeong-Shiau [Department of Urology, National Taiwan University Hospital, College of Medicine National Taiwan University, Taipei, Taiwan (China); Lin, Ying-Chin [Department of Family Medicine, Shung Ho Hospital, Taipei Medical University, New Taipei, Taiwan (China); Department of Health Examination, Wan Fang Hospital, Taipei Medical University, Taipei, Taiwan (China); Division of Family Medicine, School of Medicine, Taipei Medical University, Taipei, Taiwan (China); Hsueh, Yu-Mei, E-mail: ymhsueh@tmu.edu.tw [School of Public Health, College of Public Health and Nutrition, Taipei Medical University, Taipei, Taiwan (China); Department of Public Health, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan (China)

    2014-09-15

    The association between DNA repair gene polymorphisms and bladder cancer has been widely studied. However, few studies have examined the correlation between urothelial carcinoma (UC) and arsenic or its metabolites. The aim of this study was to examine the association between polymorphisms of the DNA repair genes, XRCC1 Arg194Trp, XRCC1 Arg399Gln, XRCC3 Thr241Met, and XPD Lys751Gln, with urinary arsenic profiles and UC. To this end, we conducted a hospital-based case–control study with 324 UC patients and 647 age- and gender-matched non-cancer controls. Genomic DNA was used to examine the genotype of XRCC1 Arg194Trp, XRCC1 Arg399Gln, XRCC3 Thr241Met, and XPD Lys751Gln by PCR-restriction fragment length polymorphism analysis (PCR-RFLP). Urinary arsenic profiles were measured by high performance liquid chromatography (HPLC) linked with hydride generator and atomic absorption spectrometry. The XRCC1 399 Gln/Gln and 194 Arg/Trp and Trp/Trp genotypes were significantly related to UC, and the odds ratio (OR) and 95% confidence interval (95%CI) were 1.68 (1.03–2.75) and 0.66 (0.48–0.90), respectively. Participants with higher total urinary arsenic levels, a higher percentage of inorganic arsenic (InAs%) and a lower percentage of dimethylarsinic acid (DMA%) had a higher OR of UC. Participants carrying XRCC1 risk diplotypes G-C/G-C, A-C/A-C, and A-T/G-T, and who had higher total arsenic levels, higher InAs%, or lower DMA% compared to those with other XRCC1 diplotypes had a higher OR of UC. Our results suggest that the XRCC1 399 Gln/Gln and 194 Arg/Arg DNA repair genes play an important role in poor arsenic methylation capacity, thereby increasing the risk of UC in non-obvious arsenic exposure areas. - Highlights: • The XRCC1 399Gln/Gln genotype was significantly associated with increased OR of UC. • The XRCC1 194 Arg/Trp and Trp/Trp genotype had a significantly decreased OR of UC. • Combined effect of the XRCC1 genotypes and poor arsenic methylation capacity on

  19. Promoter Methylation Precedes Chromosomal Alterations in Colorectal Cancer Development

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    Sarah Derks

    2006-01-01

    Full Text Available Background: Colorectal cancers are characterized by genetic and epigenetic alterations. This study aimed to explore the timing of promoter methylation and relationship with mutations and chromosomal alterations in colorectal carcinogenesis. Methods: In a series of 47 nonprogressed adenomas, 41 progressed adenomas (malignant polyps, 38 colorectal carcinomas and 18 paired normal tissues, we evaluated promoter methylation status of hMLH1, O6MGMT, APC, p14ARF, p16INK4A, RASSF1A, GATA-4, GATA-5, and CHFR using methylation-specific PCR. Mutation status of TP53, APC and KRAS were studied by p53 immunohistochemistry and sequencing of the APC and KRAS mutation cluster regions. Chromosomal alterations were evaluated by comparative genomic hybridization. Results: Our data demonstrate that nonprogressed adenomas, progressed adenomas and carcinomas show similar frequencies of promoter methylation for the majority of the genes. Normal tissues showed significantly lower frequencies of promoter methylation of APC, p16INK4A, GATA-4, and GATA-5 (P-values: 0.02, 0.02, 1.1×10−5 and 0.008 respectively. P53 immunopositivity and chromosomal abnormalities occur predominantly in carcinomas (P values: 1.1×10−5 and 4.1×10−10. Conclusions: Since promoter methylation was already present in nonprogressed adenomas without chromosomal alterations, we conclude that promoter methylation can be regarded as an early event preceding TP53 mutation and chromosomal abnormalities in colorectal cancer development.

  20. The impact of age, marital status, and physician-patient interactions on the care of older women with breast carcinoma.

    Science.gov (United States)

    Silliman, R A; Troyan, S L; Guadagnoli, E; Kaplan, S H; Greenfield, S

    1997-10-01

    Understanding why older women with breast carcinoma do not receive definitive treatment is critical if disparities in mortality between younger and older women are to be reduced. With this in mind, the authors studied 302 women age > or =55 years with early stage breast carcinoma. Data were collected from surgical records and in telephone interviews with the women. The main outcome was receipt of definitive primary tumor therapy, defined either as modified radical mastectomy or as breast-conserving surgery with axillary dissection followed by radiation therapy. The majority (56%) of the women underwent breast-conserving surgery and axillary dissection followed by radiation therapy. After statistical control for four variables (comorbidity, physical function, tumor size, and lymph node status), patients' ages, marital status, and the number of times breast carcinoma specialists discussed treatment options were significantly associated with the receipt of definitive primary tumor therapy. The authors concluded that when older women have been newly diagnosed with breast carcinoma and there is clinical uncertainty as to the most appropriate therapies, patients may be better served if they are offered choices from among definitive therapies. In discussing therapies with them, physicians must be sensitive to their fears and concerns about the monetary costs and functional consequences of treatment in relation to the expected benefits.

  1. CpG promoter methylation status is not a prognostic indicator of gene expression in beryllium challenge.

    Science.gov (United States)

    Tooker, Brian C; Ozawa, Katherine; Newman, Lee S

    2016-05-01

    Individuals exposed to beryllium (Be) may develop Be sensitization (BeS) and progress to chronic beryllium disease (CBD). Recent studies with other metal antigens suggest epigenetic mechanisms may be involved in inflammatory disease processes, including granulomatous lung disorders and that a number of metal cations alter gene methylation. The objective of this study was to determine if Be can exert an epigenetic effect on gene expression by altering methylation in the promoter region of specific genes known to be involved in Be antigen-mediated gene expression. To investigate this objective, three macrophage tumor mouse cell lines known to differentially produce tumor necrosis factor (TNF)-α, but not interferon (IFN)-γ, in response to Be antigen were cultured with Be or controls. Following challenges, ELISA were performed to quantify induced TNFα and IFNγ expression. Bisulfate-converted DNA was evaluated by pyrosequencing to quantify CpG methylation within the promoters of TNFα and IFNγ. Be-challenged H36.12J cells expressed higher levels of TNFα compared to either H36.12E cells or P388D.1 cells. However, there were no variations in TNFα promoter CpG methylation levels between cell lines at the six CpG sites tested. H36.12J cell TNFα expression was shown to be metal-specific by the induction of significantly more TNFα when exposed to Be than when exposed to aluminum sulfate, or nickel (II) chloride, but not when exposed to cobalt (II) chloride. However, H36.12J cell methylation levels at the six CpG sites examined in the TNFα promoter did not correlate with cytokine expression differences. Nonetheless, all three cell lines had significantly more promoter methylation at the six CpG sites investigated within the IFNγ promoter (a gene that is not expressed) when compared to the six CpG sites investigated in the TNFα promoter, regardless of treatment condition (p beryllium had no impact on promoter methylation status, despite its ability to induce pro

  2. THE DIAGNOSTIC ACCURACY OF CHEST CT IN THE DETECTION OF TUMOR AND NODAL STATUS IN NON SMALL CELL LUNG CARCINOMA

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    Aziza Icksan

    2003-12-01

    Full Text Available At this time there is an increasing demand for an accurate preoperative staging in non small cell lung cancer. Chest Computed Tomography (CT is one of the imaging modality of choice used for this purpose. This study evaluated the accuracy of the chest CT to determine the status of the tumor and nodules in non small cell lung cancer. During the years 1998 and 1999, a descriptive prospective study of 32 patients undergoing a contrast enhanced chest CT examination for non small cell lung cancer, stage I-IIIA, was conducted. Lobectomy, lymph nodes dissection and postoperative histo-pathological examination were done. CT findings were as follows: a sensitivity of 100%, a specificity of 25% and an accuracy of 60% in the detection of the nodule stage were found. In 17 patients with adeno-carcinoma, the sensitivity, the specificity and the accuracy were 86.6%, 100% and 88.2% respectively. The diagnosis of all patients was confirmed histo-pathologically. Six patients with T2 and 26 patients with T3 were detected by chest CT; the accuracy of the tumor status was 93.7%, confirmed by surgical and histo-pathological examinations. It was concluded that the CT played an important role in determining the clinical stage of non small cell lung cancer. The specificity and accuracy were higher in adeno-carcinoma as compared with squamous cell carcinoma in detecting the nodal status.

  3. PIN1 in hepatocellular carcinoma is associated with TP53 gene status.

    Science.gov (United States)

    Bae, Jun Sang; Noh, Sang Jae; Kim, Kyoung Min; Jang, Kyu Yun; Park, Ho Sung; Chung, Myoung Ja; Park, Byung-Hyun; Moon, Woo Sung

    2016-10-01

    Phosphorylation of proteins on serine/threonine residues that precede proline (pSer/Thr-Pro) is specifically catalyzed by the peptidyl-prolyl cis-trans isomerase PIN1. PIN1-mediated prolyl-isomerization induces cell cycle arrest and growth inhibition through the regulation of target proteins, including TP53. We examined whether PIN1 acts in a different manner according to TP53 gene status in hepatocellular carcinoma (HCC). We investigated the expression of PIN1 and TP53 proteins in 119 HCC tissue samples. We also analyzed PIN1 expression in combination with TP53 gene mutation and its correlation with the clinical outcome. In addition, we used synthetic small interfering RNA to silence PIN1 gene expression in TP53 wild-type and TP53 mutant HCC cell lines, and then evaluated cell proliferation, migration and invasion. Expression of PIN1 was strongly associated with expression of TP53 protein or TP53 mutation of HCC samples. PIN1 and TP53 expression in TP53 mutant HCC cell lines was higher than that in TP53 wild-type HCC cell lines. Silencing of PIN1 in HLE cells containing mutant TP53 significantly decreased cell proliferation, migration and invasion. In contrast to PIN1 silencing in HLE cells, PIN1 silencing in HepG2 cells containing functional wild-type TP53 resulted in enhanced tumor cell proliferation. HCC patients bearing PIN1 expression with wild-type TP53 were predicted to demonstrate favorable relapse-free survival. Our results suggest that PIN1 plays a role in cancer cell proliferation, migration and invasion in a different manner according to the TP53 gene mutation status in HCC. In particular, interaction of PIN1 with mutant TP53 can act as a tumor promoter and increase its oncogenic activities in HCC.

  4. p53 gene in treatment of hepatic carcinoma:Status quo

    Institute of Scientific and Technical Information of China (English)

    Yong-Song Guan; Zi La; Lin Yang; Qing He; Ping Li

    2007-01-01

    Hepatocellular carcinoma(HCC)is one of the 10 most common cancers worldwide.There is no ideal treatment for HCC yet and many researchers are trying to improve the effects of treatment by changing therapeutic strategies.As the majority of human cancers seem to exhibit either abnormal p53 gene or disrupted p53 gene activation pathways,intervention to restore wild-type p53 (wt-p53)activities is an attractive anti-cancer therapy including HCC.Abnormalities of p53 are also considered a predisposition factor for hepatocarcinogenesis.p53 is frequently mutated in HCC.Most HCCs have defects in the p53-mediated apoptotic pathway although they carry wt-p53.High expression of p53 in vivo may exert therapeutic effects on HCC in two aspects:(1)High expression of exogenous p53 protein induces apoptosis of tumor cells by inhibiting proliferation of cells through several biologic pathways and(2)Exogenous p53 renders HCC more sensitive to some chemotherapeutic agents.Several approaches have been designed for the treatment of HCC via the p53 pathway by restoring the tumor suppression function from inactivation,rescuing the mutated p53 gene from instability,or delivering therapeutic exogenous p53.Products with p53 status as the target have been studied extensively in vitro and in vivo.This review elaborates some therapeutic mechanisms and advances in using recombinant human adenovirus p53 and oncolytic virus products for the treatment of HCC.

  5. Socioeconomic status and esophageal squamous cell carcinoma risk in Kashmir, India.

    Science.gov (United States)

    Dar, Nazir A; Shah, Idrees A; Bhat, Gulzar A; Makhdoomi, Muzamil A; Iqbal, Beenish; Rafiq, Rumaisa; Nisar, Iqra; Bhat, Arshid B; Nabi, Sumaiya; Masood, Akbar; Shah, Sajad A; Lone, Mohd M; Zargar, Showkat A; Islami, Farhad; Boffetta, Paolo

    2013-09-01

    Studies have persistently associated esophageal squamous cell carcinoma (ESCC) risk with low socioeconomic status (SES), but this association is unexplored in Kashmir, an area with a high incidence of ESCC in the northernmost part of India. We carried out a case-control study to assess the association of multiple indicators of SES and ESCC risk in the Kashmir valley. A total number of 703 histologically confirmed ESCC cases and 1664 controls matched to the cases for age, sex, and district of residence were recruited from October 2008 to January 2012. Conditional logistic regression models were used to calculate unadjusted and adjusted odds ratios and 95% confidence intervals. Composite wealth scores were constructed based on the ownership of several appliances using multiple correspondence analyses. Higher education, living in a kiln brick or concrete house, use of liquefied petroleum gas and electricity for cooking, and higher wealth scores all showed an inverse association with ESCC risk. Compared to farmers, individuals who had government jobs or worked in the business sector were at lower risk of ESCC, but this association disappeared in fully adjusted models. Occupational strenuous physical activity was strongly associated with ESCC risk. In summary, we found a strong relationship of low SES and ESCC in Kashmir. The findings need to be studied further to understand the mechanisms through which such SES parameters increase ESCC risk.

  6. Coenzyme Q10 and Oxidative Stress: Inflammation Status in Hepatocellular Carcinoma Patients after Surgery

    Science.gov (United States)

    Liu, Hsiao-Tien; Cheng, Shao-Bin; Huang, Yi-Chia; Huang, Yin-Tzu; Lin, Ping-Ting

    2017-01-01

    (1) Background: Hepatocellular carcinoma (HCC) is the second leading cause of cancer deaths worldwide, and surgical resection is the main treatment for HCC. To date, no published study has examined the status of coenzyme Q10 in patients with HCC after surgery. Thus, the purpose of this study was to investigate the correlations between the level of coenzyme Q10, oxidative stress, and inflammation in patients with HCC after surgery; (2) Methods: 71 primary HCC patients were recruited. Levels of coenzyme Q10, vitamin E, oxidative stress (malondialdehyde), antioxidant enzymes activity (superoxidase dismutase, catalase, and glutathione peroxidase), and inflammatory markers (high sensitivity C-reactive protein; tumor necrosis factor-α; and interleukin-6) were measured; (3) Results: Patients with HCC had a significantly lower levels of coenzyme Q10 (p = 0.01) and oxidative stress (p < 0.01), and significantly higher levels of antioxidant enzymes activities and inflammation after surgery (p < 0.05). The level of coenzyme Q10 was significantly positively correlated with antioxidant capacity (vitamin E and glutathione peroxidase activity) and negatively correlated with inflammation markers after surgery; (4) Conclusion: Hepatocarcinogenesis is associated with oxidative stress, and coenzyme Q10 may be considered an antioxidant therapy for patients with HCC, particularly those with higher inflammation after surgery. PMID:28054958

  7. Abnormal Methylation Status of the GNAS Exon 1A Region in Pseudohypohyperparathyroidism Combined With Turner Syndrome.

    Science.gov (United States)

    Zhu, Jie; Wang, Dong; Ren, An; Xing, Yan; Zhang, Dongliang; Wei, Jun; Yu, Ning; Xing, Xuenong; Ye, Shandong

    2015-12-01

    Pseudohypohyperparathyroidism (PHHP) is a rare type of pseudohypoparathyroidism (PHP), which seems to have a normal skeletal response to parathyroid hormone but shows renal resistance. Almost all patients with PHHP have PHP Ib, a subtype of PHP that is usually caused by GNAS methylation defects, often in exon 1A. Some features of Albright hereditary osteodystrophy can occasionally be found in patients with PHHP, but these features are also common in Turner syndrome. The authors report on an extremely rare case of a patient with PHHP and Turner syndrome, a 47-year-old woman who sought medical attention for hypocalcemia and elevated parathyroid hormone. She had no family history of hypocalcemia and no STX16 gene deletions. She had a mosaic karyotype of 46, X, del(X)(p11.4)/45, XO. Pyrosequencing was performed to determine the GNAS exon 1A methylation. The degree of methylation found in exon 1A of the patient was lower than her unaffected relatives.

  8. Novel Candidate Key Drivers in the Integrative Network of Genes, MicroRNAs, Methylations, and Copy Number Variations in Squamous Cell Lung Carcinoma

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    Tao Huang

    2015-01-01

    Full Text Available The mechanisms of lung cancer are highly complex. Not only mRNA gene expression but also microRNAs, DNA methylation, and copy number variation (CNV play roles in tumorigenesis. It is difficult to incorporate so much information into a single model that can comprehensively reflect all these lung cancer mechanisms. In this study, we analyzed the 129 TCGA (The Cancer Genome Atlas squamous cell lung carcinoma samples with gene expression, microRNA expression, DNA methylation, and CNV data. First, we used variance inflation factor (VIF regression to build the whole genome integrative network. Then, we isolated the lung cancer subnetwork by identifying the known lung cancer genes and their direct regulators. This subnetwork was refined by the Bayesian method, and the directed regulations among mRNA genes, microRNAs, methylations, and CNVs were obtained. The novel candidate key drivers in this refined subnetwork, such as the methylation of ARHGDIB and HOXD3, microRNA let-7a and miR-31, and the CNV of AGAP2, were identified and analyzed. On three large public available lung cancer datasets, the key drivers ARHGDIB and HOXD3 demonstrated significant associations with the overall survival of lung cancer patients. Our results provide new insights into lung cancer mechanisms.

  9. Histological, Immunohistological, and Clinical Features of Merkel Cell Carcinoma in Correlation to Merkel Cell Polyomavirus Status

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    T. Jaeger

    2012-01-01

    Full Text Available Merkel cell carcinoma is a rare, but highly malignant tumor of the skin with high rates of metastasis and poor survival. Its incidence rate rises and is currently about 0.6/100000/year. Clinical differential diagnoses include basal cell carcinoma, cyst, amelanotic melanoma, lymphoma and atypical fibroxanthoma. In this review article clinical, histopathological and immunhistochemical features of Merkel cell carcinoma are reported. In addition, the role of Merkel cell polyomavirus is discussed.

  10. Differential differences in methylation status of putative imprinted genes among cloned swine genomes.

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    Chih-Jie Shen

    Full Text Available DNA methylation is a major epigenetic modification in the mammalian genome that regulates crucial aspects of gene function. Mammalian cloning by somatic cell nuclear transfer (SCNT often results in gestational or neonatal failure with only a small proportion of manipulated embryos producing live births. Many of the embryos that survive to term later succumb to a variety of abnormalities that are likely due to inappropriate epigenetic reprogramming. Aberrant methylation patterns of imprinted genes in cloned cattle and mice have been elucidated, but few reports have analyzed the cloned pig genome. Four surviving cloned sows that were created by ear fibroblast nuclear transfer, each with a different life span and multiple organ defects, such as heart defects and bone growth delay, were used as epigenetic study materials. First, we identified four putative differential methylation regions (DMR of imprinted genes in the wild-type pig genome, including two maternally imprinted loci (INS and IGF2 and two paternally imprinted loci (H19 and IGF2R. Aberrant DNA methylation, either hypermethylation or hypomethylation, commonly appeared in H19 (45% of imprinted loci hypermethylated vs. 30% hypomethylated, IGF2 (40% vs. 0%, INS (50% vs. 5%, and IGF2R (15% vs. 45% in multiple tissues from these four cloned sows compared with wild-type pigs. Our data suggest that aberrant epigenetic modifications occur frequently in the genome of cloned swine. Even with successful production of cloned swine that avoid prenatal or postnatal death, the perturbation of methylation in imprinted genes still exists, which may be one of reason for their adult pathologies and short life. Understanding the aberrant pattern of gene imprinting would permit improvements in future cloning techniques.

  11. Paclitaxel/Taxol sensitivity in human renal cell carcinoma is not determined by the p53 status.

    Science.gov (United States)

    Reinecke, Petra; Kalinski, Thomas; Mahotka, Csaba; Schmitz, Michael; Déjosez, Marion; Gabbert, Helmut Erich; Gerharz, Claus Dieter

    2005-05-26

    In this study, we analyzed the role of the p53 status for paclitaxel/Taxol sensitivity in renal cell carcinomas (RCCs) of the clear cell type. Using immunohistochemistry, nuclear p53 accumulation could not be correlated to the paclitaxel/Taxol sensitivity. DNA sequencing detected a p53 gene mutation in two out of eight RCC cell lines, i.e. in exon 8 (cell line clearCa-6), and in exon 9 (cell line clearCa-5). No correlation, however, was found between the p53 status of our RCC cell lines and their paclitaxel/Taxol sensitivity as indicated by the IC50 values. However, paclitaxel-induced growth inhibition in paclitaxel-sensitive RCC cell lines was accompanied by an increase in apoptosis, irrespective of their p53 status. Although CD95 up-regulation was observed in renal cell carcinoma with wild-type p53 upon paclitaxel treatment, paclitaxel-induced apoptosis itself is triggered independently from the CD95 system. In conclusion, the p53 status cannot predict paclitaxel/Taxol sensitivity in RCC cell lines of the clear cell type.

  12. Gene methylation profiles of normal mucosa, and benign and malignant colorectal tumors identify early onset markers

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    Vatn Morten

    2008-12-01

    Full Text Available Abstract Background Multiple epigenetic and genetic changes have been reported in colorectal tumors, but few of these have clinical impact. This study aims to pinpoint epigenetic markers that can discriminate between non-malignant and malignant tissue from the large bowel, i.e. markers with diagnostic potential. The methylation status of eleven genes (ADAMTS1, CDKN2A, CRABP1, HOXA9, MAL, MGMT, MLH1, NR3C1, PTEN, RUNX3, and SCGB3A1 was determined in 154 tissue samples including normal mucosa, adenomas, and carcinomas of the colorectum. The gene-specific and widespread methylation status among the carcinomas was related to patient gender and age, and microsatellite instability status. Possible CIMP tumors were identified by comparing the methylation profile with microsatellite instability (MSI, BRAF-, KRAS-, and TP53 mutation status. Results The mean number of methylated genes per sample was 0.4 in normal colon mucosa from tumor-free individuals, 1.2 in mucosa from cancerous bowels, 2.2 in adenomas, and 3.9 in carcinomas. Widespread methylation was found in both adenomas and carcinomas. The promoters of ADAMTS1, MAL, and MGMT were frequently methylated in benign samples as well as in malignant tumors, independent of microsatellite instability. In contrast, normal mucosa samples taken from bowels without tumor were rarely methylated for the same genes. Hypermethylated CRABP1, MLH1, NR3C1, RUNX3, and SCGB3A1 were shown to be identifiers of carcinomas with microsatellite instability. In agreement with the CIMP concept, MSI and mutated BRAF were associated with samples harboring hypermethylation of several target genes. Conclusion Methylated ADAMTS1, MGMT, and MAL are suitable as markers for early tumor detection.

  13. Constitutional and somatic methylation status of DMRH19 and KvDMR in Wilms tumor patients

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    Leila C.A. Cardoso

    2012-01-01

    Full Text Available The most frequent epigenetic alterations in Wilms tumor (WT occur at WT2, assigned to 11p15. WT2 consists of two domains: telomeric domain 1 (DMRH19 that contains the IGF2 gene and an imprinted maternally expressed transcript (H19 and centromeric domain 2 (KvDMR that contains the genes KCNQ1, KCNQ1OT1 and CDKN1C. In this work, we used pyrosequencing and MS-MLPA to compare the methylation patterns of DMRH19/KvDMR in blood and tumor samples from 40 WT patients. Normal constitutional KvDMR methylation indicated that most of the epigenetic alterations in WT occur at DMRH19. Constitutional DMRH19 hypermethylation (HM DMRH19 was observed in two patients with Beckwith-Wiedemann syndrome. Pyrosequencing and MS-MLPA showed HM DMRH19 in 28/34 tumor samples: 16/34 with isolated HM DMRH19 and 12/34 with concomitant HM DMRH19 and KvDMR hypomethylation, indicating paternal uniparental disomy. With the exception of one blood sample, the MS-MLPA and pyrosequencing findings were concordant. Diffuse or focal anaplasia was present in five tumor samples and was associated with isolated somatic HM DMRH19 in four of them. Constitutional 11p15 methylation abnormalities were present in 5% of the samples and somatic abnormalities in the majority of tumors. Combined analysis of DMRH19/KvDMR by pyrosequencing and MS-MLPA is beneficial for characterizing epigenetic anomalies in WT, and MS-MLPA is useful and reliable for estimation of DNA methylation in a clinical setting.

  14. Endometrial tumour BRAF mutations and MLH1 promoter methylation as predictors of germline mismatch repair gene mutation status: a literature review.

    Science.gov (United States)

    Metcalf, Alexander M; Spurdle, Amanda B

    2014-03-01

    Colorectal cancer (CRC) that displays high microsatellite instability (MSI-H) can be caused by either germline mutations in mismatch repair (MMR) genes, or non-inherited transcriptional silencing of the MLH1 promoter. A correlation between MLH1 promoter methylation, specifically the 'C' region, and BRAF V600E status has been reported in CRC studies. Germline MMR mutations also greatly increase risk of endometrial cancer (EC), but no systematic review has been undertaken to determine if these tumour markers may be useful predictors of MMR mutation status in EC patients. Endometrial cancer cohorts meeting review inclusion criteria encompassed 2675 tumours from 20 studies for BRAF V600E, and 447 tumours from 11 studies for MLH1 methylation testing. BRAF V600E mutations were reported in 4/2675 (0.1%) endometrial tumours of unknown MMR mutation status, and there were 7/823 (0.9%) total sequence variants in exon 11 and 27/1012 (2.7%) in exon 15. Promoter MLH1 methylation was not observed in tumours from 32 MLH1 mutation carriers, or for 13 MSH2 or MSH6 mutation carriers. MMR mutation-negative individuals with tumour MLH1 and PMS2 IHC loss displayed MLH1 methylation in 48/51 (94%) of tumours. We have also detailed specific examples that show the importance of MLH1 promoter region, assay design, and quantification of methylation. This review shows that BRAF mutations occurs so infrequently in endometrial tumours they can be discounted as a useful marker for predicting MMR-negative mutation status, and further studies of endometrial cohorts with known MMR mutation status are necessary to quantify the utility of tumour MLH1 promoter methylation as a marker of negative germline MMR mutation status in EC patients.

  15. Serum transforming growth factor-β1 level reflects disease status in patients with esophageal carcinoma after radiotherapy

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    AIM: To evaluate the relationship between changes in serum transforming growth factor β1 (TGFβ1) level and curative effect of radiotherapy (RT) in patients with esophageal carcinoma.METHODS: Ninety patients with histologically confirmed esophageal carcinoma were enrolled. Serum samples for TGFβ1 analysis were obtained before and at the end of RT. An enzyme-linked immunosorbent assay was used to measure serum TGFβ1 level. Multivariate analysis was performed to investigate the relationship between disease status and changes in serum TGFβ1 level.RESULTS: Serum TGFβ1 level in patients with esophageal carcinoma before RT was significantly higher than that in healthy controls (P < 0.001). At the end of RT, serum TGFβ1 level was decreased in 67.82% (59/87) of the patients. The overall survival rate at 1,3 and 5 years was 48.28% (42/87), 19.54% (17/87)and 12.64% (11/87), respectively. Main causes of death were local failure and regional lymph node metastasis.In patients whose serum TGFβ1 level decreased after RT,the survival rate at 1, 3 and 5 years was 61.02% (36/59),28.81% (17/59) and 18.64% (11/59), respectively. The survival rate at 1 year was 17.86% (5/28) in patients whose serum TGFβ1 level increased after RT, and all died within 18 mo (P < 0.01).CONCLUSION: Serum TGFβ1 level may be a useful marker for monitoring disease status after RT in patients with esophageal carcinoma.

  16. Propofol effectively inhibits lithium-pilocarpine-induced status epilepticus in rats via downregulation of N-methyl-D-aspartate receptor 2B subunit expression

    Institute of Scientific and Technical Information of China (English)

    Henglin Wang; Zhuoqiang Wang; Weidong Mi; Cong Zhao; Yanqin Liu; Yongan Wang; Haipeng Sun

    2012-01-01

    Status epilepticus was induced via intraperitoneal injection of lithium-pilocarpine. The inhibitory ef-fects of propofol on status epilepticus in rats were judged based on observation of behavior, elec-troencephalography and 24-hour survival rate. Propofol (12.5-100 mg/kg) improved status epilep-ticus in a dose-dependent manner, and significantly reduced the number of deaths within 24 hours of lithium-pilocarpine injection. Western blot results showed that, 24 hours after induction of status epilepticus, the levels of N-methyl-D-aspartate receptor 2A and 2B subunits were significantly in-creased in rat cerebral cortex and hippocampus. Propofol at 50 mg/kg significantly suppressed the increase in N-methyl-D-aspartate receptor 2B subunit levels, but not the increase in N-methyl-D-aspartate receptor 2A subunit levels. The results suggest that propofol can effectively inhibit status epilepticus induced by lithium-pilocarpine. This effect may be associated with down-regulation of N-methyl-D-aspartate receptor 2B subunit expression after seizures.

  17. Does promoter methylation of the SLC30A5 (ZnT5) zinc transporter gene contribute to the ageing-related decline in zinc status?

    Science.gov (United States)

    Coneyworth, L J; Mathers, J C; Ford, D

    2009-05-01

    A decline in Zn status with ageing may contribute to the development of frailty, including impaired immune function, and increased incidence of age-related degenerative diseases. This decline may be a result of reduced dietary Zn intake and/or impaired Zn absorption in the gut. The Zn transporter ZnT5 may play a key role in the absorption of dietary Zn. The corresponding gene (SLC30A5) has a CpG island in its promoter region, so could be regulated by epigenetic mechanisms. It is hypothesised that methylation of the SLC30A5 promoter region is increased with age and that a resulting reduction in ZnT5 expression contributes to the decline in Zn status observed with ageing. This hypothesis has been addressed through (1) studies of effects of SLC30A5 promoter methylation on gene expression in vitro and (2) in vivo measurements of the DNA methylation status of this gene domain. It has been established in vitro that methylation of the human SLC30A5 promoter region results in reduced expression of an associated reporter gene. Second, this gene region shows variable levels of methylation in vivo. Correlation between the level of methylation at this locus and age would support the hypothesis that age-related hypermethylation of this region has the potential to modulate dietary Zn absorption. This premise is being investigated by analysis of additional samples from a human adult cohort to test the hypothesis that methylation of the SLC30A5 promoter region contributes to the age-related decline in Zn status.

  18. Simultaneous Analysis of SEPT9 Promoter Methylation Status, Micronuclei Frequency, and Folate-Related Gene Polymorphisms: The Potential for a Novel Blood-Based Colorectal Cancer Biomarker.

    Science.gov (United States)

    Ravegnini, Gloria; Zolezzi Moraga, Juan Manuel; Maffei, Francesca; Musti, Muriel; Zenesini, Corrado; Simeon, Vittorio; Sammarini, Giulia; Festi, Davide; Hrelia, Patrizia; Angelini, Sabrina

    2015-12-01

    One challenge in colorectal cancer (CRC) is identifying novel biomarkers to be introduced in screening programs. The present study investigated the promoter methylation status of the SEPT9 gene in peripheral blood samples of subjects' positive fecal occult blood test (FOBT). In order to add new insights, we investigated the association between SEPT9 promoter methylation and micronuclei frequency, and polymorphisms in the folate-related pathway genes. SEPT9 promoter methylation, micronuclei frequency, and genotypes were evaluated on 74 individuals' FOBT positive. Individuals were subjected to a colonoscopy that provided written informed consent for study participation. SEPT9 promoter methylation status was significantly lower in the CRC group than controls (p = 0.0006). In contrast, the CaCo2 cell-line, analyzed as a tissue specific model of colon adenocarcinoma, showed a significantly higher percentage of SEPT9 promoter methylation compared to the CRC group (p < 0.0001). Linear regression analysis showed an inverse correlation between micronuclei frequency and the decrease in the methylation levels of SEPT9 promoter region among CRC patients (β = -0.926, p = 0.0001). With regard to genotype analysis, we showed the involvement of the DHFR polymorphism (rs70991108) in SEPT9 promoter methylation level in CRC patients only. In particular, the presence of at least one 19 bp del allele significantly correlates with decreased SEPT9 promoter methylation, compared to the 19 bp ins/ins genotype (p = 0.007). While remaining aware of the strengths and limitations of the study, this represents the first evidence of a novel approach for the early detection of CRC, using SEPT9 promoter methylation, micronuclei frequency and genotypes, with the potential to improve CRC risk assessment.

  19. Simultaneous Analysis of SEPT9 Promoter Methylation Status, Micronuclei Frequency, and Folate-Related Gene Polymorphisms: The Potential for a Novel Blood-Based Colorectal Cancer Biomarker

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    Gloria Ravegnini

    2015-12-01

    Full Text Available One challenge in colorectal cancer (CRC is identifying novel biomarkers to be introduced in screening programs. The present study investigated the promoter methylation status of the SEPT9 gene in peripheral blood samples of subjects’ positive fecal occult blood test (FOBT. In order to add new insights, we investigated the association between SEPT9 promoter methylation and micronuclei frequency, and polymorphisms in the folate-related pathway genes. SEPT9 promoter methylation, micronuclei frequency, and genotypes were evaluated on 74 individuals’ FOBT positive. Individuals were subjected to a colonoscopy that provided written informed consent for study participation. SEPT9 promoter methylation status was significantly lower in the CRC group than controls (p = 0.0006. In contrast, the CaCo2 cell-line, analyzed as a tissue specific model of colon adenocarcinoma, showed a significantly higher percentage of SEPT9 promoter methylation compared to the CRC group (p < 0.0001. Linear regression analysis showed an inverse correlation between micronuclei frequency and the decrease in the methylation levels of SEPT9 promoter region among CRC patients (β = −0.926, p = 0.0001. With regard to genotype analysis, we showed the involvement of the DHFR polymorphism (rs70991108 in SEPT9 promoter methylation level in CRC patients only. In particular, the presence of at least one 19 bp del allele significantly correlates with decreased SEPT9 promoter methylation, compared to the 19 bp ins/ins genotype (p = 0.007. While remaining aware of the strengths and limitations of the study, this represents the first evidence of a novel approach for the early detection of CRC, using SEPT9 promoter methylation, micronuclei frequency and genotypes, with the potential to improve CRC risk assessment.

  20. Prognosis of Glioblastoma With Oligodendroglioma Component is Associated With the IDH1 Mutation and MGMT Methylation Status

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    Jae Kyung Myung

    2014-12-01

    Full Text Available Glioblastoma (GBM with oligodendroglioma component (GBMO is a newly described GBM subtype in the 2007 World Health Organization classification. However, its biological and genetic characteristics are largely unknown. We investigated the clinicopathological and molecular features of 34 GBMOs and compared the survival rate of these patients with those of patients with astrocytoma, oligodendroglioma, anaplastic oligoastrocytoma (AOA, and conventional GBMs in our hospital. GBMO could be divided into two groups based on the presence of an IDH1 mutation. The IDH1 mutation was more frequently found in secondary GBMO, which had lower frequencies of EGFR amplification but higher MGMT methylation than the wild type IDH1 group, and patients with mutant IDH1 GBMO were on average younger than those with wild-type IDH1. Therefore, GBMO is a clinically and molecularly heterogeneous subtype, largely belonging to a proneural and classical subtype of GBM. The survival rate of GBMO patients itself was worse than that of AOA patients but not significantly better than that of conventional GBM patients. GBMO survival was independent of the dominant histopathological subtype i.e., astrocyte-dominant or oligodendroglioma -dominant, but it was significantly associated with the IDH1 mutation and MGMT methylation status. Therefore, GBMO should be regarded as a separate entity from AOA and must be classified as a subtype of GBM. However, further study is needed to determine whether it is a pathologic variant or a pattern of GBM because GBMO has a similar prognosis to conventional GBMs.

  1. Sensitivity Analysis of the MGMT-STP27 Model and Impact of Genetic and Epigenetic Context to Predict the MGMT Methylation Status in Gliomas and Other Tumors.

    Science.gov (United States)

    Bady, Pierre; Delorenzi, Mauro; Hegi, Monika E

    2016-05-01

    The methylation status of the O(6)-methylguanine-DNA methyltransferase (MGMT) gene is an important predictive biomarker for benefit from alkylating agent therapy in glioblastoma. Our model MGMT-STP27 allows prediction of the methylation status of the MGMT promoter using data from the Illumina's Human Methylation BeadChips (HM-27K and HM-450K) that is publically available for many cancer data sets. Here, we investigate the impact of the context of genetic and epigenetic alterations and tumor type on the classification and report on technical aspects, such as robustness of cutoff definition and preprocessing of the data. The association between gene copy number variation, predicted MGMT methylation, and MGMT expression revealed a gene dosage effect on MGMT expression in lower grade glioma (World Health Organization grade II/III) that in contrast to glioblastoma usually carry two copies of chromosome 10 on which MGMT resides (10q26.3). This implies some MGMT expression, potentially conferring residual repair function blunting the therapeutic effect of alkylating agents. A sensitivity analyses corroborated the performance of the original cutoff for various optimization criteria and for most data preprocessing methods. Finally, we propose an R package mgmtstp27 that allows prediction of the methylation status of the MGMT promoter and calculation of appropriate confidence and/or prediction intervals. Overall, MGMT-STP27 is a robust model for MGMT classification that is independent of tumor type and is adapted for single sample prediction.

  2. RASSF1A and DOK1 Promoter Methylation Levels in Hepatocellular Carcinoma, Cirrhotic and Non-Cirrhotic Liver, and Correlation with Liver Cancer in Brazilian Patients

    Science.gov (United States)

    Araújo, Oscar C.; Rosa, Agatha S.; Fernandes, Arlete; Niel, Christian; Villela-Nogueira, Cristiane A.; Pannain, Vera; Araujo, Natalia M.

    2016-01-01

    Hepatocellular carcinoma (HCC) is the second most common cause of cancer mortality worldwide. Most cases of HCC are associated with cirrhosis related to chronic hepatitis B virus or hepatitis C virus infections. Hypermethylation of promoter regions is the main epigenetic mechanism of gene silencing and has been involved in HCC development. The aim of this study was to determine whether aberrant methylation of RASSF1A and DOK1 gene promoters is associated with the progression of liver disease in Brazilian patients. Methylation levels were measured by pyrosequencing in 41 (20 HCC, 9 cirrhotic, and 12 non-cirrhotic) liver tissue samples. Mean rates of methylation in RASSF1A and DOK1 were 16.2% and 12.0% in non-cirrhotic, 26.1% and 19.6% in cirrhotic, and 59.1% and 56.0% in HCC tissues, respectively, showing a gradual increase according to the progression of the disease, with significantly higher levels in tumor tissues. In addition, hypermethylation of RASSF1A and DOK1 was found in the vast majority (88%) of the HCC cases. Interestingly, DOK1 methylation levels in HCC samples were significantly higher in the group of younger (<40 years) patients, and higher in moderately differentiated than in poorly differentiated tumors (p < 0.05). Our results reinforce the hypothesis that hypermethylation of RASSF1A and DOK1 contributes to hepatocarcinogenesis and is associated to clinicopathological characteristics. RASSF1A and DOK1 promoter hypermethylation may be a valuable biomarker for early diagnosis of HCC and a potential molecular target for epigenetic-based therapy. PMID:27078152

  3. Res earch on CpG-island methylation of CDH13 and DAPK genes in patients with non-small cell lung carcinoma%非小细胞肺癌CDH13及DAPK基因CpG岛甲基化的研究

    Institute of Scientific and Technical Information of China (English)

    黄晖; 孔勇; 冯旭; 彭朝阳

    2015-01-01

    Objective To explore CpG island methylation and clinical significance of CDH13 and DAPK genes in non-small cell lung carcinoma patients.Methods Methylation specific PCR was used to detect the CpG island methylation status of CDH13 and DAPK genes among 40 patients with non-small cell lung carcinoma(lung cancer group) and 30 patients with benign lung diseases(benign lung disease group)R.esults The incidences of CpG island methylation of CDH 13 and DAPK genes were 47.5%(19/40) and 40.0%(16/40) respectively in the lung cancer group,27.5%(11/40) and 30.0%(12/40) respectively in the para-carcinoma group, and 0 in the benign lung disease group.The incidences of CpG island methylation of CDH13 and DAPK genes were significantly higher in the lung cancer group and para-carcinoma group in contrast with those in the benign lung disease group(P0.05).There was no significant difference in the incidences of CpG island methylation of CDH13 and DAPK genes between lung squamous carcinoma tissues and lung adenocarcinoma tissues(P>0.05).Conclusion The CpG island methylation of CDH13and DAPK genes can be observed in the non-small cell lung cancer tissues, and it is associated with the development of non-small cell lung carcinoma.%目的:探讨非小细胞肺癌患者抑癌基因CDH13及DAPK基因CpG岛甲基化情况及其临床意义。方法采用甲基化特异性-PCR方法检测40例非小细胞肺癌患者(肺癌组)和30例肺良性病变患者(肺良性病变组)的抑癌基因CDH13及DAPK的CpG岛甲基化的情况。结果肺癌组织 CDH13、DAPK 基因 CpG 岛甲基化率分别为47.5%(19/40)、40.0%(16/40),癌旁组织分别为27.5%(11/40)、30.0%(12/40),肺良性病变组织均为0。癌组织、癌旁组织CDH13、DAPK的CpG岛甲基化率均明显高于良性病变组(P<0.05),但癌组织与癌旁组织间比较,差异无统计学意义(P>0.05)。鳞癌与腺癌组织CDH13、DAPK的CpG岛甲基化

  4. Restoration of the methylation status of hypermethylated gene promoters by microRNA-29b in human breast cancer: A novel epigenetic therapeutic approach

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    Athena Starlard-Davenport

    2013-01-01

    Full Text Available It is well established that transcriptional silencing of critical tumor-suppressor genes by DNA methylation is a fundamental component in the initiation of breast cancer. However, the involvement of microRNAs (miRNAs in restoring abnormal DNA methylation patterns in breast cancer is not well understood. Therefore, we investigated whether miRNA-29b, due to its complimentarity to the 3′- untranslated region of DNA methyltransferase 3A (DNMT3A and DNMT3B, could restore normal DNA methylation patterns in human breast cancers and breast cancer cell lines. We demonstrated that transfection of pre-miRNA-29b into less aggressive MCF-7 cells, but not MDA-MB-231 mesenchymal cells, inhibited cell proliferation, decreased DNMT3A and DNMT3B messenger RNA (mRNA, and decreased promoter methylation status of ADAM23 , CCNA1, CCND2, CDH1, CDKN1C, CDKN2A, HIC1, RASSF1, SLIT2, TNFRSF10D, and TP73 tumor-suppressor genes. Using methylation polymerase chain reaction (PCR arrays and real-time PCR, we also demonstrated that the methylation status of several critical tumor-suppressor genes increased as stage of breast disease increased, while miRNA-29b mRNA levels were significantly decreased in breast cancers versus normal breast. This increase in methylation status was accompanied by an increase in DNMT1 and DNMT3B mRNA in advanced stage of human breast cancers and in MCF-7, MDA-MB-361, HCC70, Hs-578T, and MDA-MB-231 breast cancer cells as compared to normal breast specimens and MCF-10-2A, a non-tumorigenic breast cell line, respectively. Our findings highlight the potential for a new epigenetic approach in improving breast cancer therapy by targeting DNMT3A and DNMT3B through miRNA-29b in non-invasive epithelial breast cancer cells.

  5. Angiogenesis, Inflammation, Platelets Count, and Metastatic Status as a Predictor for Thrombosis Risk in Nasopharyngeal Carcinoma Patients

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    Aru W Sudoyo

    2015-03-01

    Full Text Available Aim: to assess the use of of angiogenesis, inflammation, platelets count, and metastatic status as predictors for thrombosis risk represented by soluble P-selectin level in nasopharyngeal carcinoma (NPC patients. Methods: a cross sectional study was conducted on NPC patients at the Hematology and Oncology Clinic of Cipto Mangunkusumo Hospital, Jakarta, during Mei to October 2012. Data regarding angiogenesis (CD105 and VEGFR-2, inflammation (IL-6, platelets count, and metastatic status were assessed at enrollment, as well as soluble P-selectin levels in all eligible patients. Bivariate analysis continued with multiple linear regression analysis were done to identify independent predictors for soluble P-selectin levels. Results: sixty NPC patients were enrolled in the study. There was correlation between platelet counts (r=0.389; p=0.002, IL-6 (r=0.595; p<0.001 and number of metastatic sites (r=0.542; p<0.001 with soluble P-selectin level, and a linear regression analysis showed that these three variables can predict soluble P-selectin levels with adjusted R-square 65%. There was no correlation between VEGFR-2 and CD105 levels with soluble P-selectin levels.Conclusion: platelet counts, IL-6 level, and number of sites of metastasis can be used as predictors of soluble P-selectin level as parameter of thrombosis risk in NPC patients. Key words: nasopharyngeal carcinoma (NPC, thrombosis risk, soluble P-selectin.

  6. Association of methylation status of RASSF1A and APC promoters with prostate cancer%RASSF1A和APC基因启动子区甲基化与前列腺癌的关系

    Institute of Scientific and Technical Information of China (English)

    梁永; 董柏萍; 邱镇; 崔心刚; 张连升; 侯建国; 曹广文; 崔飞伦; 张宏伟

    2012-01-01

    目的 研究Ras相关区域家族1A (Ras association domain family l isoform A,RASSF1A)及结肠腺瘤性息肉病(adenomatosis polyposis coli,APC)基因启动子区CpG甲基化及与前列腺癌(PCa)之间的关系,探索PCa早期诊断的检测方法.方法 收集60例PCa和40例前列腺增生(BPH)病例的组织标本及其相关的临床指标.应用亚硫酸氢盐修饰后测序法检测PCa组织及BPH组织中RASSF1A、APC基因启动子区CpG甲基化情况.结果 PCa组RASSF1A、APC基因CG位点甲基化率高于BPH组(60.8% vs 14%,48.84% vs 1.19%,P<0.05);基因甲基化率与PSA、Gleason评分、病理分期和PCa危险分期关系密切(P<0.01); RASSF1A及APC基因甲基化联合检测用于判断PCa及BPH的敏感度和特异度是95.74%和82.9%.结论 RASSFIA、APC基因启动子区甲基化与PCa发生及发展有关,其甲基化率的变化与PCa的危险分期关系密切.检测前列腺组织中相关基因甲基化状态,有望成为诊断早期PCa的一种方法.%Objective To investigate the promoter methylation status of Ras association domain family 1 isoform A (RASSF1A) and adenomatosis polyposis cob' promoter (AFC) and its association with prostate cancer (PCa), so as to pave a way for early diagnosis of Pea. Methods The prostatic tissues and clinical data were collected from 60 patients with prostate cancer and 40 patients with benign prostatic hyperplasia (BPH) tissues. Bisulphite Sequencing PCR was used to detect the promoter CpG methylation of RASSF1A and APC genes. Results The methylation rates of promoter CG sites of RASSF1A and APC genes in Pea group were significantly higher whan those in the BPH group (RASSF1A: 60. 8% vs 14%s APC: 48. 84% vs 1. 19%, P<0. 05). The methylation rates in RASSF1A and APC genes were significantly associated with PSA, Gleason score, pathologic stage and risk classification of PCa (P< 0.01). Combined detection of methylation status of RASSF1A and APC genes yielded a sensitivity of 95. 74

  7. MGMT promoter methylation status and MGMT and CD133 immunohistochemical expression as prognostic markers in glioblastoma patients treated with temozolomide plus radiotherapy

    Directory of Open Access Journals (Sweden)

    Melguizo Consolación

    2012-12-01

    Full Text Available Abstract Background The CD133 antigen is a marker of radio- and chemo-resistant stem cell populations in glioblastoma (GBM. The O6-methylguanine DNA methyltransferase (MGMT enzyme is related with temozolomide (TMZ resistance. Our propose is to analyze the prognostic significance of the CD133 antigen and promoter methylation and protein expression of MGMT in a homogenous group of GBM patients uniformly treated with radiotherapy and TMZ. The possible connection between these GBM markers was also investigated. Methods Seventy-eight patients with GBM treated with radiotherapy combined with concomitant and adjuvant TMZ were analyzed for MGMT and CD133. MGMT gene promoter methylation was determined by methylation-specific polymerase chain reaction after bisulfite treatment. MGMT and CD133 expression was assessed immunohistochemically using an automatic quantification system. Overall and progression-free survival was calculated according to the Kaplan–Meier method. Results The MGMT gene promoter was found to be methylated in 34 patients (44.7% and unmethylated in 42 patients (55.3%. A significant correlation was observed between MGMT promoter methylation and patients’ survival. Among the unmethylated tumors, 52.4% showed low expression of MGMT and 47.6% showed high-expression. Among methylated tumors, 58.8% showed low-expression of MGMT and 41.2% showed high-expression. No correlation was found between MGMT promoter methylation and MGMT expression, or MGMT expression and survival. In contrast with recent results, CD133 expression was not a predictive marker in GBM patients. Analyses of possible correlation between CD133 expression and MGMT protein expression or MGMT promoter methylation were negative. Conclusions Our results support the hypothesis that MGMT promoter methylation status but not MGMT expression may be a predictive biomarker in the treatment of patients with GBM. In addition, CD133 should not be used for prognostic evaluation of these

  8. MicroRNA alterations and associated aberrant DNA methylation patterns across multiple sample types in oral squamous cell carcinoma

    DEFF Research Database (Denmark)

    Wiklund, Erik Digman; Gao, Shan; Hulf, Toby;

    2011-01-01

    MicroRNA (miRNA) expression is broadly altered in cancer, but few studies have investigated miRNA deregulation in oral squamous cell carcinoma (OSCC). Epigenetic mechanisms are involved in the regulation of >30 miRNA genes in a range of tissues, and we aimed to investigate this further in OSCC....

  9. Examination of DNA methylation status of the ELOVL2 marker may be useful for human age prediction in forensic science.

    Science.gov (United States)

    Zbieć-Piekarska, Renata; Spólnicka, Magdalena; Kupiec, Tomasz; Makowska, Żanetta; Spas, Anna; Parys-Proszek, Agnieszka; Kucharczyk, Krzysztof; Płoski, Rafał; Branicki, Wojciech

    2015-01-01

    Age estimation in forensic investigations may complement the prediction of externally visible characteristics and the inference of biogeographical ancestry, thus allowing a better description of an unknown individual. Multiple CpG sites that show linear correlation between age and degree of DNA methylation have been identified in the human genome, providing a selection of candidates for age prediction. In this study, we optimized an assay based on bisulfite conversion and pyrosequencing of 7 CpG sites located in the ELOVL2 gene. Examination of 303 blood samples collected from individuals aged 2-75 years allowed selection of the most informative site, explaining 83% of variation in age. The final linear regression model included two CpG sites in ELOVL2 and enabled age prediction with R(2)=0.859, prediction error=6.85 and mean absolute deviation MAD=5.03. Examination of a testing set of 124 blood samples (MAD=5.75) showed that 68.5% of samples were correctly predicted, assuming that chronological and predicted ages matched ± 7 years. It was found that the ELOVL2 methylation status in bloodstains had not changed significantly after 4 weeks of storage in room temperature conditions. Analysis of 45 bloodstains deposited on tissue paper after 5, 10 and 15 years of storage in room conditions indicated that although a gradual decrease of positive PCR results was observed, the general age prediction success rate remained similar and equaled 60-78%. The obtained results show that the ELOVL2 locus provides a very good source of information about human chronological age based on analysis of blood, including bloodstains, and it may constitute a powerful and reliable predictor in future forensic age estimation models.

  10. O6-Methylguanine-Methyltransferase (MGMT Promoter Methylation Status in Glioma Stem-Like Cells is Correlated to Temozolomide Sensitivity Under Differentiation-Promoting Conditions

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    Lucie Karayan-Tapon

    2012-06-01

    Full Text Available Glioblastoma (GBM is the most malignant type of primary brain tumor with a very poor prognosis. The actual standard protocol of treatment for GBM patients consists of radiotherapy and concomitant temozolomide (TMZ. However, the therapeutic efficacy of this treatment is limited due to tumor recurrence and TMZ resistance. Recently isolated, glioma stem-like cells (GSCs are thought to represent the population of tumorigenic cells responsible for GBM resistance and recurrence following surgery and chemotherapy. In addition, MGMT (O6-methylguanine-methyltransferase methylation is considered as one of the principal mechanisms contributing to TMZ sensitivity of GBM. In this study we have isolated GSCs from 10 adult GBM patients and investigated the relationship between MGMT methylation status and Temozolomide (TMZ sensitivity of these lines grown either in stem-like or differentiation promoting conditions. Sensitivity to TMZ was significantly associated with MGMT methylation status in cells committed to differentiation but not in stem-like cells. In addition, patients harboring highly methylated MGMT promoters had a longer overall survival. These results reveal the importance of the differentiation process when considering the predictive value of MGMT status in GSCs for clinical response to TMZ.

  11. Dietary Chromium Restriction of Pregnant Mice Changes the Methylation Status of Hepatic Genes Involved with Insulin Signaling in Adult Male Offspring

    Science.gov (United States)

    Zhang, Qian; Sun, Xiaofang; Xiao, Xinhua; Zheng, Jia; Li, Ming; Yu, Miao; Ping, Fan; Wang, Zhixin; Qi, Cuijuan; Wang, Tong; Wang, Xiaojing

    2017-01-01

    Maternal undernutrition is linked with an elevated risk of diabetes mellitus in offspring regardless of the postnatal dietary status. This is also found in maternal micro-nutrition deficiency, especial chromium which is a key glucose regulator. We investigated whether maternal chromium restriction contributes to the development of diabetes in offspring by affecting DNA methylation status in liver tissue. After being mated with control males, female weanling 8-week-old C57BL mice were fed a control diet (CON, 1.19 mg chromium/kg diet) or a low chromium diet (LC, 0.14 mg chromium/kg diet) during pregnancy and lactation. After weaning, some offspring were shifted to the other diet (CON-LC, or LC-CON), while others remained on the same diet (CON-CON, or LC-LC) for 29 weeks. Fasting blood glucose, serum insulin, and oral glucose tolerance test was performed to evaluate the glucose metabolism condition. Methylation differences in liver from the LC-CON group and CON-CON groups were studied by using a DNA methylation array. Bisulfite sequencing was carried out to validate the results of the methylation array. Maternal chromium limitation diet increased the body weight, blood glucose, and serum insulin levels. Even when switched to the control diet after weaning, the offspring also showed impaired glucose tolerance and insulin resistance. DNA methylation profiling of the offspring livers revealed 935 differentially methylated genes in livers of the maternal chromium restriction diet group. Pathway analysis identified the insulin signaling pathway was the main process affected by hypermethylated genes. Bisulfite sequencing confirmed that some genes in insulin signaling pathway were hypermethylated in livers of the LC-CON and LC-LC group. Accordingly, the expression of genes in insulin signaling pathway was downregulated. There findings suggest that maternal chromium restriction diet results in glucose intolerance in male offspring through alterations in DNA methylation which

  12. Folic acid supplementation prevents the changes in hepatic promoter methylation status and gene expression in intrauterine growth-retarded piglets during early weaning period.

    Science.gov (United States)

    Jing-Bo, L; Ying, Y; Bing, Y; Xiang-Bing, M; Zhi-Qing, H; Guo-Quan, H; Hong, C; Dai-Wen, C

    2013-10-01

    During intrauterine life, genome interacts with maternal signals to influence the mRNA expression levels of specific genes persistently by regulating DNA methylation status. The objective of this study was to examine the responses of glucocorticoid receptor (GR), peroxisome proliferator-activated receptor alpha and gamma (PPARα and PPARγ) promoter methylation, mRNA expression of genes involved in energy metabolism and metabolite concentrations of intrauterine growth-retarded (IUGR) piglets to dietary folic acid supplementation. According to a 2 × 2 factorial arrangement, 16 IUGR and 16 normal birth weight (NBW) piglets were fed a basal diet or a basal diet supplemented with 5 mg/kg of folic acid from weaning (day 14) to day 35 of age. Triglycerides in hepatic tissue and plasma were significantly elevated in control diets-fed IUGR piglets compared with NBW piglets but were decreased by dietary folic acid supplementation. Hepatic mRNA expression levels of GR, PPARα, PPARγ, fatty acid synthase and phosphoenolpyruvate carboxykinase (PEPCK) in IUGR piglets fed a control diet were significantly higher than that in NBW piglets, and promoter methylation status of GR, PPARα and PPARγ in IUGR piglets was reduced significantly compared with NBW piglets. However, the changes in gene expression and DNA methylation status of IUGR piglets were reversed by dietary folic acid supplementation. Hepatic DNA methyltransferase activity was greater with dietary folic acid supplementation regardless of birth weight. Taken together, these results demonstrated that folic acid supplementation during early period of life could prevent the changes of promoter methylation status and gene expressions in the liver of IUGR piglets.

  13. LINE-1 Methylation Status Correlates Significantly to Post-Therapeutic Recurrence in Stage III Colon Cancer Patients Receiving FOLFOX-4 Adjuvant Chemotherapy.

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    Yun-Ting Lou

    Full Text Available Methylation levels of long interspersed nucleotide elements (LINE-1 are representative of genome-wide methylation status and crucial in maintaining genomic stability and expression. Their prognostic impact on colon cancer patients receiving adjuvant chemotherapy has not been well established. We evaluated the association between LINE-1 methylation status and clinicopathologic features and postoperative oncological outcomes in stage III colon cancer patients.129 UICC stage III colon cancer patients who had received radical resection and FOLFOX adjuvant chemotherapy were enrolled. Global methylation was estimated by analyzing tumor LINE-1 methylation status using bisulfite-polymerase chain reaction (PCR and pyrosequencing assay. Demographics, clinicopathological data, and postoperative outcomes were recorded by trained abstractors. Outcome measurements included postoperative recurrence and disease-free survival. Univariate, multivariate, and survival analyses were conducted to identify prognostic factors of oncological outcomes.The LINE-1 methylation of all 129 patients was measured on a 0-100 scale (mean 63.3; median 63.7, standard deviation 7.1, LINE-1 hypomethylation was more common in patients aged 65 years and above (61.7%±7.6% vs. 64.6±6.4, p=0.019 and those with post-therapeutic recurrence (61.7±7.4 vs 64.3±6.7, p=0.041. Considering risk adjustment, LINE-1 hypomethylation was found to be an independent risk factor of post-therapeutic recurrence (Adjusted OR=14.1, p=0.012. Kaplan-Meier analysis indicated that patients in the low methylation group had shorter period of disease free survival (p=0.01. In a stratified analysis that included 48 patients with post-therapeutic recurrence, it was found that those who experienced shorter period of disease free survival (≦6 months appeared to have lower LINE-1 methylation levels than patients who reported of recurrence after 6 months (56.68±15.75 vs. 63.55±7.57, p=0.041.There was a

  14. Correlation of chromosome damage and promoter methylation status of the DNA repair genes MGMT and hMLH1 in Chinese vinyl chloride monomer (VCM-exposed workers

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    Fen Wu

    2013-02-01

    Full Text Available Objective: To explore the association of the methylation status of MGMT and hMLH1 with chromosome damage induced by vinyl chloride monomer (VCM. Materials and Methods: Methylation of MGMT and hMLH1 was measured in 101 VCM-exposed workers by methylation-specifi c PCR. Chromosome damage in peripheral blood lymphocytes was measured by the cytokinesis-block micronucleus assay. The subjects were divided into chromosome damaged and non-damaged groups based on the normal reference value of micronuclei frequencies determined for two control groups. Results: MGMT promoter methylation was detectable in 5 out of 49 chromosome damaged subjects, but not in the chromosome non-damaged subjects; there was a signifi cant difference in MGMT methylation between the two groups (p < 0.05. Conclusions: We detected aberrant promoter methylation of MGMT in a small number of chromosome damaged VCM-exposed workers, but not in the chromosome non-damaged subjects. This preliminary observation warrants further investigation in a larger study.

  15. Identification of methylation markers for the prediction of nodal metastasis in oral and oropharyngeal squamous cell carcinoma

    NARCIS (Netherlands)

    Melchers, L. J.; Clausen, M. J. A. M.; Mastik, M. F.; Slagter-Menkema, L.; van der Wal, J. E.; Wisman, G. B. A.; Roodenburg, J. L. N.; Schuuring, E.

    2015-01-01

    Hypermethylation is an important mechanism for the dynamic regulation of gene expression, necessary for metastasizing tumour cells. Our aim is to identify methylation tumour markers that have a predictive value for the presence of regional lymph node metastases in patients with oral and oropharyngea

  16. Relationship between Methylation Status of Multi-drug Resistance Protein(MRP) and Multi-drug Resistance in Lung Cancer Cell Lines

    Institute of Scientific and Technical Information of China (English)

    LIU Rui-jun; ZHONG Hong

    2007-01-01

    Objective: To study the relationship between the methylation status of multi-drug resistance protein (MRP) gene and the expression of its mRNA and protein in lung cancer cell lines. Methods: Human embryo lung cell line WI-38, lung adenocarcinoma cell line SPCA-1 and its drug-resistant cells induced by different concentrations of doxorubicin were treated with restriction endonuclease Eco47Ⅲ. The methylation status of MRP was examined by PCR, and the expressions of its mRNA and protein were evaluated by in situ hybridization and immunohistochemistry. Results: MRP gene promoter region of WI-38 cells was in hypermethylation status, but the promoter region of MRP in SPCA-1 cells and their resistant derivatives induced by different concentrations of doxorubicin were in hypomethylation status. There were significant differences in the expression of MRP mRNA among WI-38 cell line, SPCA-1 cells and their drug-resistant derivatives induced by different concentration of doxorubicin. Consistently, MRP immunostaining presented similar significant differences. Conclusion: The promoter region of MRP in SPCA-1 lung adenocarcinoma cells was in hypomethylation status. The hypomethylation status of 5' regulatory region of MRP promoter is an important structural basis that can increase the activity of transcription and results in the development of drug resistance in lung cancer.

  17. Prognostic impact of body mass index stratified by smoking status in patients with esophageal squamous cell carcinoma

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    Sun P

    2016-10-01

    Full Text Available Peng Sun,1,2,* Fei Zhang,1,2,* Cui Chen,3,* Chao Ren,1,2 Xi-Wen Bi,1,2 Hang Yang,1,2 Xin An,1,2 Feng-Hua Wang,1,2 Wen-Qi Jiang1,2 1State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, 2Department of Medical Oncology, Sun Yat-Sen University Cancer Center, 3Department of Oncology, the First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, People’s Republic of China *These authors contributed equally to this work Background: As smoking affects the body mass index (BMI and causes the risk of esophageal squamous cell carcinoma (ESCC, the prognostic impact of BMI in ESCC could be stratified by smoking status. We investigated the true prognostic effect of BMI and its potential modification by smoking status in ESCC. Methods: We retrospectively analyzed 459 patients who underwent curative treatment at a single institution between January 2007 and December 2010. BMI was calculated using the measured height and weight before surgery. Chi-square test was used to evaluate the relationships between smoking status and other clinicopathological variables. The Cox proportional hazard models were used for univariate and multivariate analyses of variables related to overall survival. Results: BMI <18.5 kg/m2 was a significantly independent predictor of poor survival in the overall population and never smokers after adjusting for covariates, but not in ever smokers. Among never smokers, underweight patients (BMI <18.5 kg/m2 had a 2.218 times greater risk of mortality than non-underweight (BMI =18.5 kg/m2 patients (P=0.015. Among ever smokers, BMI <18 kg/m2 increased the risk of mortality to 1.656 (P=0.019, compared to those having BMI =18 kg/m2. Conclusion: Our study is likely the first to show that the prognostic effect of BMI was substantial in ESCC, even after stratifying by smoking status. Furthermore, the risk of death due to low BMI would be significantly increased in never smokers. We believe that

  18. Increased intragenic IGF2 methylation is associated with repression of insulator activity and elevated expression in serous ovarian carcinoma

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    Zhiqing eHuang

    2013-05-01

    Full Text Available Overexpression of insulin-like growth factor-II (IGF2 is a prominent characteristic of many epithelial ovarian malignancies. IGF2 imprinting and transcription are regulated in part through DNA methylation, which in turn regulates binding of the insulator protein, CTCF, within the IGF2/H19 imprint center. We have shown that IGF2 overexpression in ovarian cancer is associated with hypermethylation of CTCF binding sites within the IGF2/H19 imprint center. The aim of this study was to investigate the methylation and binding capacity of a novel putative CTCF binding motif located intragenic to IGF2 and determine how this relates to IGF2 expression. In 35 primary serous epithelial ovarian cancer specimens, methylation of two CpGs, including one within the core binding motif and another adjacent to this motif, was higher in the 18 cancers with elevated IGF2 expression versus 10 with low expression (avg. 68.2% vs. 38.5%; p<0.0001. We also found that the CpG site within the CTCF binding motif is hypermethylated in male gametes (>92%; avg. 93.2%; N=16. We confirmed binding of CTCF to this region in ovarian cancer cells, as well as the paralog of CTCF, BORIS, which is frequently overexpressed in cancers. The unmethylated CTCF binding motif has insulator activity in cells that express CTCF or BORIS, but not in cells that express both CTCF and BORIS. These intragenic CpG dinucleotides comprise a novel paternal germline imprint mark and are located in a binding motif for the insulator protein CTCF. Methylation of the CpG dinucleotides is positively correlated with IGF2 transcription, supporting that increased methylation represses insulator function. These combined results suggest that methylation and CTCF binding at this region play important roles in regulating the level of IGF2 transcription. Our data have revealed a novel epigenetic regulatory element within the IGF2/H19 imprinted domain that is highly relevant to aberrant IGF2 expression in ovarian

  19. [BRAF-STATUS OF PAPILLARY THYROID CARCINOMAS AND STRATEGY OF SURGICAL TREATMENT].

    Science.gov (United States)

    Tarashchenko, Yu N; Kovalenko, A E; Bolgov, M Yu; Guda, B B; Shelkovoy, E A; Nekrasov, K A; Mankovskaya, S; Kashuba, V I

    2015-06-01

    Analyzed the presence of BRAF V600E mutation in the focal thyroid gland in the preoperative diagnosis of papillary carcinoma (PC). Molecular genetic testing conducted on puncture aspirates from 26 patients before surgery. The diagnosis was verified according to the morphological investigations. Mutations in BRAF V600E detected only in patients with the thyroid PC. Thus, the definition of BRAF V600E mutation may be a marker in the preoperative diagnosis of thyroid PC. Analyzed the presence of BRAF V600E mutation in the focal thyroid gland in the preoperative diagnosis of papillary carcinoma (PC). Molecular genetic testing conducted on puncture aspirates from 26 patients before surgery. The diagnosis was verified according to the morphological investigations. Mutations in BRAF V600E detected only in patients with the thyroid PC. Thus, the definition of BRAF V600E mutation may be a marker in the preoperative diagnosis of thyroid PC.

  20. Status of carcinoma cervix and high risk HPV 16 DNA in women with postmenopausal uterine bleeding (PMB

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    Veena Kashyap

    2014-09-01

    Full Text Available Postmenopausal bleeding (PMB is a discharge that occurs following the firm diagnosis of menopause, which is at least six months from the end of women’s menstrual cycle but not to be confused with infrequent or irregular periods occurring around the time of menopause. It is a common problem representing 5% of all gynecology outpatient attendances which are to eliminate endometrial cancer as the cause of bleed and PMB should be reported urgently to the gynecologist. Uterine bleeding in postmenopausal women is highly indicative clinically of malignancy originating from cervix or endometrium and Human papilloma virus (HPV is one of the causative agent for carcinoma cervix. Incidence of carcinoma cervix increases with the age in mature women, however, incidence of human papillomavirus (HPV infection reduces as menopause sets in. The presence of the virus could be used as an early indication of disease potential. Because the Pap test can only detect clinical evidence of cervical disease, molecular-based diagnostic tools are being used more frequently to detect the virus before abnormal cell growth can be observed. This study was aimed to determine the status of cervical cancer and HPV 16 DNA positivity in relation to postmenopausal bleeding.

  1. Different Effects of Homocysteine and Oxidized Low Density Lipoprotein on Methylation Status in the Promoter Region of the Estrogen Receptor α Gene

    Institute of Scientific and Technical Information of China (English)

    Yushan HUANG; Kejun PENG; Juan SU; Yuping HUANG; Yizhou XU; Shuren WANG

    2007-01-01

    We investigated the effects of homocysteine (Hcy) and oxidized low density lipoprotein (oxLDL) on DNA methylation in the promoter region of the estrogen receptor α (ERα) gene, and its potential mechanism in the pathogenesis of atherosclerosis. Cultured smooth muscle cells (SMCs) of humans were treated by Hcy and ox-LDL with different concentrations for different periods of time. The DNA methylation status was assayed by nested methylation-specific polymerase chain reaction, the lipids that accumulated in the SMCs and foam cell formations were examined with Oil red O staining. The proliferation of SMCs was assayed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. The results showed that ox-LDL in moderate concentrations (10-40 mg/L) induced de novo methylation in the promoter region of the ERα gene of SMCs. However, high concentrations (50 mg/L) of ox-LDL, resulted in demethylation of ERα. The Hcy treatment resulted in de novo methylation in the promoter region of the ERα gene with a concentration- and treating time-dependent manner, and a dose-dependent promoting effect on SMC proliferation. These data indicated that the two risk factors for atherosclerosis had the function of inducing de novo methylation in the promoter region of the ERα gene of SMCs. However, high concentrations (50mg/L) of ox-LDL induced demethylation, indicating that different risk factors of atherosclerosis with different potency might cause different aberrant methylation patterns in the promoter region of the ERα gene. The atherogenic mechanism of Hcy might involve the hypermethylation of the ERα gene, leading to the proliferation of SMCs in atherosclerotic lesions.

  2. TP53 Promoter Methylation in Primary Glioblastoma: Relationship with TP53 mRNA and Protein Expression and Mutation Status

    OpenAIRE

    JESIONEK-KUPNICKA, DOROTA; Szybka, Malgorzata; Malachowska, Beata; Fendler, Wojciech; Potemski, Piotr; Piaskowski, Sylwester; Jaskolski, Dariusz; Papierz, Wielislaw; Skowronski, Wieslaw; Och, Waldemar; Kordek, Radzislaw; ZAWLIK, IZABELA

    2014-01-01

    Reduced expression of TP53 by promoter methylation has been reported in several neoplasms. It remains unclear whether TP53 promoter methylation is associated with reduced transcriptional and protein expression in glioblastoma (GB). The aim of our work was to study the impact of TP53 methylation and mutations on TP53 mRNA level and protein expression in 42 molecularly characterized primary GB tumors. We also evaluate the impact of all molecular alterations on the overall patient survival. The ...

  3. [Correlation of size of the primary tumor and axillary node status with the p53 tumor suppressor gene in carcinoma of the breast].

    Science.gov (United States)

    Topić, Brano; Stanković, N; Savjak, D; Grbić, S

    2002-01-01

    Correlation of standard pathomorphological prognostic parameters, primary tumor size and axillary nodal status with new prognostic factor in breast carcinoma: tumor suppressor gene p53 was analyzed. The studied sample included 65 women who underwent surgery for breast carcinoma at the Surgical Clinic of Clinical Center Banja Luka, from January 1st 1997 till January 1st 1999. Statistical data analysis was performed and correlation of prognostic factors was determined. The majority of authors in this field agree that the primary tumor size and axillary nodal status are the two most important prognostic factors. These factors are the best predictors of prognosis and survival of women who had the tumor and were operated on. Tumor markers were immunohistochemically determined in the last ten years and, according to the majority of authors, are still considered the additional or relative prognostic factors in breast carcinoma. Their prognostic value and significance increase almost daily. Most frequently determined tumor markers are bcl-2, pS2, Ki-67 and p53. There was a positive, directly proportional relationship between primary tumor size and tumor suppressor gene p53, but there was no positive correlation between the axillary nodal status and tumor suppressor gene p53. Significance of determination of new tumor markers as the prognostic factors was emphasized. These markers represent a powerful tool in the early detection and prevention of breast carcinoma.

  4. Clinical studies of combined photodynamic therapy using 5-fluorouracil and methyl-aminolevulinate in patients at high risk for squamous cell carcinoma

    Science.gov (United States)

    Maytin, Edward V.; Lohser, Sara; Tellez, Alejandra; Wene, Lauren; Ishak, Rim; Anand, Sanjay

    2013-03-01

    Photodynamic therapy (PDT) using aminolevulinic acid or its methyl ester, methyl-aminolevulinate (MAL), is an increasingly recognized approach for treating squamous neoplasia of the skin. Advantages of MAL-PDT include its ability to cover broad diseased areas (field treatment), and to do multiple sessions with little-to-no risk of scarring or mutagenesis. MAL-PDT is especially valuable in certain populations at high risk for skin cancer, including Caucasian patients with extensive solar damage, and organ transplant recipients (OTR) who take immunosuppressive drugs to prevent graft rejection. The latter group has a 65-200 fold increased risk of developing squamous cell carcinoma (SCC), a major cause of mortality. Therapeutic options for those patients, other than frequent surgeries, are very limited. Topical 5-Fluorouracil (5-FU), frequently prescribed in normal patients for pre-SCC of the skin, is only minimally effective in the OTR group. MAL-PDT, however, has ~40% efficacy for pre-SCC in OTR patients. Based upon our preclinical studies in mouse tumor models, which showed that preconditioning with 5-FU can drive higher accumulation of target protoporphyins (PpIX), we proposed a rational combination regimen of 5-FU and MAL-PDT in humans. A clinical trial was designed to test the hypothesis that a combination of 5-FU followed by MAL-PDT will elevate PpIX levels and achieve better clinical outcomes in high-risk OTR patients. Primary endpoints include PpIX levels and biochemical markers (p53) measured noninvasively and in skin biopsies. Lesion clearance and recurrence (via photographs and clinical exam) are secondary endpoints. Ongoing results of this clinical trial are presented.

  5. Diet folate, DNA methylation and genetic polymorphisms of MTHFR C677T in association with the prognosis of esophageal squamous cell carcinoma

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    Shen Hongbing

    2011-03-01

    Full Text Available Abstract Background Folic acid may affect the development of human cancers. However, few studies have evaluated the consumption of diet folate in the prognosis of patients with esophageal squamous cell carcinoma (ESCC. Methods One hundred and twenty five ESCC patients underwent esophagectomy between January 2005 and March 2006 in the Yangzhong People's Hospital were recruited and followed up. The effects of diet folate, aberrant DNA methylation of selected genes and methylenetetrahydrofolate reductase (MTHFR C677T genetic polymorphisms on the prognosis of ESCC were evaluated by using Cox proportional hazard regression models. Results Our analysis showed an inverse association between diet folate intake and the risk of death after esophagectomy. The median survival time was 3.06 years for low or moderate folate consumption and over 4.59 years for high folate consumption. After adjusting for potential confounders, the hazard ratios (95% confidence interval [HRs (95% CI] were 0.72 (0.36-1.46 for moderate and 0.39 (0.20-0.78 for high folate intake, respectively (P for trend = 0.007. This preventive effect was more evident in patients carrying MTHFR 677CC genotype. No significant relation was observed between aberrant DNA methylation of P16, MGMT and hMLH1 gene, as well as MTHFR C677T genetic polymorphisms and the prognosis of ESCC. Conclusions Our research indicated that diet folate intake may have benefits on the prognosis of ESCC after esophagectomy. From a practical viewpoint, the findings of our study help to establish practical intervention and surveillance strategies for managements of ESCC patients and can finally decrease the disease burden.

  6. Maternal nutritional status, C(1) metabolism and offspring DNA methylation: a review of current evidence in human subjects.

    Science.gov (United States)

    Dominguez-Salas, Paula; Cox, Sharon E; Prentice, Andrew M; Hennig, Branwen J; Moore, Sophie E

    2012-02-01

    Evidence is growing for the long-term effects of environmental factors during early-life on later disease susceptibility. It is believed that epigenetic mechanisms (changes in gene function not mediated by DNA sequence alteration), particularly DNA methylation, play a role in these processes. This paper reviews the current state of knowledge of the involvement of C1 metabolism and methyl donors and cofactors in maternal diet-induced DNA methylation changes in utero as an epigenetic mechanism. Methyl groups for DNA methylation are mostly derived from the diet and supplied through C1 metabolism by way of choline, betaine, methionine or folate, with involvement of riboflavin and vitamins B6 and B12 as cofactors. Mouse models have shown that epigenetic features, for example DNA methylation, can be altered by periconceptional nutritional interventions such as folate supplementation, thereby changing offspring phenotype. Evidence of early nutrient-induced epigenetic change in human subjects is scant, but it is known that during pregnancy C1 metabolism has to cope with high fetal demands for folate and choline needed for neural tube closure and normal development. Retrospective studies investigating the effect of famine or season during pregnancy indicate that variation in early environmental exposure in utero leads to differences in DNA methylation of offspring. This may affect gene expression in the offspring. Further research is needed to examine the real impact of maternal nutrient availability on DNA methylation in the developing fetus.

  7. Carpel, a new Arabidopsis epi-mutant of the SUPERMAN gene: phenotypic analysis and DNA methylation status.

    Science.gov (United States)

    Rohde, A; Grunau, C; De Beck, L; Van Montagu, M; Rosenthal, A; Boerjan, W

    1999-09-01

    The carpel (car) mutation affects the morphology of reproductive organs in Arabidopsis thaliana. car flowers have an increased number of carpels, on average 2.7 +/- 0.8 instead of two in the wild type. Through allelism test with fon1-3 and analysis of the methylation state of the SUPERMAN (SUP) gene in car mutants, we show that car is an epi-mutation of SUP. The methylation pattern of car is clearly distinct from that of fon1-3, another epi-mutation of the SUP gene. Methylation was found predominantly in Cp(A/T)p(A/G) triplets and in CpG pairs. We suggest that the extensive SUP methylation in car has arisen from an abundant methylation of a single CpG site that was already present in abscisic acid-insensitive (abi3-4) mutants, from which car was segregating.

  8. TMEM16A/ANO1 is differentially expressed in HPV-negative versus HPV-positive head and neck squamous cell carcinoma through promoter methylation.

    Science.gov (United States)

    Dixit, Ronak; Kemp, Carolyn; Kulich, Scott; Seethala, Raja; Chiosea, Simion; Ling, Shizhang; Ha, Patrick K; Duvvuri, Umamaheswar

    2015-11-13

    Head and neck squamous cell carcinoma (HNSCC) has a variety of causes. Recently, the human papilloma virus (HPV) has been implicated in the rising incidence of oropharyngeal cancer and has led to variety of studies exploring the differences between HPV-positive and HPV-negative HNSCC. The calcium-activated chloride channel TMEM16A is overexpressed in a variety of cancers, including HNSCC, but whether or not it plays different roles in HPV-positive and HPV-negative HNSCC is unknown. Here, we demonstrate that TMEM16A is preferentially overexpressed in HPV-negative HNSCC and that this overexpression of TMEM16A is associated with decreased patient survival. We also show that TMEM16A expression is decreased in HPV-positive HNSCC at the DNA, RNA, and protein levels in patient samples as well as cell lines. We demonstrate that the lower levels of TMEM16A expression in HPV-positive tumors can be attributed to both a combination of copy number alteration and promoter methylation at the DNA level. Additionally, our cellular data show that HPV-negative cell lines are more dependent on TMEM16A for survival than HPV-positive cell lines. Therefore, we suspect that the down-regulation of TMEM16A in HPV-positive HNSCC makes TMEM16A a poor therapeutic target in HPV-positive HNSCC, but a potentially useful target in HPV-negative HNSCC.

  9. Sentinel lymph node biopsy in oral and oropharyngeal squamous cell carcinoma: current status and unresolved challenges

    Energy Technology Data Exchange (ETDEWEB)

    Bluemel, Christina; Herrmann, Ken [University Hospital of Wuerzburg, Department of Nuclear Medicine, Wuerzburg (Germany); Rubello, Domenico [Rovigo Hospital, Department of Nuclear Medicine-PET/CT Oncologic and Endocrine Sections, Rovigo (Italy); Colletti, Patrick M. [University of Southern California, Department of Radiology, Los Angeles, CA (United States); Bree, Remco de [UMC Utrecht Cancer Center, Department of Head and Neck Surgical Oncology, Utrecht (Netherlands)

    2015-08-15

    Because imaging with ultrasound, computed tomography, magnetic resonance imaging or positron emission tomography is unreliable for preoperative lymph node staging of early-stage oral and oropharyngeal squamous cell carcinoma (OSCC), elective neck dissection has been typically performed. The targeted sampling of sentinel lymph nodes (SLN) identified by lymphoscintigraphy and detected by gamma probe has become an effective alternative for the selection of patients for regional nodal resection. With careful consideration to technique, high SLN detection rates have been reported. Advanced techniques including intraoperative handheld gamma camera imaging and freehand single photon emission computed tomography (SPECT) are expected to increase surgical confidence in these procedures. This review gives an update on SLN biopsy in patients with OSCC including clinical standards and controversial aspects. (orig.)

  10. Expression of the p53 target Wig-1 is associated with HPV status and patient survival in cervical carcinoma.

    Directory of Open Access Journals (Sweden)

    Li-Di Xu

    Full Text Available The p53 target gene WIG-1 (ZMAT3 is located in chromosomal region 3q26, that is frequently amplified in human tumors, including cervical cancer. We have examined the status of WIG-1 and the encoded Wig-1 protein in cervical carcinoma cell lines and tumor tissue samples. Our analysis of eight cervical cancer lines (Ca Ski, ME-180, MS751, SiHa, SW756, C-4I, C-33A, and HT-3 by spectral karyotype, comparative genomic hybridization and Southern blotting revealed WIG-1 is not the primary target for chromosome 3 gains. However, WIG-1/Wig-1 were readily expressed and WIG-1 mRNA expression was higher in the two HPV-negative cervical cell lines (C33-A, HT-3 than in HPV-positive lines. We then assessed Wig-1 expression by immunohistochemistry in 38 cervical tumor samples. We found higher nuclear Wig-1 expression levels in HPV-negative compared to HPV positive cases (p = 0.002 and in adenocarcinomas as compared to squamous cell lesions (p<0.0001. Cases with moderate nuclear Wig-1 staining and positive cytoplasmic Wig-1 staining showed longer survival than patients with strong nuclear and negative cytoplasmic staining (p = 0.042. Nuclear Wig-1 expression levels were positively associated with age at diagnosis (p = 0.023 and histologic grade (p = 0.034. These results are consistent with a growth-promoting and/or anti-cell death function of nuclear Wig-1 and suggest that Wig-1 expression can serve as a prognostic marker in cervical carcinoma.

  11. Beyond biology: the impact of marital status on survival of patients with adrenocortical carcinoma

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    Zachary Klaassen

    2015-12-01

    Full Text Available Purpose: To analyze the association of marital status and survival of patients with ACC using a population-based database. Material and Methods: Patients with ACC were abstracted from the Surveillance Epidemiology and End Results (SEER database from 1988-2010 (n=1271. Variables included marital status (married vs single/divorced/widowed (SDW, gender, age, race, tumor (T and node (N classification, receipt of surgery, and SEER stage. Statistical analysis was performed using Cox proportional hazard models to generate hazard ratios and 95% confidence intervals. Results: There were 728 (57.3% females and median age was 56 years (IQR 44-66. Patients who were alive were more frequently married (65.6% vs 61.6%, p=0.008, female (61.1% vs 58.0%, p=0.001, younger (median 51 vs 57 years, p=0.0001, submitted to adrenalectomy (88.6% vs 63.8%, p<0.0001, and more favorable SEER stage (localized-64.9% vs 29.9%; regional–25.1% vs 30.1%; distant 4.8% vs 31.5%, p<0.0001 compared to patients dead of disease (DOD. On multivariable analysis, factors significantly associated with all-cause mortality were SDW status (HR 1.28, 95% CI 1.091.51, age, non-operative management, and N+ disease. Risk factors for disease-specific mortality included SDW status (HR 1.30, 95% CI 1.07-1.56, age, non-operative management, T-classification, and N+ disease. Conclusions: Marital status is significantly associated with survival in patients with ACC. Our results suggest that the decreased survival seen among SDW individuals highlights an area for further research and needed intervention to reduce disparity.

  12. HER2 status in ovarian carcinomas: a multicenter GINECO study of 320 patients.

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    Marianne Tuefferd

    Full Text Available BACKGROUND: Despite a typically good response to first-line combination chemotherapy, the prognosis for patients with advanced ovarian cancer remains poor because of acquired chemoresistance. The use of targeted therapies such as trastuzumab may potentially improve outcomes for patients with ovarian cancer. HER2 overexpression/amplification has been reported in ovarian cancer, but the exact percentage of HER2-positive tumors varies widely in the literature. In this study, HER2 gene status was evaluated in a large, multicentric series of 320 patients with advanced ovarian cancer, including 243 patients enrolled in a multicenter prospective clinical trial of paclitaxel/carboplatin-based chemotherapy. METHODOLOGY/PRINCIPAL FINDINGS: The HER2 status of primary tumors and metastases was evaluated by both immunohistochemistry (IHC and fluorescence in situ hybridization (FISH analysis of paraffin-embedded tissue on conventional slides. The prognostic impact of HER2 expression was analyzed. HER2 gene was overexpressed and amplified in 6.6% of analyzed tumors. Despite frequent intratumoral heterogeneity, no statistically significant difference was detected between primary tumors and corresponding metastases. CONCLUSIONS/SIGNIFICANCE: Our results show that the decision algorithm usually used in breast cancer (IHC as a screening test, with equivocal results confirmed by FISH is appropriate in ovarian cancer. In contrast to previous series, HER2-positive status did not influence outcome in the present study, possibly due to the fact that patients in our study received paclitaxel/carboplatin-based chemotherapy. This raises the question of whether HER2 status and paclitaxel sensitively are linked.

  13. Current Status of Studies on Targeted Therapy for Renal Cell Carcinoma

    Institute of Scientific and Technical Information of China (English)

    Shaoqi Wang; Shaoxiang Wang; Juan Wang

    2008-01-01

    Renal cell carcinoma (RCC) is regarded as one of the most refractory malignancies. A further study of the molecular mechanism of RCC formation has led to a series of successful examples for treatment of patients with advanced RCC. Over the past 20 years, a nonspecific immunotherapy, with cytokines, has been employed as the gold standard for therapy of metastatic RCC. However, with scientific development and clinical testing of new drugs, targeted molecular cancer therapy has become a focus of interest. At the same time, with a better understanding of RCC,the treatment method has converged on anti-vascular endothelial growth factor (VEGF) and related molecular-targeted pathways.A large amount of research and numerous clinical trials have demonstrated the clinical efficacy of the targeted molecular therapies in patients with metastatic RCC. For example sorafenib and sunitinib were approved, in 2005 and 2006 respectively, by the U.S. FDA for treating advanced RCC. In this report, issues such as the importance of VEGF in RCC and the studies of bevacizumab,sunitinib and sorafenib in treating metastatic RCC etc., are reviewed.

  14. Immunotherapeutic approaches in biliary tract carcinoma:Current status and emerging strategies

    Institute of Scientific and Technical Information of China (English)

    Eric; I; Marks; Nelson; S; Yee

    2015-01-01

    For biliary tract carcinoma(BTC),complete surgical resection of tumor is only feasible in a minority of patients,and the treatment options for patients with unresectable or metastatic disease are limited.Advances in cancer immunology have led to identification of tumor-infiltrating immune cells as indicators of prognosis and response to treatment in BTC.This has also facilitated development of immunotherapy that focuses on enhancing the immune system against biliary tumors.This includes peptide- and dendritic cell-based vaccines that stimulate in-vivo immune responses against tumorspecific antigens.Adoptive immunotherapy,which entails the ex-vivo expansion of tumor-infiltrating immune cells for subsequent reintroduction,and cytokinebased therapies have been developed in BTC.Clinical studies indicate that this type of therapy is generally well tolerated.Combination therapy with dendritic cell-based vaccines and adoptive immunotherapy has shown particularly good potential.Emerging strategies through discovery of novel antigen targets and by reversal of tumor-associated immunosuppression are expected to improve the efficacy of immunotherapy in BTC.Collaborative efforts by integration of targeted immunotherapeutics with molecular profiling of biliary tumor will hopefully make a positive impact on advancing towards the goal of developing precision treatment of patients with this highly lethal disease.

  15. DNA methylation status of epithelial-mesenchymal transition (EMT--related genes is associated with severe clinical phenotypes in ulcerative colitis (UC.

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    Tomomitsu Tahara

    Full Text Available BACKGROUND: Epithelial-to-mesenchymal transition (EMT is a phenomenon that allows the conversion of adherent epithelial cells to a mesenchymal cell phenotype, which enhances migratory capacity and invasiveness. Recent studies have suggested that EMT contributes to the pathogenesis of ulcerative colitis (UC. We investigated the promoter DNA methylation status of EMT-related genes in the colonic mucosa in UC. METHODS: Colonic biopsies were obtained from the rectal inflammatory mucosa of 86 UC patients and the non-inflammatory proximal colonic mucosa of 10 paired patients. Bisulfite pyrosequencing was used to quantify the methylation of 5 candidate CpG island promoters (NEUROG1, CDX1, miR-1247, CDH1, and CDH13 and LINE1. RESULTS: Using an unsupervised hierarchical clustering analysis, inflamed rectal mucosa was well separated from mucosa that appeared normal. The CDH1 and CDH13 promoters were significantly associated with patient age (p = 0.04, 0.03, respectively. A similar trend was found between those genes and the duration of disease (CDH1: p = 0.07, CDH13: p = 0.0002, mean of both: p<0.00001. Several positive associations were found between hypermethylation and severe clinical phenotypes (CDX1 and miR-1247 and a refractory phenotype: p = 0.04 and 0.006, respectively. miR-1247 and CDH1 hyper methylation and a more severe Mayo endoscopic subscore: miR-1247: p = 0.0008, CDH1: p = 0.03, mean of both: p = 0.003. When the severe clinical phenotype was defined as having any of five phenotypes (hospitalized more than twice, highest Mayo endoscopic subscore, steroid dependence, refractory, or a history of surgery miR-1247 hypermethylation was associated with the same phenotype (p = 0.008. CONCLUSIONS: Our data suggest that variability in the methylation status of EMT-related genes is associated with more severe clinical phenotypes in UC.

  16. Genetics of hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Andreas Teufel; Frank Staib; Stephan Kanzler; Arndt Weinmann; Henning Schulze-Bergkamen; Peter R Galle

    2007-01-01

    The completely assembled human genome has made it possible for modern medicine to step into an era rich in genetic information and high-throughput genomic analysis. These novel and readily available genetic resources and analytical tools may be the key to unravel the molecular basis of hepatocellular carcinoma (HCC). Moreover, since an efficient treatment for this disease is lacking, further understanding of the genetic background of HCC will be crucial in order to develop new therapies aimed at selected targets. We report on the current status and recent developments in HCC genetics. Special emphasis is given to the genetics and regulation of major signalling pathways involved in HCC such as p53, Wntsignalling, TGFβ, Ras, and Rb pathways. Furthermore, we describe the influence of chromosomal aberrations as well as of DNA methylation. Finally, we report on the rapidly developing field of genomic expression profiling in HCC, mainly by microarray analysis.

  17. HLA class I and II expression in oropharyngeal squamous cell carcinoma in relation to tumor HPV status and clinical outcome.

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    Anders Näsman

    Full Text Available HPV-DNA positive (HPVDNA+ oropharyngeal squamous cell carcinoma (OSCC has better clinical outcome than HPV-DNA negative (HPVDNA- OSCC. Current treatment may be unnecessarily extensive for most HPV+ OSCC, but before de-escalation, additional markers are needed together with HPV status to better predict treatment response. Here the influence of HLA class I/HLA class II expression was explored. Pre-treatment biopsies, from 439/484 OSCC patients diagnosed 2000-2009 and treated curatively, were analyzed for HLA I and II expression, p16(INK4a and HPV DNA. Absent/weak as compared to high HLA class I intensity correlated to a very favorable disease-free survival (DFS, disease-specific survival (DSS and overall survival (OS in HPVDNA+ OSCC, both in univariate and multivariate analysis, while HLA class II had no impact. Notably, HPVDNA+ OSCC with absent/weak HLA class I responded equally well when treated with induction-chemo-radiotherapy (CRT or radiotherapy (RT alone. In patients with HPVDNA- OSCC, high HLA class I/class II expression correlated in general to a better clinical outcome. p16(INK4a overexpression correlated to a better clinical outcome in HPVDNA+ OSCC. Absence of HLA class I intensity in HPVDNA+ OSCC suggests a very high survival independent of treatment and could possibly be used clinically to select patients for randomized trials de-escalating therapy.

  18. Improved survival of patients with hepatocellular carcinoma and disparities by age, race, and socioeconomic status by decade, 1983–2012

    Science.gov (United States)

    Li, Jie; Hong, Guobin; Li, Dan; Mallampati, Saradhi; Zhou, Xiuling; Zhou, Cuiling; Zhang, Hongyu; Cheng, Zhibin; Shan, Hong; Ma, Haiqing

    2016-01-01

    Hepatocellular carcinoma (HCC), accounting for the majority of liver cancer, is a highly aggressive malignancy with poor prognosis and therefore adds up the financial burden. Incidence data of HCC in three decades during 1983-2012 were extracted from the Surveillance, Epidemiology, and End Results (SEER) database with incidence rates of 1.9, 3.1 and 4.9 per 100,000 respectively. In addition, to evaluate the survival changes in the same period, a total of 63,640 HCC cancer cases were accessed from SEER database. The six-month relative survival rates improved each decade from 31.0% to 42.9% to 57.2% and the higher increase can be seen in the last two decades. More importantly, the disparities of survival among different racial groups and socioeconomic status (SES) were confirmed by the inferiority of survival in Black race and high-poverty group respectively. This research analyzed the incidence and survival data of HCC in the past three decades and may help predict the future trends of incidence and survival. Furthermore, this study may help better design healthcare policies and clinical management programs to balance the disparities of survival between SES groups, races, ages and sexes confirmed in this study and thereby improve the clinical management of HCC. PMID:27486977

  19. Anticancer Activity of Marine Sponge Hyrtios sp. Extract in Human Colorectal Carcinoma RKO Cells with Different p53 Status

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    Hyun Kyung Lim

    2014-01-01

    Full Text Available Drug development using marine bioresources is limited even though the ocean occupies about 70% of the earth and contains a large number of biological materials. From the screening test of the marine sponge extracts, we found Hyrtios sp. sponge collected from Chuuk island, Micronesia. In this study, the Hyrtios sp. extract was examined for anticancer activity against human colorectal carcinoma RKO cells that are wildtype for p53 and RKO-E6 that are p53 defective. The Hyrtios sp. extract dose-dependently inhibited viability in both cell lines. Multinucleation as an indication of mitotic catastrophe was also observed. Cytotoxicity tests gave significantly different results for RKO and RKO-E6 cells after 48 h exposure to Hyrtios sp. extract. In RKO cells treated with Hyrtios sp. extract, cell death occurred by induction of p53 and p21 proteins. In p53-defective RKO-E6 cells, Hyrtios sp. extract decreased expression of JNK protein and increased p21 protein. These results indicate that Hyrtios sp. extract induced apoptosis via different pathways depending on p53 status and could be a good natural product for developing new anticancer drugs.

  20. Evaluation of antitumor activity and antioxidant status of Alternanthera brasiliana against Ehrlich ascites carcinoma in Swiss albino mice

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    Pavan Kumar Samudrala

    2015-01-01

    Full Text Available Objective : The main objective of the present study was to explore the antitumor activity of the ethyl acetate extract of the Alternanthera brasiliana (EAAB and its antioxidant status against Ehrlich ascites carcinoma (EAC in Swiss albino mice. Materials and Methods: Based on the preliminary in vitro cytotoxicity studies, EAAB was selected for anti-tumor and antioxidant effects. Anticancer activity of EAAB was evaluated against EAC in Swiss albino mice at the doses of 200 and 400 mg/kg. EAAB was administered for 14 consecutive days after induction of cancer. After 24 h of the last dose and 18 h of fasting, half of the mice were sacrificed and rest were kept alive for assessing any increase in life span. The antitumor effect of EAAB was assessed by evaluating tumor volume, viable and nonviable tumor cell count, tumor weight, hematological and biochemical parameters of EAC bearing host. Furthermore, the antioxidant and histopathological parameters were evaluated. Results: EAAB treatment has shown significant decrease in tumor volume, viable cell count, tumor weight and elevated the life span of EAC tumor bearing mice in a dose dependent manner. In hematological profile count of RBC, hemoglobin, and WBC were found reverted to normal. EAAB also significantly decreased the levels of lipid peroxidation and significantly increased the levels of GSH, SOD and Catalase. Conclusion: From the above results it may be concluded that EAAB has potent dose dependent antitumor activity and is comparable to that of 5-flourouracil.

  1. Status quo of chronic liver diseases, including hepatocellular carcinoma, in Mongolia.

    Science.gov (United States)

    Jazag, Amarsanaa; Puntsagdulam, Natsagnyam; Chinburen, Jigjidsuren

    2012-06-01

    Because Mongolia has much higher liver disease burden than any other regions of the world, it is necessary to provide information on real-time situation of chronic liver disease in Mongolia. In this article, we reviewed studies performed in Mongolia from 2000 to 2011 on seroprevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) among healthy individuals and patients with chronic liver diseases, and on the practice patterns for the management of liver cirrhosis and hepatocellular carcinoma (HCC). According to previous reports, the seroprevalence of HBV and HCV in general population in Mongolia is very high (11.8% and 15% for HBV and HCV, respectively). Liver cirrhosis is also highly prevalent, and mortality from liver cirrhosis remained high for the past decade (about 30 deaths per 100,000 populations per year). Among patients with cirrhosis, 40% and 39% are positive for HBsAg and anti-HCV, respectively, and 20% are positive for both. The seroprevalence is similar for HCC and more than 90% of HCC patients are positive for either HBV or HCV. The incidence of HCC in Mongolia is currently among the highest in the world. The mortality from HCC is also very high (52.2 deaths per 100,000 persons per year in 2010). Partly due to the lack of established surveillance systems, most cases of HCC are diagnosed at an advanced stage. The mortality from liver cirrhosis and HCC in Mongolia may be reduced by implementation of antiviral therapy program and control of alcohol consumption.

  2. Immune-checkpoint status in penile squamous cell carcinoma: a North American cohort.

    Science.gov (United States)

    Cocks, Margaret; Taheri, Diana; Ball, Mark W; Bezerra, Stephania M; Del Carmen Rodriguez, Maria; Ricardo, Bernardo F P; Bivalacqua, Trinity J; Sharma, Rajni B; Meeker, Alan; Chaux, Alcides; Burnett, Arthur L; Netto, George J

    2017-01-01

    Penile squamous cell carcinoma (SCC) is primarily treated by surgical resection. Locally advanced and metastatic diseases require a multidisciplinary treatment approach. However, mortality and morbidity remain high, and novel molecular and immunotherapeutic targets are actively being sought. We investigated the expression of immune-checkpoint markers in penile cancers. Fifty-three invasive penile SCCs diagnosed between 1985 and 2013 were retrieved from our surgical pathology archives. Representative formalin-fixed, paraffin-embedded archival blocks were used for the construction of 2 high-density tissue microarrays. Tissue microarrays were stained with immunohistochemistry for PD-L1, FOXP3, CD8, and Ki-67. PD-L1 was investigated using rabbit monoclonal anti-PD-L1 antibody (Cell Signaling, Boston, MA; E1L3N, 1:100). Overall, 21 (40%) of 53 penile SCCs had positive PD-L1 expression. PD-L1 was expressed by a significant proportion of advanced penile SCC. Forty-four percent (15/34) of stage pT2 or more SCC and 38% (6/16) of tumors with lymph node metastasis were positive for PD-L1. PD-L1 expression did not correlate with patient age, tumor location, histologic subtype, tumor stage, anatomic depth of invasion, or tumor grade. FOXP3 expression in tumoral immune cells was found in 26 (49%) of 53 cases. FOXP3 expression in stromal immune cells correlated with tumor thickness (P = .0086). The ratio of CD8/FOXP3 was greater than 1 in 62% of cases in tumor-infiltrating immune cells and 34% of cases in stromal immune cells. Our current study is the largest to assess expression of PD-L1 in a clinically well-annotated North American cohort of penile SCC. Our findings support a rationale for targeting immune-checkpoint inhibitor pathways in advanced penile SCC.

  3. 食管鳞癌细胞中五聚素3基因的表达及甲基化%Methylation and expression of pentraxin3 gene in human esophageal squamous cell carcinoma cell lines

    Institute of Scientific and Technical Information of China (English)

    王俊雄; 姜春燕; 李敏; 朱圣韬; 张澍田

    2012-01-01

    目的 研究食管鳞状细胞癌(ESCC)中五聚素3(PTX3)基因的表达以及基因启动子区甲基化状态对其表达的影响.方法 采用RT-PCR、甲基化特异性PCR(MSP)、亚硫酸氢盐测序等方法 对ESCC细胞进行检测PTX3基因mRNA的表达、基因启动子区甲基化状态.应用甲基转移酶抑制剂5-氮-2′-脱氧胞苷(5-Aza-DC)去甲基化处理对该基因表达的影响.结论 RT-PCR结果 显示PTX3 mRNA在细胞株TE-11、EC109、EC9706、KYSE30、KYSE510均不表达,在KYSE410,Het-1A细胞株中有PTX3 mRNA表达.MSP结果 显示TE-11、EC109、EC9706、KYSE30、KYSE510存在PTX3基因甲基化,而KYSE410,Het-1A细胞株中不存在PTX3基因甲基化.甲基化测序结果 进一步证实了MSP结果 结果 ,28个CpG位点中,TE-11、EC109、EC9706、KYSE30、KYSE510甲基化程度分别为64.3%,81.0%,94.5%,78.5%及81.0%,而KYSE410,Het-1A不存在甲基化.ESCC细胞株PTX3基因启动子处于异常的高甲基化状态,经5-Aza-DC处理后,高甲基化的PTX3基因启动子去甲基化,处理前PTX3基因启动子高甲基化的细胞株其mRNA均不表达,去甲基化处理后能重新激活该基因表达.结论 PTX3基因启动子高甲基化是PTX3表达失活的主要原因之一,PTX3基因启动子甲基化检测可作为ESCC的潜在肿瘤标志物之一.%Objective To investigate the methylation and expression of PTX3 gene in esophageal squamous cell carcinoma ( ESCC) cell lines. Methods Six ESCC cell lines ( EC109 , EC9706 , KYSE30 , KYSE410 , KYSE 510, TE-11)and one esophageal epithelial cell Het-1A were selected. RT-PCR and MSP were used to detect the mRNA expression and methylation status of PTX3 gene. The esophageal cancer cell lines were treated with 5-Aza-CdR, and the changes of PTX3 mRNA expression before and after 5-Aza-CdR treatment were tested. Results PTX3 transcript was silenced in 5ESCC cell lines ( TE11 , KYSE30, KYSE510 EC109 and EC9706 ) , but not in KYSE410 and immortalized normal esophageal

  4. Effects of N-methyl pyrrolidone on the uptake of hypericin in human bladder carcinoma and co-staining with DAPI investigated by confocal microscopy.

    Science.gov (United States)

    Saw, Constance Lay Lay; Olivo, Malini; Wohland, Thorsten; Fu, Chit Yaw; Kho, Kiang Wei; Soo, Khee Chee; Sia Heng, Paul Wan

    2007-10-01

    Photodynamic diagnosis (PDD) using hypericin (HY), a natural photosensitizer, detects bladder cancer significantly better than white light endoscopy. However, the lipophilicity of HY complicates its administration for clinical applications. Currently, pharmaceutical preparations for HY without plasma protein are being developed. Formulations containing a biocompatible solvent, N-methyl pyrrolidone (NMP) have been shown to enhance the photodynamic therapeutic effects of HY. It was recently reported that, NMP formulations of HY were able to produce significantly higher contrast for fluorescence detection of tumors than albumin-containing HY formulations. This present work hypothesizes that NMP acts both as a solvent and penetration enhancer to improve the delivery of HY into cells by increasing the permeability of cell membranes. This paper reports the use of 3-D confocal microscopy to monitor real-time uptake of HY in human carcinoma. 3-D confocal microscopy was used to investigate the possibility of nuclear localization of HY in MGH cells. The fluorescence of HY was confirmed to be emitted from HY containing cells using spectrometry. The localization of a DNA fluorescent probe 4', 6-diamidino-2-phenylindole dihydrochloride (DAPI) was used to confirm the possibility of colocalization of DAPI and HY. The colocalization analysis in the present study suggests that it was very unlikely that HY colocalized in the nucleus that was stained by DAPI. Fluorescein leakage tests showed that 1% NMP changes the permeability of cell membranes, and enhanced the delivery of HY into cells resulting in lower cell survival ratios. Thus, NMP was able to enhance the photodynamic therapeutic effects of HY on cancer cells.

  5. O 6 -methylguanine DNA methyltransferase gene promoter methylation in high-grade gliomas: A review of current status

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    Vaishali Suri

    2011-01-01

    Full Text Available Assessment of promoter methylation of the O 6 -methylguanine DNA methyltransferase (MGMT gene has recently gained importance in molecular profiling of high-grade gliomas. It has emerged not only as an important prognostic marker but also as a predictive marker for response to temozolomide in patients with newly diagnosed glioblastoma. Further, recent studies indicate that MGMT promoter methylation has strong prognostic relevance even in anaplastic (grade III gliomas, irrespective of therapy (chemotherapy or radiotherapy. This article provides an overview of its use as a predictive and prognostic biomarker, as well as the methods employed for its assessment and use in therapeutic decision making.

  6. [Current status and progress in studies on the association between obesity and DNA methylation among children and adolescents].

    Science.gov (United States)

    Gao, Y; Gao, W J; Cao, W H

    2016-08-10

    DNA methylation is one of the most commonly recognized epigenetic phenomenon, which explains how genes, environmental factors and gene-environment interaction would cause obesity, integretedly. Studies on early life obesity-related epigenetic reveal important effects that related to the programs on prevention and control of obesity. However, only few similar studies have been conducted in China. This paper summarizes the basic principles, characteristics of DNA methylation and the major results of children and adolescents obesity-related research areas, in order to provide evidence for further studies.

  7. Molecular characterization, expression and methylation status analysis of BMP4 gene in skin tissue of Liaoning cashmere goat during hair follicle cycle.

    Science.gov (United States)

    Bai, Wen L; Dang, Yun L; Wang, Jiao J; Yin, Rong H; Wang, Ze Y; Zhu, Yu B; Cong, Yu Y; Xue, Hui L; Deng, Liang; Guo, Dan; Wang, Shi Q; Yang, Shu H

    2016-08-01

    Bone morphogenetic protein 4 (BMP4) is a member of the bone morphogenetic protein family (BMPs). It is involved in the development and cycle of hair follicle, as well as, is thought to be a potential candidate gene for cashmere traits in goats. In the present study, we isolated and characterized a full-length open reading frame (ORF) of BMP4 cDNA from the skin tissue of Liaoning cashmere goat, and investigated the transcriptional pattern and methylation status of BMP4 gene in skin tissue of this breed during different stages of hair follicle cycle. The sequence analysis indicated that the isolated cDNA was 1264-bp in length containing a complete ORF of 1230-bp. It encoded a precursor peptide of 409 amino acids with a signal peptide of 19 amino acids. The structural analysis indicated that goat BMP4 contains typical TGF-β propeptide and TGF-β domains. In skin tissue, BMP4 is generally transcribed in an ascendant pattern from anagen to telogen. The methylation level of 5' flanking regulatory region of BMP4 gene might be involved in its mRNA expression in skin tissue: a higher BMP4 methylation level in skin coincides with a lower expression of BMP4 mRNA. These results from the present work provided a foundation for further insight into the functional and regulatory characteristics of BMP4 in the development and cycle of hair follicle in Liaoning Cashmere goat.

  8. Transarterial ablation of hepatocellular carcinoma. Status and developments; Transvaskulaere Ablation des hepatozellulaeren Karzinoms. Ist Chemotherapie alles

    Energy Technology Data Exchange (ETDEWEB)

    Radeleff, B.A.; Stampfl, U.; Sommer, C.M.; Bellemann, N.; Kauczor, H.U. [Universitaetsklinikum Heidelberg, Abt. Diagnostische und Interventionelle Radiologie, Radiologische Klinik, Heidelberg (Germany); Hoffmann, K. [Universitaetsklinikum Heidelberg, Abt. fuer Allgemein-, Viszeral- und Transplantationschirurgie, Chirurgische Klinik, Heidelberg (Germany); Ganten, T.; Ehehalt, R. [Universitaetsklinikum Heidelberg, Medizinische Klinik IV, Gastroenterologie, Infektionskrankheiten, Vergiftungen, Heidelberg (Germany)

    2012-01-15

    Hepatocellular carcinoma (HCC) is the fifth most common cancer worldwide and represents the main cause of death among European patients with liver cirrhosis. Only 30-40% of patients diagnosed with HCC are candidates for curative treatment options (e.g. surgical resection, liver transplantation or ablation). The remaining majority of patients must undergo local regional and palliative therapies. Transvascular ablation of HCC takes advantage of the fact that the hypervascularized HCC receives most of its blood supply from the hepatic artery. In this context transvascular ablation describes different therapy regimens which can be assigned to four groups: cTACE (conventional transarterial chemoembolization), bland embolization (transarterial embolization TAE), DEB-TACE (TACE with drug-eluting beads, DEB) and SIRT (selective internal radiation therapy, radioembolization). Conventional TACE is the most common type of transvascular ablation and represents a combination of intra-arterial chemotherapy and embolization with occlusion of the arterial blood supply. However, there is no standardized regimen with respect to the chemotherapeutic drug, the embolic agent, the usage of lipiodol and the interval between the TACE procedures. Even the exact course of a cTACE procedure (order of chemotherapy or embolization) is not standardized. It remains unclear whether or not intra-arterial chemotherapy is definitely required as bland embolization using very small, tightly calibrated spherical particles (without intra-arterial administration of a chemotherapeutic drug) shows tumor necrosis comparable to cTACE. For DEB-TACE microparticles loaded with a chemotherapeutic drug combine the advantages of cTACE and bland embolization. Thereby, a continuing chemotherapeutic effect within the tumor might cause a further increase in intratumoral cytotoxicity and at the same time a decrease in systemic toxicity. (orig.) [German] Das hepatozellulaere Karzinom (HCC) ist weltweit betrachtet das

  9. Amyloid-β alters the DNA methylation status of cell-fate genes in an Alzheimer's disease model.

    Science.gov (United States)

    Taher, Noor; McKenzie, Courtney; Garrett, Rebecca; Baker, Matthew; Fox, Nena; Isaacs, Gary D

    2014-01-01

    Alzheimer's disease (AD) is characterized by neurofibrillary tangles and extracellular amyloid-β plaques (Aβ). Despite ongoing research, some ambiguity remains surrounding the role of Aβ in the pathogenesis of this neurodegenerative disease. While several studies have focused on the mutations associated with AD, our understanding of the epigenetic contributions to the disease remains less clear. To that end, we determined the changes in DNA methylation in differentiated human neurons with and without Aβ treatment. DNA was isolated from neurons treated with Aβ or vehicle, and the two samples were digested with either a methylation-sensitive (HpaII) or a methylation-insensitive (MspI) restriction endonuclease. The fragments were amplified and co-hybridized to a commercial promoter microarray. Data analysis revealed a subset of genomic loci that shows a significant change in DNA methylation following Aβ treatment in comparison to the control group. After mapping these loci to nearby genes, we discovered high enrichment for cell-fate genes that control apoptosis and neuronal differentiation. Finally, we incorporated three of those genes in a possible model suggesting the means by which Aβ contributes to the brain shrinkage and memory loss seen in AD.

  10. Molecular characterization of HOXC8 gene and methylation status analysis of its exon 1 associated with the length of cashmere fiber in Liaoning cashmere goat.

    Science.gov (United States)

    Bai, Wen L; Wang, Jiao J; Yin, Rong H; Dang, Yun L; Wang, Ze Y; Zhu, Yu B; Cong, Yu Y; Deng, Liang; Guo, Dan; Wang, Shi Q; Yang, Shu H; Xue, Hui L

    2017-02-01

    Homeobox protein Hox-C8 (HOXC8) is a member of Hox family. It is expressed in the dermal papilla of the skin and is thought to be associated with the hair inductive capacity of dermal papilla cells. In the present study, we isolated and characterized a full-length open reading frame of HOXC8 cDNA from the skin tissue of Liaoning cashmere goat, as well as, established a phylogenetic relationship of goat HOXC8 with that of other species. Also, we investigated the effect of methylation status of HOXC8 exon 1 at anagen secondary hair follicle on the cashmere fiber traits in Liaoning cashmere goat. The sequence analysis indicated that the obtained cDNA was 1134-bp in length containing a complete ORF of 729-bp. It encoded a peptide of 242 amino acid residues in length. The structural analysis indicated that goat HOXC8 contained a typical homeobox domain. The phylogenetic analysis revealed that Capra hircus HOXC8 had a closer genetic relationship with that of Ovis aries, followed by Bos Taurus and Bubalus bubalis. The methylation analysis suggested that the methylation degree of HOXC8 exon 1 in anagen secondary hair follicle might be involved in regulating the growth of cashmere fiber in Liaoning cashmere goat. Our results provide new evidence for understanding the molecular structural and evolutionary characteristics of HOXC8 in Liaoning cashmere goat, as well as, for further insight into the role of methylation degree of HOXC8 exon 1 regulates the growth of cashmere fiber in goat.

  11. Present status and future of high-precision image guided adaptive brachytherapy for cervix carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Poetter, Richard; Kirisits, Christian; Fidarova, Elena F.; Dimopoulos, Johannes C. A.; Berger, Daniel; Tanderup, Kari; Lindegaard, Jacob C. (Dept. of Radiotherapy Medical Univ. of Vienna, General Hospital Vienna, Wien (Austria))

    2008-08-15

    Introduction. Image guided adaptive brachytherapy (IGABT) for cervical cancer, using mainly MRI, is an evolving method, increasingly replacing the 2D approach based on conventional radiography. During the complex 4D chain of this procedure image-assistance is provided for disease assessment, provisional treatment planning ('pre-planning'), applicator placement and reconstruction, as well as for contouring, definitive treatment planning and quality control of dose delivery. With IGABT changes of topography adjacent to the applicator, caused by tumour regression, oedema, organ changes and dilation are identified. Thus, the CTV for IGABT is primarily based on the tumour volume at the time of BT and takes into account both time and spatial domains. IGABT requires systematic concepts for target, OAR, biological modelling, DVH analysis, and dose-volume-adaptation. Methods and Results. This report focuses on the advantages and uncertainties, dose-effect relations and clinical results of the IGABT procedure addressing the current status and future perspectives. Uncertainties during the 4D chain of IGABT are mainly related to target contouring, applicator reconstruction, as well as to inter-fraction, intra-fraction and inter-application variability, as caused by tumour response and organ changes. Different from EBRT where set-up uncertainties are compensated by adding a margin to the CTV, no margins to the lateral and anterior-posterior directions can be used for IGABT. Discussion. By 3D treatment planning for IGABT significant improvement of the DVH parameters is achieved compared to 2D library plans. In small tumours the benefit is primarily obtained by a decrease of dose to nearby OAR while in large tumours the use of supplementary interstitial techniques and optimization may double the target volume that can be treated at a therapeutic dose level. The clinical impact of IGABT could recently be demonstrated by the establishment of some correlations between

  12. Overexpression of the mitochondrial methyltransferase TFB1M in the mouse does not impact mitoribosomal methylation status or hearing

    DEFF Research Database (Denmark)

    Lee, Seungmin; Rose, Simon; Metodiev, Metodi D;

    2015-01-01

    Mitochondrial dysfunction is a well-established cause of sensorineural deafness, but the pathophysiological events are poorly understood. Non-syndromic deafness and predisposition to aminoglycoside-induced deafness can be caused by specific mutations in the 12S rRNA gene of mtDNA and are thus...... by 'hypermethylation' of two conserved adenosines of 12S rRNA in the mitoribosome is of key pathophysiological importance in sensorineural deafness. In support for this concept, it was reported that overexpression of the essential mitochondrial methyltransferase TFB1M in the mouse was sufficient to induce...... mitoribosomal hypermethylation and deafness. At variance with this model, we show here that 12S rRNA is near fully methylated in vivo in the mouse and thus cannot be further methylated to any significant extent. Furthermore, bacterial artificial chromosome transgenic mice overexpressing TFB1M have no increase...

  13. Ultrasound-guided diffuse optical tomography (DOT) of invasive breast carcinoma: Does tumour total haemoglobin concentration contribute to the prediction of axillary lymph node status?

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, Qingli, E-mail: qinglizhu@gmail.com [Department of Diagnostic Ultrasound, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Shuaifuyuan 1, Dongcheng District, Beijing 100730 (China); Xiao, Mengsu, E-mail: xiaomengsu_2000@sina.com [Department of Diagnostic Ultrasound, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Shuaifuyuan 1, Dongcheng District, Beijing 100730 (China); You, Shanshan, E-mail: shanshan_0531@sina.com [Department of Diagnostic Ultrasound, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Shuaifuyuan 1, Dongcheng District, Beijing 100730 (China); Zhang, Jing, E-mail: zhang.jing1029@163.com [Department of Diagnostic Ultrasound, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Shuaifuyuan 1, Dongcheng District, Beijing 100730 (China); Jiang, Yuxin, E-mail: yuxinjiangxh@yahoo.com.cn [Department of Diagnostic Ultrasound, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Shuaifuyuan 1, Dongcheng District, Beijing 100730 (China); Lai, Xingjian, E-mail: lxjpumch@yahoo.com.cn [Department of Diagnostic Ultrasound, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Shuaifuyuan 1, Dongcheng District, Beijing 100730 (China); Dai, Qing, E-mail: qingdai_2000@yahoo.com [Department of Diagnostic Ultrasound, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Shuaifuyuan 1, Dongcheng District, Beijing 100730 (China)

    2012-11-15

    Objectives: To prospectively study the ultrasound-guided near-infrared diffuse optical tomography (DOT) findings of the total haemoglobin concentration (THC) detected in invasive breast carcinomas and its contribution to the prediction of axillary lymph node (LN) status. Methods: A total of 195 invasive breast carcinomas were prospectively studied with DOT before surgery. Lumpectomy or mastectomy with full axillary nodal dissection was performed. Tumour size and THC level were correlated with LN status by a logistic regression analysis. Results: One hundred twenty-four patients (63.59%) was LN(-) and 71 (36.41%) was LN(+). The average THC was significantly higher in the LN(+) group than in the LN(-) group (252.94 {+-} 69.19 {mu}mol/L versus 203.86 {+-} 83.13 {mu}mol/L, P = 0.01). A multivariate analysis showed an independent relationship between the probability of axillary metastasis, elevated THC level (P = 0.01), and tumour size (P = 0.001). The odds ratio with THC {>=} 140 {mu}mol/L was 13.651 (1.781-104.560), whereas that of tumour size with a 1 cm increment was only 1.777 (1.283-2.246). Conclusions: The THC level and the tumour size are independent and preoperative predictors of axillary nodal status; these variables may improve the diagnosis of patients with lymph node metastasis.

  14. The time-dependent accumulation of protoporphyrin IX fluorescence in nodular basal cell carcinoma following application of methyl aminolevulinate with an oxygen pressure injection device.

    Science.gov (United States)

    Blake, E; Campbell, S; Allen, J; Mathew, J; Helliwell, P; Curnow, A

    2012-12-01

    Topical protoporphyrin (PpIX)-induced photodynamic therapy (PDT) relies on the penetration of the prodrug into the skin lesion and subsequent accumulation of the photosensitizer. Methyl aminolevulinate (MAL)-PDT is an established treatment for thinner and superficial non-melanoma skin cancers (NMSCs) but for the treatment of the thicker nodular basal cell carcinoma (nBCC) enhanced penetration of the prodrug is required. This study employed a new higher pressure, oxygen pressure injection (OPI) device, at the time of Metvix® application with a view to enhancing the penetration of MAL into the tumors. Each patient had Metvix® applied to a single nBCC followed by application of a higher pressure OPI device. Following different time intervals (0, 30, 60, 120 or 180 min) the tumors were excised. The maximum depth and area of MAL penetration achieved in each lesion was measured using PpIX fluorescence microscopy. As expected, an increase in the depth of MAL-induced PpIX accumulation and area of tumor sensitized was observed over time; when the Metvix® cream was applied for 0, 30, 60, 120 and 180 min the median depth of PpIX fluorescence was 0%, 21%, 26.5%, 75.5% and 90%, respectively and the median area of tumor sensitized was 0%, 4%, 6%, 19% and 60%, respectively. As the investigation presented here did not include a control arm, the relative depths of fluorescence observed in this study were statistically compared (using the non-parametric Mann Whitney U test) with the results of our previous study where patients had Metvix® cream applied either with or without the standard pressure OPI device. When the higher pressure OPI device was employed compared to without OPI this increase was observed to be greater following 30, 120, and 180 min although overall not significantly (p=0.835). In addition, no significant difference between the higher pressure OPI device employed here and the previously investigated standard pressure OPI device was observed (p=0.403). However

  15. The Predictive but Not Prognostic Value of MGMT Promoter Methylation Status in Elderly Glioblastoma Patients: A Meta-Analysis

    OpenAIRE

    Yin, An-an; Zhang, Lu-hua; Cheng, Jin-xiang; Dong, Yu; Liu, Bo-Lin; Han, Ning; Zhang, Xiang

    2014-01-01

    Background The clinical implication of O6-methylguanine-DNA methyltransferase (MGMT) promoter status is ill-defined in elderly glioblastoma patients. Here we report a meta-analysis to seek valid evidence for its clinical relevance in this subpopulation. Methods Literature were searched and reviewed in a systematic manner using the PubMed, EMBASE and Cochrane databases. Studies investigating the association between MGMT promoter status and survival data of elderly patients (≥65 years) were eli...

  16. Feasibility of gemcitabine and oxaliplatin in patients with advanced biliary tract carcinoma and a performance status of 2.

    Science.gov (United States)

    Mir, Olivier; Coriat, Romain; Dhooge, Marion; Perkins, Géraldine; Boudou-Rouquette, Pascaline; Brezault, Catherine; Ropert, Stanislas; Durand, Jean-Philippe; Chaussade, Stanislas; Goldwasser, François

    2012-08-01

    The use of gemcitabine and oxaliplatin is well documented in selected patients with advanced biliary tract carcinoma (BTC), but little is known on the feasibility of systemic treatments in patients with a performance status (PS) of 2. We retrospectively examined the medical records of consecutive BTC patients with a PS of 2 receiving gemcitabine 1000 mg/m(2) plus oxaliplatin 100 mg/m(2) every 2 weeks from January 2003 to December 2011 in our institution. Body composition was analysed by computed tomography scan to detect sarcopenia. The primary evaluation criterion was safety. The secondary evaluation criteria were the response rate, progression-free survival (PFS) and overall survival (OS). Twenty-eight patients (median age: 63 years, range 41-83) received a total of 175 cycles (median per patient: 6, range 2-12). Ten patients (35.7%) had sarcopenia on the pretreatment computed tomography scan. The most frequent toxicities were thrombocytopenia (grades 2-4: n=4, 14.3%), peripheral neuropathy (grades 2-3: n=9, 32.1%) and cholangitis (n=4, 14.3%). The best response was a partial response in 10.7% of patients [95% confidence interval (CI): 0-22.2] and stable disease in 42.9% of patients. The median PFS and OS were 4.6 (95% CI: 2.5-6.3) and 7.5 (95% CI: 5.2-9.5) months, respectively. The median PFS and OS were significantly longer in patients without sarcopenia: 7.0 months (95% CI: 4.4-8.0) vs. 2.2 months (95% CI: 2.0-2.5), P less than 0.01, and 10.4 months (95% CI: 7.5-11.6) vs. 4.9 months (95% CI: 3.7-5.2), P less than 0.01, respectively. In our experience, gemcitabine-oxaliplatin was feasible and induced effective palliation in PS2 patients with advanced BTC. Further studies are warranted to confirm these findings.

  17. The different radiation response and radiation-induced bystander effects in colorectal carcinoma cells differing in p53 status

    Energy Technology Data Exchange (ETDEWEB)

    Widel, Maria, E-mail: maria.widel@polsl.pl [Biosystems Group, Institute of Automatic Control, Silesian University of Technology, 16 Akademicka Street, 44-100 Gliwice (Poland); Lalik, Anna; Krzywon, Aleksandra [Biosystems Group, Institute of Automatic Control, Silesian University of Technology, 16 Akademicka Street, 44-100 Gliwice (Poland); Poleszczuk, Jan [College of Inter-faculty Individual Studies in Mathematics and Natural Sciences, University of Warsaw, 93 Zwirki i Wigury Street, 02-089 Warsaw (Poland); Department of Integrated Mathematical Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, Florida (United States); Fujarewicz, Krzysztof; Rzeszowska-Wolny, Joanna [Biosystems Group, Institute of Automatic Control, Silesian University of Technology, 16 Akademicka Street, 44-100 Gliwice (Poland)

    2015-08-15

    Highlights: • We tested radiation response and bystander effect on HCT116p53+/+ and p53−/− cells. • The p53+/+ cells developed premature senescence in exposed and bystander neighbors. • Directly exposed and bystander p53−/− cells died profoundly through apoptosis. • Interleukins 6 and 8 were differently generated by both cell lines. • NFκB path was activated mainly in p53+/+ hit cells, in p53 −/− in bystanders only. - Abstract: Radiation-induced bystander effect, appearing as different biological changes in cells that are not directly exposed to ionizing radiation but are under the influence of molecular signals secreted by irradiated neighbors, have recently attracted considerable interest due to their possible implication for radiotherapy. However, various cells present diverse radiosensitivity and bystander responses that depend, inter alia, on genetic status including TP53, the gene controlling the cell cycle, DNA repair and apoptosis. Here we compared the ionizing radiation and bystander responses of human colorectal carcinoma HCT116 cells with wild type or knockout TP53 using a transwell co-culture system. The viability of exposed to X-rays (0–8 Gy) and bystander cells of both lines showed a roughly comparable decline with increasing dose. The frequency of micronuclei was also comparable at lower doses but at higher increased considerably, especially in bystander TP53-/- cells. Moreover, the TP53-/- cells showed a significantly elevated frequency of apoptosis, while TP53+/+ counterparts expressed high level of senescence. The cross-matched experiments where irradiated cells of one line were co-cultured with non-irradiated cells of opposite line show that both cell lines were also able to induce bystander effects in their counterparts, however different endpoints revealed with different strength. Potential mediators of bystander effects, IL-6 and IL-8, were also generated differently in both lines. The knockout cells secreted IL-6 at

  18. Prognostic value of MGMT methylation in colorectal cancer: a meta-analysis and literature review.

    Science.gov (United States)

    Li, Yanliang; Lyu, Zhongchuan; Zhao, Lixin; Cheng, Hong; Zhu, Dongyuan; Gao, Yongsheng; Shang, Xiuwan; Shi, Huaijie

    2015-03-01

    The development of colorectal cancer (CRC) spans about 5-10 years, making early detection and prevention beneficial to the survival of CRC patients. To address inconsistencies in evidence regarding O(6)-methylguanine-DNA-methyltransferase (MGMT) methylation as a potential prognostic factor in CRC, we conducted a meta-analysis to evaluate MGMT methylation in CRC patients. Fourteen studies were included in the meta-analysis after screening 120 articles. The following items were collected from each study: author, published year, country, patient gender, MGMT methylation status, and patients' disease progression. Pooled hazard ratios and odd ratios with 95% confidence intervals (CIs) were calculated using fixed or random effect models depending on the heterogeneity between studies. The overall survival of CRC patients was found not to be significantly associated with MGMT methylation. Further subgroup analysis showed that the frequency of MGMT methylation was significantly higher in CRC than in normal tissues (p MGMT promoter in CRC patients was more frequently methylated than in adenoma patients. In addition, MGMT methylation was significantly increased in adenoma than in normal tissues (p MGMT methylation is central to the development of cancer that involves a stepwise carcinogenesis of normal adenoma carcinoma cascade. However, MGMT methylation is not associated with the prognosis of CRC.

  19. Clinical value of methylation of plasma adenomatous polyposis coli gene in the molecular diagnosis of hepatocellular carcinoma%腺瘤性结肠息肉病基因甲基化在肝细胞癌分子诊断中的价值

    Institute of Scientific and Technical Information of China (English)

    胡瑜; 华东; 程之红; 吴玉玉; 谢其根; 王琼瑶; 余坚; 黄朝晖

    2011-01-01

    Objective: To establish a method of methylation-sensitive restriction enzymes-quantitative PCR (MSRE-Qpcr) for methylation analysis of adenomatous polyposis coli (APC) gene, and to further assess the clinical value of plasma methylation detection by using this method in diagnosis of hepatocellular carcinoma (HCC). Methods: Hha I was used to digest genomic DNA, and the digestion efficiency was evaluated by using Qpcr technique. Then the optimized MSRE-Qpcr method was established. The methylation levels of APC in 45 liver tissues (20 surgically resected HCC specimens and the matched non-cancerous tissues, as well as 5 normal liver tissues) were detected by MSRE-Qpcr, and then further validated by using bisulfite sequencing PCR (BSP). The results of MSRE-Qpcr were compared with those of methylation-specific PCR (MSP) assay. MSRE-Qpcr method was used to detect the APC methylation status of plasma samples from 72 cases of HCC, 37 cases of benign liver diseases and 41 healthy volunteers. Results: The established MSRE-Qpcr method could detect as low as 1% methylated target sites in given DNA samples. The results of MSRE-Qpcr and MSP showed that APC gene was hypermethylated in HCC tissues. The result of MSRE-Qpcr was verified by BSP, and it was comparable with that of MSP (Kappa=0.955, P<0.000 1). Methylation level of plasma APC in patients with HCC was significant higher than those in patients with benign liver diseases and the healthy volunteers (P<0.000 1).Combined analysis of plasma APC methylation and serum alpha-fetoprotein (AFP) revealed an increased diagnostic efficacy for HCC. Conclusion: MSRE-Qpcr is a method for quantitative analysis of APC methylation level. Methylation analysis of plasma APC is a valuable method for the noninvasive diagnosis of HCC.%目的:建立甲基化敏感性限制性内切酶-定量PCR (methylation-sensitive restriction enzyme-quantitative PCR,MSRE-qPCR)方法,并应用该方法探讨血浆腺瘤性结肠息肉病(adenomatous polyposis

  20. The possible role of TP53 mutation status in the treatment of squamous cell carcinomas of the head and neck (HNSCC) with radiotherapy with different overall treatment times

    DEFF Research Database (Denmark)

    Eriksen, Jesper Grau; Alsner, Jan; Steiniche, Torben;

    2005-01-01

    for the outcome of radiotherapy. PATIENTS AND METHODS: DNA extracted from 180 paraffin-embedded formalin-fixed pretreatment biopsies of HNSCC was screened for mutations in exon 4C-10 by denaturing high-pressure liquid chromatography (DHPLC) followed by sequencing. Treatment was 66-68Gy, 2Gy/fx with overall...... treatment times of 6.5 and 5.5 weeks according to the DAHANCA-guidelines. Endpoints were local control at T-site, disease-specific and crude survival. RESULTS: 125 of 180 carcinomas (69%) carried in total 176 mutations. 72 carcinomas were WT (40%) and 108 carcinomas (60%) carried mutations giving...... dysfunctional p53. Overall, mutations in TP53 were not associated with the endpoints. However, when dichotomising according to TP53 status and evaluating the effect of the overall treatment time then tumours with mutant TP53 did benefit from 6 instead of 5fx/wk regarding local control, P=0.005; RR: 0.33 (C.I 95...

  1. Assessment of BRAF V600E Status in Colorectal Carcinoma: Tissue-Specific Discordances between Immunohistochemistry and Sequencing.

    Science.gov (United States)

    Estrella, Jeannelyn S; Tetzlaff, Michael T; Bassett, Roland L; Patel, Keyur P; Williams, Michelle D; Curry, Jonathan L; Rashid, Asif; Hamilton, Stanley R; Broaddus, Russell R

    2015-12-01

    Although sequencing provides the gold standard for identifying colorectal carcinoma with BRAF V600E mutation, immunohistochemistry (IHC) with the recently developed mouse monoclonal antibody VE1 for BRAF V600E protein has shown promise as a more widely available and rapid method. However, we identified anecdotal discordance between VE1 IHC and sequencing results and therefore analyzed VE1 staining by two different IHC methods (Leica Bond and Ventana BenchMark) in whole tissue sections from 480 colorectal carcinomas (323 BRAF wild-type, 142 BRAF V600E mutation, and 15 BRAF non-V600E mutation). We also compared the results with melanomas and papillary thyroid carcinomas (PTC). With the Bond method, among 142 BRAF V600E-mutated colorectal carcinomas, 77 (54%) had diffuse VE1 staining and 48 (33%) had heterogeneous staining, but 17 (12%) were negative. Among 323 BRAF wild-type colorectal carcinomas, 196 (61%) were negative, but 127 (39%) had staining, including 7 with diffuse staining. When positivity was defined as staining in ≥ 20% of tumor cells, VE1 IHC had sensitivity of 75% and specificity of 93% for BRAF V600E mutation. With the Ventana method, among 57 BRAF V600E-mutated colorectal carcinomas, 36 (63%) had diffuse VE1 staining, whereas 6 (11%) had no or weak (Ventana VE1 IHC in melanoma and PTC were highly concordant with sequencing results. We conclude that VE1 IHC produces suboptimal results in colorectal carcinoma and should not be used to guide patient management.

  2. Parenting, Socioeconomic Status Risk, and Later Young Adult Health: Exploration of Opposing Indirect Effects via DNA Methylation

    Science.gov (United States)

    Beach, Steven R. H.; Lei, Man-Kit; Brody, Gene H.; Kim, Sangjin; Barton, Allen W.; Dogan, Meesha V.; Philibert, Robert A.

    2016-01-01

    A sample of 398 African American youth, residing in rural counties with high poverty and unemployment, were followed from ages 11 to 19. Protective parenting was associated with better health, whereas elevated socioeconomic status (SES) risk was associated with poorer health at age 19. Genome-wide epigenetic variation assessed in young adulthood…

  3. Folic acid, polymorphism of methyl-group metabolism genes, and DNA methylation in relation to GI carcinogenesis.

    Science.gov (United States)

    Fang, Jing Yuan; Xiao, Shu Dong

    2003-01-01

    DNA methylation is the main epigenetic modification after replication in humans. DNA (cytosine-5)-methyltransferase (DNMT) catalyzes the transfer of a methyl group from S-adenosyl-L-methionine (SAM) to C5 of cytosine within CpG dinucleotide sequences in the genomic DNA of higher eukaryotes. There is considerable evidence that aberrant DNA methylation plays an integral role in carcinogenesis. Folic acid or folate is crucial for normal DNA synthesis and can regulate DNA methylation, and through this, it affects cellular SAM levels. Folate deficiency results in DNA hypomethylation. Epidemiological studies have indicated that folic acid protects against gastrointestinal (GI) cancers. Methylene-tetrahydrofolate reductase (MTHFR) and methionine synthase (MS) are the enzymes involved in folate metabolism and are thought to influence DNA methylation. MTHFR is highly polymorphic, and the variant genotypes result in decreased MTHFR enzyme activity and lower plasma folate level. Two common MTHFR polymorphisms, 677CT (or 677TT) and A1298C, and an MS polymorphism, A-->G at 2756, have been identified. Most studies support an inverse association between folate status and the rate of colorectal adenomas and carcinomas. During human GI carcinogenesis, MTHFR is highly polymorphic, and the variant genotypes result in decreased MTHFR enzyme activity and lower plasma folate level, as well as aberrant methylation.

  4. Denture polymers with antimicrobial properties: a review of the development and current status of anionic poly(methyl methacrylate) polymers.

    Science.gov (United States)

    Raj, Periathamby Antony; Dentino, Andrew R

    2013-09-01

    The denture base polymer poly(methyl methacrylate) (PMMA) is highly susceptible for microbial colonization resulting in denture-associated infections. Over the years research has focused on ways to modify the PMMA properties via surface and chemical modification. These studies led to the development of new denture polymers that include anionic PMMA polymers. The new anionic polymers presented the possibility of compromising the physical and mechanical properties required for denture fabrication. These obstacles were overcome by generating anionic PMMA polymers with physical and mechanical properties suitable for denture fabrication. A large body of literature is available on the anionic PMMA polymers, their antimicrobial properties and their potential for the commercial and clinical application as dental biomaterials. This article describes a review and evaluation of the anionic PMMA polymers for their suitability to serve as denture base polymers, their antimicrobial properties, their efficacy to prevent denture-induced infection and their safety in the oral environment.

  5. Quantitative Methylation Profiles for Multiple Tumor Suppressor Gene Promoters in Salivary Gland Tumors

    Science.gov (United States)

    Durr, Megan L.; Mydlarz, Wojciech K.; Shao, Chunbo; Zahurak, Marianna L.; Chuang, Alice Y.; Hoque, Mohammad O.; Westra, William H.; Liegeois, Nanette J.; Califano, Joseph A.; Sidransky, David; Ha, Patrick K.

    2010-01-01

    Background Methylation profiling of tumor suppressor gene (TSGs) promoters is quickly becoming a powerful diagnostic tool for the early detection, prognosis, and even prediction of clinical response to treatment. Few studies address this in salivary gland tumors (SGTs); hence the promoter methylation profile of various TSGs was quantitatively assessed in primary SGT tissue to determine if tumor-specific alterations could be detected. Methodology DNA isolated from 78 tumor and 17 normal parotid gland specimens was assayed for promoter methylation status of 19 TSGs by fluorescence-based, quantitative methylation-specific PCR (qMSP). The data were utilized in a binary fashion as well as quantitatively (using a methylation quotient) allowing for better profiling and interpretation of results. Principal Findings The average number of methylation events across the studied genes was highest in salivary duct carcinoma (SDC), with a methylation value of 9.6, compared to the normal 4.5 (p<0.0003). There was a variable frequency and individual methylation quotient detected, depending on the TSG and the tumor type. When comparing normal, benign, and malignant SGTs, there was a statistically significant trend for increasing methylation in APC, Mint 1, PGP9.5, RAR-β, and Timp3. Conclusions/Significance Screening promoter methylation profiles in SGTs showed considerable heterogeneity. The methylation status of certain markers was surprisingly high in even normal salivary tissue, confirming the need for such controls. Several TSGs were found to be associated with malignant SGTs, especially SDC. Further study is needed to evaluate the potential use of these associations in the detection, prognosis, and therapeutic outcome of these rare tumors. PMID:20520817

  6. Levels of DNA Methylation Vary at CpG Sites across the BRCA1 Promoter, and Differ According to Triple Negative and "BRCA-Like" Status, in Both Blood and Tumour DNA.

    Directory of Open Access Journals (Sweden)

    Sarah L Daniels

    Full Text Available Triple negative breast cancer is typically an aggressive and difficult to treat subtype. It is often associated with loss of function of the BRCA1 gene, either through mutation, loss of heterozygosity or methylation. This study aimed to measure methylation of the BRCA1 gene promoter at individual CpG sites in blood, tumour and normal breast tissue, to assess whether levels were correlated between different tissues, and with triple negative receptor status, histopathological scoring for BRCA-like features and BRCA1 protein expression. Blood DNA methylation levels were significantly correlated with tumour methylation at 9 of 11 CpG sites examined (p<0.0007. The levels of tumour DNA methylation were significantly higher in triple negative tumours, and in tumours with high BRCA-like histopathological scores (10 of 11 CpG sites; p<0.01 and p<0.007 respectively. Similar results were observed in blood DNA (6 of 11 CpG sites; p<0.03 and 7 of 11 CpG sites; p<0.02 respectively. This study provides insight into the pattern of CpG methylation across the BRCA1 promoter, and supports previous studies suggesting that tumours with BRCA1 promoter methylation have similar features to those with BRCA1 mutations, and therefore may be suitable for the same targeted therapies.

  7. Frequent Promoter Methylation of CDH1, DAPK, RARB, and HIC1 Genes in Carcinoma of Cervix Uteri: Its Relationship to Clinical Outcome

    Directory of Open Access Journals (Sweden)

    Schneider Achim

    2003-05-01

    Full Text Available Abstract Background Cervical cancer (CC, a leading cause of cancer-related deaths in women worldwide, has been causally linked to genital human papillomavirus (HPV infection. Although a host of genetic alterations have been identified, molecular basis of CC development is still poorly understood. Results We examined the role of promoter hypermethylation, an epigenetic alteration that is associated with the silencing tumor suppressor genes in human cancer, by studying 16 gene promoters in 90 CC cases. We found a high frequency of promoter methylation in CDH1, DAPK, RARB, and HIC1 genes. Correlation of promoter methylation with clinical characteristics and other genetic changes revealed the following: a overall promoter methylation was higher in more advanced stage of the disease, b promoter methylation of RARB and BRCA1 predicted worse prognosis, and c the HIC1 promoter methylation was frequently seen in association with microsatellite instability. Promoter methylation was associated with gene silencing in CC cell lines. Treatment with methylation or histone deacetylation-inhibiting agents resulted in profound reactivation of gene expression. Conclusions These results may have implications in understanding the underlying epigenetic mechanisms in CC development, provide prognostic indicators, and identify important gene targets for treatment.

  8. The EGFR mutation status affects the relative biological effectiveness of carbon-ion beams in non-small cell lung carcinoma cells.

    Science.gov (United States)

    Amornwichet, Napapat; Oike, Takahiro; Shibata, Atsushi; Nirodi, Chaitanya S; Ogiwara, Hideaki; Makino, Haruhiko; Kimura, Yuka; Hirota, Yuka; Isono, Mayu; Yoshida, Yukari; Ohno, Tatsuya; Kohno, Takashi; Nakano, Takashi

    2015-06-11

    Carbon-ion radiotherapy (CIRT) holds promise to treat inoperable locally-advanced non-small cell lung carcinoma (NSCLC), a disease poorly controlled by standard chemoradiotherapy using X-rays. Since CIRT is an extremely limited medical resource, selection of NSCLC patients likely to benefit from it is important; however, biological predictors of response to CIRT are ill-defined. The present study investigated the association between the mutational status of EGFR and KRAS, driver genes frequently mutated in NSCLC, and the relative biological effectiveness (RBE) of carbon-ion beams over X-rays. The assessment of 15 NSCLC lines of different EGFR/KRAS mutational status and that of isogenic NSCLC lines expressing wild-type or mutant EGFR revealed that EGFR-mutant NSCLC cells, but not KRAS-mutant cells, show low RBE. This was attributable to (i) the high X-ray sensitivity of EGFR-mutant cells, since EGFR mutation is associated with a defect in non-homologous end joining, a major pathway for DNA double-strand break (DSB) repair, and (ii) the strong cell-killing effect of carbon-ion beams due to poor repair of carbon-ion beam-induced DSBs regardless of EGFR mutation status. These data highlight the potential of EGFR mutation status as a predictor of response to CIRT, i.e., CIRT may show a high therapeutic index in EGFR mutation-negative NSCLC.

  9. Construction of Differential-Methylation Subtractive Library

    Directory of Open Access Journals (Sweden)

    Wei Hu

    2014-01-01

    Full Text Available Stress-induced ROS changes DNA methylation patterns. A protocol combining methylation-sensitive restriction endonuclease (MS-RE digestion with suppression subtractive hybridization (SSH to construct the differential-methylation subtractive library was developed for finding genes regulated by methylation mechanism under cold stress. The total efficiency of target fragment detection was 74.64%. DNA methylation analysis demonstrated the methylation status of target fragments changed after low temperature or DNA methyltransferase inhibitor treatment. Transcription level analysis indicated that demethylation of DNA promotes gene expression level. The results proved that our protocol was reliable and efficient to obtain gene fragments in differential-methylation status.

  10. Human papillomavirus DNA and p16 expression in Japanese patients with oropharyngeal squamous cell carcinoma

    OpenAIRE

    Kawakami, Hisato; Okamoto, Isamu; Terao, Kyoichi; Sakai, Kazuko; SUZUKI, MINORU; Ueda, Shinya; Tanaka, Kaoru; Kuwata, Kiyoko; Morita, Yume; Ono, Koji; Nishio, Kazuto; Nishimura, Yasumasa; Doi, Katsumi; Nakagawa, Kazuhiko

    2013-01-01

    Human papillomavirus (HPV) is a major etiologic factor for oropharyngeal squamous cell carcinoma (OPSCC). However, little is known about HPV-related OPSCC in Japan. During the study, formalin-fixed, paraffin-embedded OPSCC specimens from Japanese patients were analyzed for HPV DNA by the polymerase chain reaction (PCR) and for the surrogate marker p16 by immuno-histochemistry. For HPV DNA-positive, p16-negative specimens, the methylation status of the p16 gene promoter was examined by methyla...

  11. Hyperplastic polyps of the colon and rectum - reclassification, BRAF and KRAS status in index polyps and subsequent colorectal carcinoma

    DEFF Research Database (Denmark)

    Janjua, Huma Gul Rehana; Høgdall, Estrid; Linnemann, Dorte

    2015-01-01

    in the follow-up period (1 patient had SSL, 4 had THP, and 2 had unspecified non-neoplastic lesions). Ten patients developed other gastrointestinal malignancies. The patient with SSL as index lesions who developed CRC harbored V600E BRAF mutation in both index lesion and the carcinoma. Sixteen percent...

  12. Expression of lysosome-associated protein transmembrane 4B-35 in cancer and its correlation with the differentiation status of hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Cong Peng; Rou-Li Zhou; Gen-Ze Shao; Jing-An Rui; Shao-Bin Wang; Ming Lin; Sha Zhang; Zi-Feng Gao

    2005-01-01

    AIM: To produce high-quality polydonal antibody to lysosome associated protein transmembrane 4B-35 and to identify LAPTM4B-35 expression in cancer tissues and its correlation with differentiation status of hepatocellular carcinoma (HCC).METHODS: The 297 bp 5' end of LAPTM4BcDNA was obtained by PCR and inserted into prokaryotic expression vector pGEXKG. Then the recombinant pGEX-KG-N1-99 was transformedinto E. coli JM109 to express GST-fusion protein. The fusion protein was purified by glutathione sepharoseTM 4B agarose. The purified GST-LAPTM4B-N1-99 was characterized by SDSPAGE, and used to immunize rabbits. The titer and specificity of antisera were detected by ELISA and Western blot, respectively. The correlation between the expression levels of LAPTM4B-35 and the differentiation status of HCC was analyzed via Western blot. The expression of LAPTM4B35 in HCC and other six cancer tissues was investigated via tissue chip and immunohistochemical analysis. RESULTS: About 6.2 mg of pure GST-LAPTM4B-N1-99 was isolated from 1 L of bacteria. The GST-LAPTM4B-N1-99produced high titer antisera in rabbits and showed good immunity. Western blot showed specific reactions for the antibody to the LAPTM4B-35 in the total proteins from HCC tissues and BEL-7402 cells, also to the fusion protein purified or in the transformed bacteria. LAPTM4B-35 was remarkably expressed in several cancers, such as HCC, breast cancer, gastric carcinoma, lung cancer, and colon carcinoma, but not commonly expressed in esophageal cancer and rectum carcinoma. Notably, the expression levels of LAPTM4B-35 were significantly and inversely correlated to the differentiation of HCCs in a 20 case analysis. CONCLUSION: Specific polyclonal antibody (LAPTM4B-N1-99-pAb) to LAPTM4B-35 was produced. It identified the expression of LAPTM4B-35 in some cancer tissues originated from single layer cuboidal and columnar epithelial cells and firmly demonstrated that the expression of LAPTM4B-35 in HCC was inversely

  13. Promoter Hypermethylation of DNA Repair Gene MGMT in Laryngeal Squamous Cell Carcinoma

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    The relationship between hypermethylation of CpG islands in the promoter regions of O6methylguanine DNA methyltransferase (MGMT)genes and laryngeal squamous cell carcinoma was explored. Methylation-specific PCR and semi-quantitative RT-PCR were used to study the promoter methylation and mRNA expression of the MGMT gene in laryngeal carcinoma tissues, t issues adjacent to the tumor and normal laryngeal tissues. Hypermethylation of MGMT gene was detected in 16 samples of 46 (34.8 %) laryngeal squamous cell carcinoma samples. However, the MGMT hypermethylation was not detected in all tissues adjacent to the tumors and normal tissues. No significant difference in MGMT gene hypermethylation was found in samples with different histological grades (x2= 3. 130, P=0. 077) or in samples from patients with different TNM status (x2=3. 957, P=0. 138). No expression of MGMT mRNA was detected in all hypermethylated laryngeal carcinoma tissues. The expression of MGMT mRNA was detected in all unmethylated laryngeal carcinoma tissues, tissues adjacent to the tumors and normal tissues. It suggests that MGMT gene promoter hypermethylation is associated with MGMT gene transcription loss in laryngeal carcinoma tissues and possibly plays an important role in carcinogenesis of laryngeal tissues.

  14. Comparing the expression of myoepithelial cell markers CD10 and smooth muscle actin with the estrogen receptor status in the invasive carcinoma breast: An immunohistochemical study

    Directory of Open Access Journals (Sweden)

    Gaurav Arora

    2013-01-01

    Full Text Available Background: Breast cancer is the most frequent cancer in females throughout the world. Around 20% of breast carcinomas are estrogen receptor alpha (ER-negative. Thus, theoretically this negativity could be either the result of down-regulation of ER expression in the tumor cells, or the result of the tumor being derived from cells which normally lack that expression. Normal basal, including myoepithelial cells of the breast is ER-negative. CD10 and smooth muscle actin (SMA are used as markers for the demonstration of these basal cells. Aims: To compare the expression of positive staining for CD10 and SMA in ER-negative and ER-positive invasive breast carcinomas. Materials and Methods: The study was performed on 40 paraffin-embedded tissues of already diagnosed cases of invasive breast carcinomas with known ER status, i.e., thirty ER-negative and ten ER-positive cases. Expression of CD10 and SMA was demonstrated using avidin-biotin-peroxidase complex (ABC technique. Tumor was considered to be positive for both markers only when more than 10% of tumor cells were stained positive. Results: Overall, CD10 tumor cell staining was seen in eight, 23.3% (7/30 ER-negative cases and in 10% (1/10 ER-positive cases. Also the staining intensity was considered to be strong. SMA tumor cell staining was seen in only 6.7% (2/30 ER-negative cases and the staining intensity was considered to be moderate. Percentages of positively stained tumor cells varied between 13% to 72% and 23% to 45% for CD10 and SMA, respectively. Conclusion: CD10 is a better marker when compared to SMA, as it is expressed in more number of cases and gives strong positivity in tumor cells. Higher expression of CD10 and SMA is correlated with higher tumor grade and ER negativity.

  15. Genome-wide methylation profiling and the PI3K-AKT pathway analysis associated with smoking in urothelial cell carcinoma

    NARCIS (Netherlands)

    Brait, Mariana; Munari, Enrico; LeBron, Cynthia; Noordhuis, Maartje G.; Begum, Shahnaz; Michailidi, Christina; Gonzalez-Roibon, Nilda; Maldonado, Leonel; Sen, Tanusree; Guerrero-Preston, Rafael; Cope, Leslie; Parrella, Paola; Fazio, VitoMichele; Ha, Patrick K.; Netto, George J.; Sidransky, David; Hoque, Mohammad O.

    2013-01-01

    Urothelial cell carcinoma (UCC) is the second most common genitourinary malignant disease in the USA, and tobacco smoking is the major known risk factor for UCC development. Exposure to carcinogens, such as those contained in tobacco smoke, is known to directly or indirectly damage DNA, causing muta

  16. Identification of candidate methylation-responsive genes in ovarian cancer

    Directory of Open Access Journals (Sweden)

    Dickerson Erin B

    2007-01-01

    Full Text Available Abstract Background Aberrant methylation of gene promoter regions has been linked to changes in gene expression in cancer development and progression. Genes associated with CpG islands (CGIs are especially prone to methylation, but not all CGI-associated genes display changes in methylation patterns in cancers. Results In order to identify genes subject to regulation by methylation, we conducted gene expression profile analyses of an ovarian cancer cell line (OVCAR-3 before and after treatment with the demethylating agent 5-aza-deoxycytidine (5-aza-dC. An overlapping subset of these genes was found to display significant differences in gene expression between normal ovarian surface epithelial cells and malignant cells isolated from ovarian carcinomas. While 40% of all human genes are associated with CGIs, > 94% of the overlapping subset of genes is associated with CGIs. The predicted change in methylation status of genes randomly selected from the overlapping subset was experimentally verified. Conclusion We conclude that correlating genes that are upregulated in response to 5-aza-dC treatment of cancer cell lines with genes that are down-regulated in cancer cells may be a useful method to identify genes experiencing epigenetic-mediated changes in expression over cancer development.

  17. Energy and protein intake and nutritional status in non-surgically treated patients with small cell anaplastic carcinoma of the lung

    Energy Technology Data Exchange (ETDEWEB)

    Enig, B.; Winther, E.; Hessov, I.

    The spontaneous food intake and nutritional status was assessed in 23 patients with small cell anaplastic carcinoma of the lung before and two times during a treatment period of 6 weeks. Radiation therapy was given for 2 weeks followed by a course of chemotherapy and another 2 weeks of radiation therapy. The energy intake decreased during the treatment from 146 to 130 per cent of basal metabolic rate. The protein intake remained unchanged (mean 0.9 g/kg body weight). There were insignificant and small losses of weight, body fat, free body mass and arm muscle circumference, and no changes were seen in serum albumin and serum transferrin. However, 6 patients suffered a weight loss of 5 per cent or more. No correlation existed between the nutritional parameters measured before treatment and the changes during treatment. Patients who suffered a loss of body weight could therefore not be singled out before the treatment.

  18. Parkin基因甲基化对鼻咽癌早期诊断及预后判断的价值研究%Parkin gene promoter methylation in nasopharyngeal carcinoma

    Institute of Scientific and Technical Information of China (English)

    江波; 倪海峰; 周珍; 李勇; 黄光武

    2016-01-01

    目的:研究Parkin基因甲基化在鼻咽癌早期诊断及预后判断中的价值。方法运用甲基化特异性PCR检测54例鼻咽癌组织、16例慢性鼻咽炎症组织和16例正常鼻咽上皮组织中Parkin基因启动子区甲基化情况,并分析其与鼻咽癌临床生物学因素的关系。结果慢性鼻咽炎症组织和正常鼻咽上皮组织中均未检测到Parkin基因甲基化,而在鼻咽癌组织中甲基化率为62.96%(34/54),3种鼻咽组织Parkin基因甲基化情况比较有统计学差异(P<0.05)。进一步统计学分析发现鼻咽癌组织Parkin基因甲基化与其临床生物学因素均无关(均P>0.05)。结论 Parkin基因甲基化具有肿瘤特异性,与鼻咽癌发生、发展密切相关,有望成为早期分子生物学辅助诊断的标志物,但不能作为判断鼻咽癌预后的预测指标。%Objective To investigate Parkin gene promoter methylation in nasopharyngeal carcinoma (NPC). Methods The methylation- specific PCR (MSP) was used to detect methylation level of Parkin gene in 54 tissue specimens of NPC, 16 specimens of chronic nasopharyngitis and 16 specimens of normal nasopharyngeal epithelia tissues, and its relationship with the clinical and biological features of NPC was analyzed. Results Parkin gene promoter methylation was detected in 62.96%(34/54) of NPC specimens, not in any specimens of chronic nasopharyngitis and normal nasopharyngeal epithelia tissues(P0.05). Conclusion Parkin gene promoter methylation has high specificity in distinguishing NPC from normal nasopharyngeal tissues and inflammatory nasopharyngeal disease, suggesting that it may be used for early molecular diagnosis of NPC.

  19. Expression status of CD44 and CD133 as a prognostic marker in esophageal squamous cell carcinoma treated with neoadjuvant chemotherapy followed by radical esophagectomy.

    Science.gov (United States)

    Okamoto, Koichi; Ninomiya, Itasu; Ohbatake, Yoshinao; Hirose, Atsushi; Tsukada, Tomoya; Nakanuma, Shinichi; Sakai, Seisho; Kinoshita, Jun; Makino, Isamu; Nakamura, Keishi; Hayashi, Hironori; Oyama, Katsunobu; Inokuchi, Masafumi; Nakagawara, Hisatoshi; Miyashita, Tomoharu; Hidehiro, Tajima; Takamura, Hiroyuki; Fushida, Sachio; Ohta, Tetsuo

    2016-12-01

    Cancer stem cells (CSCs) have self-renewal and pluripotency capabilities and contribute to cancer progression and chemoresistance. It has been proposed that the treatment resistance and heterogeneity of CSCs are deeply involved in the prognosis of patients with esophageal squamous cell carcinoma (ESCC). The objective of this study was to identify the influence of the expression status of the CSC markers CD44 and CD133 on chemotherapeutic efficacy and prognosis in ESCC patients who underwent radical esophagectomy after neoadjuvant chemotherapy (NAC). Endoscopically biopsied specimens taken before NAC and surgically resected specimens after NAC were immunohistochemically assessed for CD44 and CD133 expression for 47 ESCC patients who underwent NAC followed by radical esophagectomy. The correlation between CD44 and CD133 expression status and clinicopathological findings and the prognosis of ESCC patients after NAC followed by esophagectomy were analyzed. The percentages of CD44-positive cells and CD133-positive cells in specimens were increased after NAC. CD44 and CD133 expression status before NAC did not correlate with the degree of tumor progression and had no impact on the chemotherapeutic effect. However, strong expression of CD44 or CD133 and a high proportion of CD133-expressing cells before NAC were significantly associated with poorer esophageal cancer-specific survival. Patients with strong expression of CD44 or CD133 and those with a high ratio of CD133-positive tumor cells showed significantly poor prognosis regardless of the effect of chemotherapy. Multivariate analysis showed that simultaneous strong expression of CD44 and CD133 before NAC, a high rate of CD133-positive tumor cells before NAC, and primary tumor remission assessed by preoperative endoscopy were significant independent prognostic factors for ESCC. Our data indicate that CD44 and CD133 expression status prior to treatment dictates the malignant potential of ESCC and may be a novel

  20. Androgen receptor gene methylation and exon one CAG repeat length in ovarian cancer: differences from breast cancer.

    Science.gov (United States)

    Kassim, Samar; Zoheiry, Nivan M; Hamed, Wael M; Going, James J; Craft, John A

    2004-07-01

    More than one neoplastic founder clone can exist in benign epithelial tumours. Although theories of clonal selection make pluriclonality appear unlikely in carcinomas, published data do not exclude this possibility. This study looked for evidence of multiclonal X inactivation in ovarian carcinoma using AR methylation as a marker. Fifteen unifocal ovarian carcinomas and 14 multifocal carcinomas all in Scottish patients were studied. One representative formalin-fixed paraffin-embedded tumour block was chosen for each of the former and two for the latter. From each of these 43 tumour blocks three samples each of approximately 10(4) carcinoma cells were obtained by microdissection (129 in all). DNA released by proteinase K digestion was subjected to PCR amplification of the androgen receptor gene AR exon I CAG repeat polymorphism with and without prior digestion with methylation-sensitive restriction enzymes HpaII and HhaI. Complex amplification patterns were consistent with mosaic X inactivation in some ovarian carcinomas but acquired anomalies of AR methylation cannot be excluded. Parallel analysis of other X-linked polymorphic loci would strengthen the inference of clonality status from DNA methylation data in tumour X studies. Strikingly, the number of CAG repeats in the 29 ovarian tumour patients (median 16, range 11 - 20) was substantially fewer than in 34 previously studied breast cancer patients from the same scottish population (median 21, range 14 - 26; P CAG repeat were over-represented in the ovarian cancer patients but not in the breast cancer series. These findings reinforce recent suggestions that AR may have a role in ovarian carcinogenesis.

  1. Relationship between PTEN, DNA mismatch repair, and tumor histotype in endometrial carcinoma: retained positive expression of PTEN preferentially identifies sporadic non-endometrioid carcinomas.

    Science.gov (United States)

    Djordjevic, Bojana; Barkoh, Bedia A; Luthra, Rajyalakshmi; Broaddus, Russell R

    2013-10-01

    Loss of PTEN (phosphatase and tensin homolog) expression and microsatellite instability are two of the more common molecular alterations in endometrial carcinoma. From the published literature, it is controversial as to whether there is a relationship between these different molecular mechanisms. Therefore, a cohort of 187 pure endometrioid and non-endometrioid endometrial carcinomas, carefully characterized as to clinical and pathological features, was examined for PTEN sequence abnormalities and the immunohistochemical expression of PTEN and the DNA mismatch repair proteins MLH1, MSH2, MSH6, and PMS2. MLH1 methylation analysis was performed when tumors had loss of MLH1 protein. Mismatch repair protein loss was more frequent in endometrioid carcinomas compared with non-endometrioid carcinomas, a difference primarily attributable to the presence of MLH1 methylation in a greater proportion of endometrioid tumors. Among the non-endometrioid group, mixed endometrioid/non-endometrioid carcinomas were the histotype that most commonly had loss of a mismatch repair protein. In endometrioid tumors, the frequency of PTEN loss measured by immunohistochemistry and mutation did not differ significantly between the mismatch repair protein intact or mismatch repair protein loss groups, suggesting that PTEN loss is independent of mismatch protein repair status in this group. However, in non-endometrioid carcinomas, both intact positive PTEN immunohistochemical expression and PTEN wild type were highly associated with retained positive expression of mismatch repair proteins in the tumor. Relevant to screening endometrial cancers for Lynch Syndrome, an initial PTEN immunohistochemistry determination may be able to replace the use of four mismatch repair immunohistochemical markers in 63% of patients with non-endometrioid endometrial carcinoma. Therefore, PTEN immunohistochemistry, in combination with tumor histotype, is a useful adjunct in the clinical evaluation of endometrial

  2. Methylenetetrahydrofolate reductase C677T genotype affects promoter methylation of tumor-specific genes in sporadic colorectal cancer through an interaction with folate/vitamin B12 status

    Institute of Scientific and Technical Information of China (English)

    Pooneh Mokarram; Fakhraddin Naghibalhossaini; Mehdi Saberi Firoozi; Seyed Vahid Hosseini; Ahmad Izadpanah; Heshmetalah Salahi; Seyed Ali Malek-Hosseini; Abdoulrasool Talei; Mehra Mojallal

    2008-01-01

    AIM: To evaluate joint effects of Methy/entetra-hydrofolate reductase (MTHFR) C677Tgenotypes, and serum folate/vitamin B12 concentrations on promoter methylation of tumor-associated genes among Iranian colorectal cancer patients.METHODS: We examined the associations between MTHFR C677T genotype, and promoter methylation of P16, Hmlh1, and Hmsh2 tumor-related genes amonq 151 sporadic colorectal cancer patients. The promoter methylation of tumor-related genes was determined by methylation-specific PCR. Eighty six patients from whom fresh tumor samples were obtained and 81 controls were also examined for serum folate and vitamin B12, concentrations by a commercia radioimmunoassay kit.RESULTS: We found 29.1% of cases had tumors with at least one methylated gene promoter. In case-case comparison, we did not find a significant association between methylation in tumors and any single genotype. However, in comparison to controls with the CC genotype, an increased risk of tumor methylation was associated with the CT genotype (OR=2.5;95% CI,1.1-5.6). In case-case comparisons, folate/vitamin B12 levels were positively associated with tumor methylation. Adjusted odds ratios for tumor methylation in cases with high (above median) versus low (below median) serum folate/vitamin B12 levels were 4.9 (95% CI,1.4-17.7), and 3.9 (95% CI,1.1-13.9), respectively. The frequency of methylated tumors was significantly higher in high methyl donor than low methyl donor group, especially in those with MTHFR CT (P=0.01), and CT/TT (P=0.002) genotypes, but not in those with the CC genotype (P=1.0).CONCLUSION: We conclude that high concentrations of serum folate/vitamin B12 levels are associated with the risk of promoter methylation in tumor-specific genes, and this relationship is modified by MTHFR C677T genotypes.

  3. Methylation status of the SOCS 1 and JUNB genes in chronic myeloid leukemia patients Padrão de metilação dos genes SOCS 1 e JUNB em pacientes com leucemia mieloide crônica

    Directory of Open Access Journals (Sweden)

    Márcia Cristina R. Pena

    2009-01-01

    Full Text Available Alterations in the methylation status of genes may contribute to the progression of Chronic Myeloid Leukemia (CML. In this study, the methylation status in exon2 of SOCS- 1 and promoter regions of both SOCS- 1 and JUNB were evaluated in CML patients. The methylation status of these genes was analyzed using methylation- specific Polymerase Chain Reaction (MSP in 30 samples from CML patients, 30 samples from these same patients after hematopoietic stem cell transplantation (HSCT and 30 samples from healthy controls. The samples of CML patients presented methylation as follows: JUNB gene (3.3%, promoter region of the SOCS- 1 gene (6.6% and exon2 of the SOCS- 1 gene (46.6%. The samples of the healthy individuals presented methylation (10%, P = 0.002 only in exon 2 of the SOCS- 1 gene. After transplantation, patients presented alterations in the methylation status of the promoter region of the SOCS- 1 gene (6.6%, exon2 of SOCS- 1 (46.6% and the promoter region of the JUNB gene (16.6%. Methylation of the promoter regions of the SOCS- 1 gene and the JUNB gene is not a frequent event in CML. In contrast, SOCS- 1 gene methylation in exon2 is a frequent event, susceptible to alterations in status after HSCT with possible implications for the progression of this disease.Alteração no padrão de metilação gênica pode contribuir para a progressão da leucemia mielóide crônica (LMC. Neste estudo, o padrão de metilação no exon 2 do gene SOCS- 1 e região promotora de ambos SOCS- 1 e JUNB foram avaliadas em pacientes com LMC. O padrão de metilação desses genes foi analisado usando a técnica " methylation- specific polymerase chain reaction (MSP" em 30 amostras de pacientes com LMC, 30 amostras desses mesmos pacientes após transplante de medula óssea (TMO e 30 amostras controle de indivíduos saudáveis. As amostras de pacientes com LMC apresentaram o seguinte padrão de metilação: gene JUNB (3.3%, região promotora do gene SOCS- 1 (6.6% e exon2

  4. Expression of Tenascin C, EGFR, E-Cadherin, and TTF-1 in Medullary Thyroid Carcinoma and the Correlation with RET Mutation Status.

    Science.gov (United States)

    Steiner, Florian; Hauser-Kronberger, Cornelia; Rendl, Gundula; Rodrigues, Margarida; Pirich, Christian

    2016-07-09

    Tenascin C expression correlates with tumor grade and indicates worse prognosis in several tumors. Epidermal growth factor receptor (EGFR) plays an important role in driving proliferation in many tumors. Loss of E-cadherin function is associated with tumor invasion and metastasis. Thyroid transcription factor-1 (TTF-1) is involved in rearranged during transfection (RET) transcription in Hirschsprung's disease. Tenascin C, EGFR, E-cadherin, TTF-1-expression, and their correlations with RET mutation status were investigated in 30 patients with medullary thyroid carcinoma (MTC) (n = 26) or C-cell hyperplasia (n = 4). Tenascin C was found in all, EGFR in 4/26, E-cadherin in 23/26, and TTF-1 in 25/26 MTC. Tenascin C correlated significantly with tumor proliferation (overall, r = 0.61, p C, E-cadherin, and EGFR was r = -0.10, 0.37, and 0.21, respectively. In conclusion, MTC express tenascin C, E-cadherin, and TTF-1. Tenascin C correlates significantly with tumor proliferation, especially in RET-mutated tumors. EGFR is low, and tumors expressing EGFR do not exhibit higher proliferation. TTF-1 does not correlate with RET mutation status and has a weak correlation with tenascin C, E-cadherin, and EGFR expression.

  5. Determination of HER-2 status on FNAC material from breast carcinomas using in situ hybridization with dual chromogen visualization with silver enhancement (dual SISH

    Directory of Open Access Journals (Sweden)

    Beraki Elsa

    2010-01-01

    Full Text Available During the last years, HER-2 status kits and protocols for chromogen visualization of hybridization signals have come on the market. The first generation using chromogen visualization used single color probes. The second generation, now emerging on the market, uses dual chromogen visualization. The aim of this study has been to test a new dual color chromogen kit (Ventana INFORM HER2 Dual Colour ISH Roche ® and compare the results with our in-house method(s. The material consisted primarily of cytological material from invasive breast carcinomas in 49 women. Dual SISH was done on all 49 cytological and histological specimens. The histological specimens were treated according to the manufacturer′s recommendations. The procedure was modified in several steps in order to adapt it to the cytological material. Hybridization failed in two cytological specimens. Dual SISH showed concordant results on cytological and histological material as to amplified/not amplified. The included cases had the same HER-2 expression in the invasive and the in situ components on histology. Four IDC showed HER-2 amplification (8.5%. Polysomy was found in two cases. All dual SISH results except for one concurred with the results of the in-house method(s (1/47=2.1%. The dual SISH is suitable for cytological examination of HER-2 status. The protocol must be optimized for cytological material.

  6. Modulation of lipid peroxidation and antioxidant status upon administration of 'Shemamruthaa' in 7,12-dimethylbenz[a]anthracene induced mammary carcinoma bearing rats

    Institute of Scientific and Technical Information of China (English)

    Ayyakkannu Purushothaman; Elumalai Nandhakumar; Panchanatham Sachdanandam

    2012-01-01

    Objective: To investigate the therapeutic efficacy of a Shemamruthaa (SM), (combination of Hibiscus rosasinensis (H. rosasinensis) flowers, fruits of Phyllanthus emblica (P. emblica) and pure honey in definite ratio), against lipid peroxidation (LPO) and antioxidant status in experimentally induced mammary carcinoma rats. Methods: Adult female Sprague-Dawley rats were used for the study and were divided into four groups. Group I control animals received standard pellet diet and water ad libitum. Group II rats were induced with 7,12-dimethyl benz[a]anthracene (DMBA) (25 mg in 1 mL olive oil) by gastric intubation, whereas another set of DMBA-induced rats were treated with SM (400 mg/kg body weight/d) in olive oil orally by gastric intubation for 14 d after 3 months of induction period (group III). Group IV rats served as SM-treated control animals. At the end of the experimental period, the rats were anaesthetised and sacrificed and used for biochemical measures and histology studies. Results: The LPO was increased and antioxidant levels were decreased in the serum, liver and mammary tissues of cancer-induced rats. The administration of SM drug significantly (P<0.05) decreased LPO and reversed the status of antioxidants to near normal level in cancer-bearing animals. Conclusions: The results obtained indicate the additive and synergistic action of constituents’ plants in the SM drug against oxidative damage and its protective role in DMBA induced mammary cancer.

  7. Detection and Clinical Significance of Methylation of Peripheral Plasma DNA in Patients with Hepatocellular Carcinoma%肝细胞肝癌中外周血浆DNA基因甲基化检测及意义

    Institute of Scientific and Technical Information of China (English)

    王平; 耿小平; 朱立新; 李晓明; 李栋

    2012-01-01

    To evaluate the effect of methylation determination about the peripheral plasma DNA in diagnose of hepatocellular carcinoma (HCC) and select the highly sensitive and specific methylated cancer suppressor genes. Methods Methylation-specific PCR (MSP) was used to detect the degree of methylation about SLIT2 and DAPK genes in peripheral plasma and associated cancer tissues of 34 patients with HCC confirmed by pathology, then analyzed their relationship to clinicopathologic feature. Results The positive rate of the promoter methylation of SLIT2 and DAPK genes in cancer tissues in 34 cases were 70. 6% (24/34) and 79. 4% (27/34) , while the relevant promoter methylation rate in plasma were 44. 1% (15/34) and 50. 0% (17/34) correspondingly. The sensitivity of detection of DNA methylation about SLIT2 and DAPK genes in plasma was 62. 5% and 63. 0%, respectively; both of the specificity for them were 100%. The negative predicted value was 52. 6% and 41. 2%, respectively; while both of the positive predicted value were 100%. There were no significant correlation between the clinicopathologic features and the methylation rate in cancer tissues and plasma (P>0. 05). In plasma of patients whose AFP < 400 μg/L, the positive rate of combined detection of DNA methylation of SLIT2 and DAPK was 61. 1% (11 /18) . Conclusions The detection rate of DNA methylation of SLIT2 and DAPK genes in plasma is higher, and there is a significant correlation between the DNA methylation in HCC tissue and plasma, based on MSP method. DNA methylation in plasma, as an non-invasive method, could be used to diagnose HCC, especially for the patients whose AFP is negative. HBV infection may be only associate with DNA methylation of part gene.%目的 评估外周血浆DNA甲基化检测在肝细胞肝癌(HCC)诊断中的作用,筛选血浆灵敏度及特异性高的甲基化抑癌基因.方法 采用甲基化特异性PCR (MSP)检测34例经病理检查证实的HCC患者血浆

  8. The influence of p53 mutation status on the anti-cancer effect of cisplatin in oral squamous cell carcinoma cell lines

    Science.gov (United States)

    2016-01-01

    Objectives The purpose of this study was to evaluate the anti-cancer activity of cisplatin by studying its effects on cell viability and identifying the mechanisms underlying the induction of cell cycle arrest and apoptosis on oral squamous cell carcinoma (OSCC) cell lines with varying p53 mutation status. Materials and Methods Three OSCC cell lines, YD-8 (p53 point mutation), YD-9 (p53 wild type), and YD-38 (p53 deletion) were used. To determine the cytotoxic effect of cisplatin, MTS assay was performed. The cell cycle alteration and apoptosis were analyzed using flow cytometry. Western blot analysis was used to detect the expression of cell cycle alteration- or apoptosis-related proteins as well as p53. Results Cisplatin showed a time- and dose-dependent anti-proliferative effect in all cell lines. Cisplatin induced G2/M cell accumulation in the three cell lines after treatment with 0.5 and 1.0 µg/mL of cisplatin for 48 hours. The proportion of annexin V-FITC-stained cells increased following treatment with cisplatin. The apoptotic proportion was lower in the YD-38 cell line than in the YD-9 or YD-8 cell lines. Also, immunoblotting analysis indicated that p53 and p21 were detected only in YD-8 and YD-9 cell lines after cisplatin treatment. Conclusion In this study, cisplatin showed anti-cancer effects via G2/M phase arrest and apoptosis, with some difference among OSCC cell lines. The mutation status of p53 might have influenced the difference observed among cell lines. Further studies on p53 mutation status are needed to understand the biological behavior and characteristics of OSCCs and to establish appropriate treatment. PMID:28053903

  9. 结肠癌细胞CDH1基因启动子甲基化对E-上皮钙黏素和β-连接素表达的调控作用%Regulation of CDH1 gene promoter methylation on expression of E-cadherin and β-catenin in human colon carcinoma cells

    Institute of Scientific and Technical Information of China (English)

    李臣; 董坚; 任俊宇; 洪敏; 李少避; 刘为青; 高嫦娥; 陈圣雄; 周华华; 陈明清

    2011-01-01

    Objective To observe the effects of CDH1 gene promoter methylation on the expression of E-cadherin and β-catenin in human colon carcinoma cells.Methods Methylation specific PCR (MSP) and reverse transcription-polymerase chain reaction (RT-PCR) methods were utilized to examine methylation status of CDH1 gene promoter and the changes of E-cadherin mRNA in colon carcinoma cell line HT-29 before and after the treatment with 5-Aza-CdR.The expression of E-cadherin and localization of β-catenin were labeled by immunofluorescence and analyzed under a laser scanning confocal microscope.Results (1) CDH1 gene promoter methylation was positive and unmethylation was negative in HT-29cells before the treatment with 5-Aza-CdR.After treatment with 5-Aza-CdR for 24 h,methylation turned negative and unmethylation was detected; (2) The E-cadherin mRNA failed to be amplified in cells,whereas it could be detected after the treatment.The mRNA expression level of E-cadherin expressed as average gray ratio was 0.491 ±0.011 and 0.568 ±0.013 respectively after treatment with 1 and 2 μmol/L 5-Aza-CdR ( P < 0.05 ; (3) Before treatment with 5-Aza-CdR,the expression of E-cadherin was not detected and β-catenin was detected in cytoplasm and nucleus,while both E-cadherin and β-catenin staining was positive on the cell membrane by immunofluorescence after the treatment.With the expression of E-cadherin induced by 5-Aza-CdR,the membranous expression of β-catenin was elevated and nuclear expression of β-catenin was reduced.Immunofuorescence double staining displayed the distribution of β-catenin was corresponded with E-cadherin on the cell membrane.Conclusion CDH1 gene promoter methylation may lead to the loss of E-cadherin expression and the altered distribution of β-catenin in colon carcinoma cells.%目的 观察结肠癌细胞中上皮钙黏素基因( CDH1)启动子甲基化对上皮钙黏素(E-cadherin)和β-连接素(β-catenin)表达的影响.方法 通过甲基化特异性PCR

  10. Homogeneous MGMT immunoreactivity correlates with an unmethylated MGMT promoter status in brain metastases of various solid tumors.

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    Barbara Ingold

    Full Text Available The O(6-methylguanine-methyltransferase (MGMT promoter methylation status is a predictive parameter for the response of malignant gliomas to alkylating agents such as temozolomide. First clinical reports on treating brain metastases with temozolomide describe varying effects. This may be due to the fact that MGMT promoter methylation of brain metastases has not yet been explored in depth. Therefore, we assessed MGMT promoter methylation of various brain metastases including those derived from lung (n = 91, breast (n = 72 kidney (n = 49 and from malignant melanomas (n = 113 by methylation-specific polymerase chain reaction (MS-PCR and MGMT immunoreactivity. Fifty-nine of 199 brain metastases (29.6% revealed a methylated MGMT promoter. The methylation rate was the highest in brain metastases derived from lung carcinomas (46.5% followed by those from breast carcinoma (28.8%, malignant melanoma (24.7% and from renal carcinoma (20%. A significant correlation of homogeneous MGMT-immunoreactivity (>95% MGMT positive tumor cells and an unmethylated MGMT promoter was found. Promoter methylation was detected in 26 of 61 (43% tumors lacking MGMT immunoreactivity, in 17 of 63 (27% metastases with heterogeneous MGMT expression, but only in 5 of 54 brain metastases (9% showing a homogeneous MGMT immunoreactivity. Our results demonstrate that a significant number of brain metastases reveal a methylated MGMT-promoter. Based on an obvious correlation between homogeneous MGMT immunoreactivity and unmethylated MGMT promoter, we hypothesize that immunohistochemistry for MGMT may be a helpful diagnostic tool to identify those tumors that probably will not benefit from the use of alkylating agents. The discrepancy between promoter methylation and a lack of MGMT immunoreactivity argues for assessing MGMT promoter methylation both by immunohistochemical as well as by molecular approaches for diagnostic purposes.

  11. 肝细胞癌多基因启动子甲基化与预后关系的研究%THE RELATIONSHIP BETWEEN ABERRANT PROMOTER METHYLATION OF MULTIPLE GENES AND PROGNOSIS IN HEPATOCELLULAR CARCINOMA

    Institute of Scientific and Technical Information of China (English)

    彭晓春; 耿小平; 朱立新; 孙昀; 李晓明

    2011-01-01

    To study the relationship between the gene promoter methylation state of DAPK, FHIT and SLIT2 genes and the clinical pronosis of patient in hepatocellular carcinoma ( HCC). Methods The technique of methylation-specific PCR ( MSP) was adopted to investigate the promoter hypermethylalion of DAPK,FHIT 及 SLIT2 genes in 50 HCCs after a curative resection. The relationship between the frequency of hypermethylation of the genes and tumor recrudescence data was analyzed. Results In all patients with HCC, the frequency of hypermethylation in DAPK ,FHIT and SLIT2 were 82. 0% , 68. 0% and 54. 0% , respectively. We divided all those cases into two groups according to the follow-up neoplasm recurrence results( group I ; the group of one year without tumor recrudescence, group Ⅱ : the group of less than one year with tumor recrudescence) . In group I , the frequency of hypermethyla tion in DAPK,FHIT and SLIT2 were 76. 9% , 53. 8% , 50. 0% , respectively; in group II they were 87. 5% , 83. 3% , 58. 3% , re spectively. Those three genes have higer frequency among group D , The frequency hypermethylation of FHIT gene is especially higher in group II (P = 0. 036). In group Ⅱ , there is twenty-two cases which have two or three genes hypermethylation; and in group Ⅰ, the cases are fifteen. There is a statistical prognosis difference between them ( P = 0. 006 ) . Conclusions Hypermethylation of multiple gene promotors are common events in HCC. In patients with HCC, aberrant DNA methylation is significantly associated with poor prog nosis. FHIT maybe can serve as a biomarker for the prognosis, after a curative resection.%目的 了解肝细胞癌(hepatucellular carcinoma,HCC)中,DAPK、FHIT及SLIT2基因的甲基化状况与病人生存预后的关系.方法 应用甲基化特异性PCR(methylation- specific PCR,MSP)技术,检测50例HCC组织中上述基因启动子区域的甲基化状况,并分析每种基因甲基化情况和肿瘤复发之间相关性.结果 50

  12. 5-Aza-CdR对人食管鳞癌Kyse-140细胞BNIP3基因甲基化生物学意义的研究%Biological significance of 5-Aza-CdR on methylation of BNIP3 gene in human esophageal carcinoma cell line Kyse-140

    Institute of Scientific and Technical Information of China (English)

    张洪典; 陈传贵; 胡珊珊; 马钊; 于振涛

    2013-01-01

    -CdR) on the transcriptional regulation through methylation of Bcl-2/adenovirus E1B 19 kd-interacting protein 3 (BNIP3) gene promoter region in human esophageal carcinoma cell line Kyse-140 and its effect on the abilities of growth and proliferation the cell line.METHODS:Human esophageal carcinoma cell line Kyse-140 were cultured and treated with different concentrations of 5-Aza-CdR.Morphological changes of cells were observed under inverted microscope and the proliferative ability of the cell was evaluated by MTT assay.Methylation specific PCR (MSP),RT-PCR and Western blotting were used to detect CpG islands methylation status,the expression of BNIP3 mRNA and protein in Kyse-140 cell line before and after treating with different concentrations of 5-Aza-CdR.RESULTS:Being treated with 1 × 10-7,5 ×10-7,2× 10-6,1 × 10-5 and 5 × 10-5 mol/L 5-Aza-CdR,the inhibition rates of 24 hours were 6.2%,8.4%,11.3%,15.8 % and 26.0 %,the inhibition rates of 48 hours were 8.1%,15.2 %,20.4 %,24.6 % and 39.0 %,and the inhibition rates of 72 hours were 9.3 %,20.5 %,30.1%,40.9 % and 51.0 %.After treated with 5-Aza-CdR,the inhibit ability of tumor cells proliferation was found,and the trend was concentration-dependent(F=18.211,P<0.001) and time-dependent (F=15.411,P=0.002).Hypermethylation status of BNIP3 gene promoter region was observed in Kyse-140 cell line and no expression of the mRNA and protein of BNIP3 gene was found.5-Aza-CdR could reverse BNIP3 gene methylation in Kyse-140 cells and strongly up-regulate the expression of BNIP3 mRNA and protein.CONCLUSIONS:5-Aza-CdR can reverse the methylation status of BNIP3 gene in human esophageal carcionma cell line Kyse-140,which would remove the gene silencing caused by high methylation and thus restore the expression the BNIP3 mRNA and protein levels.It can effectively inhibit growth and proliferation of esophageal cancer of esophageal cancer cells.

  13. Differential methylation status of IGF2-H19 locus does not affect the fertility of crossbred bulls but some of the CTCF binding sites could be potentially important.

    Science.gov (United States)

    Jena, Subas C; Kumar, Sandeep; Rajput, Sandeep; Roy, Bhaskar; Verma, Arpana; Kumaresan, Arumugam; Mohanty, Tushar K; De, Sachinandan; Kumar, Rakesh; Datta, Tirtha K

    2014-04-01

    Associations between abnormal methylation of spermatozoan DNA with male infertility have been sought in recent years to identify a molecular explanation of differential spermatozoan function. The present work was undertaken to investigate the methylation profile of differentially methylated regions (DMRs) in the IGF2-H19 locus of Bos taurus X Bos indicus crossbred bull spermatozoa. Bulls having more than at least 100 insemination records over a period of 12 years were classified into two groups of five bulls each belonging to low- and high-fertility groups. The IGF2 and H19 DMR sequences in B. indicus cattle were observed to be in absolute homology with B. taurus cattle. The DNA of crossbred bull spermatozoa was isolated, bisulfite treated, and amplified for specific DMR regions using methylation-change-specific primers. The overall degree of methylation at IGF2-H19 DMRs was not found to be significantly different among two groups of bulls. The sixth CTCF binding site (CCCTC) identified in H19 DMR, however, had a significant methylation difference between the high- and low-fertility bulls. It was concluded that alteration of the methylation levels at IGF2-H19 DMRs might not be responsible for the fertility difference of crossbred bulls, although the role played by the specific CTCF binding sites at this locus, which could influence IGF2 expression during spermatogenesis and early embryonic development, deserves further attention.

  14. p16 methylation does not affect protein expression in cervical carcinogenesis.

    Science.gov (United States)

    Nehls, Kristina; Vinokurova, Svetlana; Schmidt, Dietmar; Kommoss, Friedrich; Reuschenbach, Miriam; Kisseljov, Fjodor; Einenkel, Jens; von Knebel Doeberitz, Magnus; Wentzensen, Nicolas

    2008-11-01

    Previous studies have reported a frequency range of 19-61% for p16 methylation in cervical cancers. However, p16 is strongly expressed in over 90% of cervical cancers and pre-cancers, due to interactions of HPV oncogenes with p53 and pRb. In order to clarify these controversial findings, we developed a new bisulphite sequencing protocol to determine the methylation status of p16. DNA extracted from 17 cell lines and 94 microdissected clinical samples was subjected to methylation analysis. p16 expression was confirmed in Western blot and immunohistochemistry. Complete methylation of p16 was found in none of the dysplastic lesions, but in 26% of the cervical carcinomas. However, immunohistochemistry showed strong p16 expression in all cancers. These findings indicate that p16 methylation does not implicate loss of p16 expression in HPV-induced tumours. In cervical cancer, methylation of p16 does not seem to be an underlying pathogenic mechanism, but may be a result of increasing genetic and epigenetic instability.

  15. A histochemical comparison of methyl green-pyronin, and hematoxylin and eosin for detecting apoptotic cells in oral squamous cell carcinoma, oral leukoplakia, oral submucous fibrosis and normal oral mucosa.

    Science.gov (United States)

    Sumedha, S; Kotrashetti, V S; Somannavar, P; Nayak, R; Babji, D

    2015-05-01

    Analysis of apoptotic cells in oral pathological states could be useful for determining the rates of tissue turnover, which would help determine prognosis. The use of histochemical stains such as hematoxylin and eosin (H & E) and methyl green-pyronin (MGP) can provide a simple and cost-effective method for detecting apoptotic cells. We compared the efficacy of MGP and H & E for detecting apoptotic cells in oral squamous cell carcinoma (OSCC), oral leukoplakia (OL), oral submucous fibrosis (OSMF) and normal oral mucosa (NOM). Ten cases each of OSCC, OSMF, OL and NOM were retrieved from the archives and two serial sections were stained, one with H & E and the other with MGP. Apoptotic cells were identified at 100 x magnification and the apoptotic index was calculated. Apoptotic cells were distinguished more readily in MGP stained sections than in those stained with H & E. Also, the apoptotic cell count was greater in OSCC compared to OL, OSMF and NOM. We concluded that MGP staining can be used as a routine, cost-effective method for detecting apoptotic cells.

  16. The Hepatitis Viral Status in Patients With Hepatocellular Carcinoma: a Study of 3843 Patients From Taiwan Liver Cancer Network

    Science.gov (United States)

    Chang, Il-Chi; Huang, Shiu-Feng; Chen, Pei-Jer; Chen, Chi-Ling; Chen, Chao-Long; Wu, Cheng-Chung; Tsai, Cheng-Chung; Lee, Po-Huang; Chen, Miin-Fu; Lee, Chuan-Mo; Yu, Hsien-Chung; Lo, Gin-Ho; Yeh, Chau-Ting; Hong, Chih-Chen; Eng, Hock-Liew; Wang, John; Tseng, Hui-Hwa; Hsiao, Cheng-Hsiang; Wu, Hong-Dar Isaac; Yen, Tseng-Chang; Liaw, Yun-Fan

    2016-01-01

    Abstract Hepatocellular carcinoma (HCC) is the leading cancer death in Taiwan. Chronic viral hepatitis infections have long been considered as the most important risk factors for HCC in Taiwan. The previously published reports were either carried out by individual investigators with small patient numbers or by large endemic studies with limited viral marker data. Through collaboration with 5 medical centers across Taiwan, Taiwan liver cancer network (TLCN) was established in 2005. All participating centers followed a standard protocol to recruit liver cancer patients along with their biosamples and clinical data. In addition, detailed viral marker analysis for hepatitis B virus (HBV) and hepatitis C virus (HCV) were also performed. This study included 3843 HCC patients with available blood samples in TLCN (recruited from November 2005 to April 2011). There were 2153 (56.02%) patients associated with HBV (HBV group); 969 (25.21%) with HCV (HCV group); 310 (8.07%) with both HBV and HCV (HBV+HCV group); and 411 (10.69%) were negative for both HBV and HCV (non-B non-C group). Two hundred two of the 2463 HBV patients (8.20%) were HBsAg(-), but HBV DNA (+). The age, gender, cirrhosis, viral titers, and viral genotypes were all significantly different between the above 4 groups of patients. The median age of the HBV group was the youngest, and the cirrhotic rate was lowest in the non-B non-C group (only 25%). This is the largest detailed viral hepatitis marker study for HCC patients in the English literatures. Our study provided novel data on the interaction of HBV and HCV in the HCC patients and also confirmed that the HCC database of TLCN is highly representative for Taiwan and an important resource for HCC research. PMID:27082566

  17. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

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    F.E. Rosa

    2013-03-01

    Full Text Available Human epidermal growth factor receptor 2 (HER2 has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC. HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH in moderate immunoexpression (IHC 2+ cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH, which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

  18. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    Directory of Open Access Journals (Sweden)

    F.E. Rosa

    Full Text Available Human epidermal growth factor receptor 2 (HER2 has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC. HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH in moderate immunoexpression (IHC 2+ cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH, which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

  19. Chromogenic in situ hybridization compared with other approaches to evaluate HER2/neu status in breast carcinomas

    Energy Technology Data Exchange (ETDEWEB)

    Rosa, F.E.; Santos, R.M. [Departamento de Patologia, Hospital das Clínicas, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil); Rogatto, S.R. [Departamento de Urologia, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil); Hospital A.C. Camargo, CIPE, São Paulo, SP (Brazil); Domingues, M.A.C. [Departamento de Patologia, Faculdade de Medicina, Universidade Estadual Paulista, Botucatu, SP (Brazil)

    2013-03-19

    Human epidermal growth factor receptor 2 (HER2) has been evaluated in breast cancer patients to identify those most likely to benefit from herceptin-targeted therapy. HER2 amplification, detected in 20-30% of invasive breast tumors, is associated with reduced survival and metastasis. The most frequently used technique for evaluating HER2 protein status as a routine procedure is immunohistochemistry (IHC). HER2 copy number alterations have also been evaluated by fluorescence in situ hybridization (FISH) in moderate immunoexpression (IHC 2+) cases. An alternative procedure to evaluate gene amplification is chromogenic in situ hybridization (CISH), which has some advantages over FISH, including the correlation between HER2 status and morphological features. Other methodologies have also been used, such as silver-enhanced in situ hybridization (SISH) and quantitative real-time RT-PCR, to determine the number of HER2 gene copies and expression, respectively. Here we will present a short and comprehensive review of the current advances concerning HER2 evaluation in human breast cancer.

  20. Effects of 5-aza-CdR on methylation and expression of PDCD4 gene in hepatocellular carcinoma cell line SMMC7721%5-aza-CdR对肝癌SMMC7721细胞株PDCD4基因甲基化及表达的影响

    Institute of Scientific and Technical Information of China (English)

    王家祥; 戴兵; 刘秋亮; 郭广成; 王佳辰

    2009-01-01

    Objective To investigate the effects of 5-aza-CdR on expression of programmed cell death factor 4 (PDCD4) in SMMC7721and explore its possible mechanism. Methods Hepatocellular carcinoma cell line SMMC7721 was cultivated with 5-aza-CdR inhibiting the expression of DNMT3b. Western blotting was used to determine the changes of DNMT3b and PDCD4 proteins and the methylation level of PDCD4's promoter was tested by methylation specific polymerase chain reaction(MSP). Results Using the 5-aza-CdR on concentration of 1×10-6mol/L and 5×10-6mol/L to cultivate SMMC7721, the expression of DNMT3b protein was decreased while the PDCD4 protein was increased. The methylation level of PDCD4's promoter was high in SMMC7721. However, after the treatment of 5-aza-CdR on concentration of 5×10-6mol/L, the promoter of PDCD4 had demethylation. Conclusion The expression of DNMT3b can be inhibited by 5-aza-CdR and DNMT3b may control the expression of PDCD4 in SMMC7721 by influencing the methylation status of its promoter.%目的 研究5-氮杂-2'-脱氧胞苷(5-aza-CdR)在肝癌细胞株SMMC7721中对程序性细胞死亡因子4(PDCD4)表达的影响及可能机制.方法 培养肝癌细胞株SMMC7721,用5-aza-CdR处理肝癌细胞株SMMC7721,Western-blotting检测DNMT3b、PDCD4蛋白在处理前后的变化,甲基化特异性PCR即MSP检测PDCD4基因启动子区域甲基化水平.结果 1×10~(-6)mol/L、5×10~(-6)mol/L的5-aza-CdR作用于SMMC7721细胞后,DNMT3b表达水平降低,而PDCD4表达水平升高,且浓度之间有差异;MSP示药物处理前PDCD4启动子区域处于高甲基化水平,在用5×10~(-6)mol/L药物处理后,其启动子发生了去甲基化.结论 5-aza-CdR可以抑制DNMT3b在SMMC7721中的表达,且可能通过改变抑癌基因PDCD4启动子甲基化状态影响PDCD4表达.

  1. 微小RNA-375在食管鳞状细胞癌组织中的表达及其甲基化调控%Expression of microRNA-375 in esophageal squamous cell carcinoma and its methylation regulation

    Institute of Scientific and Technical Information of China (English)

    董新伟; 银瑞; 卢万里

    2014-01-01

    目的 观察微小RNA(miRNA)-375在食管鳞状细胞癌(ESCC)中的表达,探讨其启动子区的甲基化水平与ESCC发生的关系.方法 对62例ESCC组织标本和正常食管组织(距癌肿边缘>3 cm)进行实时荧光定量聚合酶链反应(FQ-PCR)检测,使用甲基化特异性聚合酶链反应(MSP)检测样本中miRNA-375上游启动子区的甲基化.统计学分析miRNA-375的表达与临床因素的相关性.结果 62例ESCC组织标本中有53例miRNA-375E呈明显低表达(85%).MSP结果显示,ESCC组织的甲基化水平明显高于正常食管组织,差异有统计学意义(P<0.05).与临床因素的相关分析显示,miRNA-375在ESCC组织中低表达,与ESCC的TNM分期呈负相关(P<0.05).结论 ESCC组织中存在miRNA-375基因异常低表达及其启动子区的高甲基化,可能在ESCC发生发展中发挥重要作用.%Objective To investigate the expression of microRNA (miRNA)-375 and its genomic promoter DNA methylation in esophageal squamous cell carcinoma (ESCC).Methods The expression of miRNA-375 in ESCC and the matched normal esophageal tissues (from the tumor edge > 3 cm) was detected by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR).DNA methylation status was examined by methylation specific polymerease chain reaction (MSP).The association between miRNA-375 expression and clinical factors were analyzed statistically.Results As compared with normal controls,miRNA-375 was significantly decreased in ESCC.The methylation levels in ESCC tissues were significantly higher than in the matched normal esophageal tissues (P < 0.05).The low expression of miRNA-375 was significandy associated with ESCC TNM stage (P < 0.05).Conclusion The lower expression of miRNA-375 and aberrant hypermethylation of the CpG island promoters in ESCC may play a crucial role in the carcinogenesis of ESCC.

  2. Associations between methylation of paternally expressed gene 3 (PEG3, cervical intraepithelial neoplasia and invasive cervical cancer.

    Directory of Open Access Journals (Sweden)

    Monica D Nye

    Full Text Available Cytology-based screening for invasive cervical cancer (ICC lacks sensitivity and specificity to discriminate between cervical intraepithelial neoplasia (CIN likely to persist or progress from cases likely to resolve. Genome-wide approaches have been used to identify DNA methylation marks associated with CIN persistence or progression. However, associations between DNA methylation marks and CIN or ICC remain weak and inconsistent. Between 2008-2009, we conducted a hospital-based, case-control study among 213 Tanzania women with CIN 1/2/3 or ICC. We collected questionnaire data, biopsies, peripheral blood, cervical scrapes, Human papillomavirus (HPV and HIV-1 infection status. We assessed PEG3 methylation status by bisulfite pyrosequencing. Multinomial logistic regression was used to estimate odds ratios (OR and confidence intervals (CI 95% for associations between PEG3 methylation status and CIN or ICC. After adjusting for age, gravidity, hormonal contraceptive use and HPV infection, a 5% increase in PEG3 DNA methylation was associated with increased risk for ICC (OR = 1.6; 95% CI 1.2-2.1. HPV infection was associated with a higher risk of CIN1-3 (OR = 15.7; 95% CI 5.7-48.6 and ICC (OR = 29.5, 95% CI 6.3-38.4. Infection with high risk HPV was correlated with mean PEG3 differentially methylated regions (DMRs methylation (r = 0.34 p<0.0001, while the correlation with low risk HPV infection was weaker (r = 0.16 p = 0.047. Although small sample size limits inference, these data support that PEG3 methylation status has potential as a molecular target for inclusion in CIN screening to improve prediction of progression. Impact statement: We present the first evidence that aberrant methylation of the PEG3 DMR is an important co-factor in the development of Invasive cervical carcinoma (ICC, especially among women infected with high risk HPV. Our results show that a five percent increase in DNA methylation of PEG3 is associated with

  3. Associations between Methylation of Paternally Expressed Gene 3 (PEG3), Cervical Intraepithelial Neoplasia and Invasive Cervical Cancer

    Science.gov (United States)

    Nye, Monica D.; Hoyo, Cathrine; Huang, Zhiqing; Vidal, Adriana C.; Wang, Frances; Overcash, Francine; Smith, Jennifer S.; Vasquez, Brandi; Hernandez, Brenda; Swai, Britta; Oneko, Olola; Mlay, Pendo; Obure, Joseph; Gammon, Marilie D.; Bartlett, John A.; Murphy, Susan K.

    2013-01-01

    Cytology-based screening for invasive cervical cancer (ICC) lacks sensitivity and specificity to discriminate between cervical intraepithelial neoplasia (CIN) likely to persist or progress from cases likely to resolve. Genome-wide approaches have been used to identify DNA methylation marks associated with CIN persistence or progression. However, associations between DNA methylation marks and CIN or ICC remain weak and inconsistent. Between 2008–2009, we conducted a hospital-based, case-control study among 213 Tanzania women with CIN 1/2/3 or ICC. We collected questionnaire data, biopsies, peripheral blood, cervical scrapes, Human papillomavirus (HPV) and HIV-1 infection status. We assessed PEG3 methylation status by bisulfite pyrosequencing. Multinomial logistic regression was used to estimate odds ratios (OR) and confidence intervals (CI 95%) for associations between PEG3 methylation status and CIN or ICC. After adjusting for age, gravidity, hormonal contraceptive use and HPV infection, a 5% increase in PEG3 DNA methylation was associated with increased risk for ICC (OR = 1.6; 95% CI 1.2–2.1). HPV infection was associated with a higher risk of CIN1-3 (OR = 15.7; 95% CI 5.7–48.6) and ICC (OR = 29.5, 95% CI 6.3–38.4). Infection with high risk HPV was correlated with mean PEG3 differentially methylated regions (DMRs) methylation (r = 0.34 p<0.0001), while the correlation with low risk HPV infection was weaker (r = 0.16 p = 0.047). Although small sample size limits inference, these data support that PEG3 methylation status has potential as a molecular target for inclusion in CIN screening to improve prediction of progression. Impact statement We present the first evidence that aberrant methylation of the PEG3 DMR is an important co-factor in the development of Invasive cervical carcinoma (ICC), especially among women infected with high risk HPV. Our results show that a five percent increase in DNA methylation of PEG3 is associated with

  4. Relationship between The Kiss-1 Gene Promoter Methylation and Kiss-1 Gene Expression of Colorectal Carcinoma%结直肠癌中Kiss-1基因启动子甲基化与Kiss-1基因表达的关系

    Institute of Scientific and Technical Information of China (English)

    林素勇; 陈志华; 陈绍勤; 戴起宝

    2013-01-01

    目的:研究结直肠癌中Kiss-1基因启动子甲基化状态对Kiss-1基因表达的影响。方法:应用甲基化特异性PCR (MSP)方法检测73例结直肠癌、正常结直肠组织和人结直肠癌细胞HCT116、SW480、W1116、LoVo中Kiss-1基因启动子甲基化状态,应用realtime-PCR、Western-blot技术检测相应组织和细胞中Kiss-1基因mRNA和蛋白质(Metastine)的表达量。结果:结直肠癌中Kiss-1基因甲基化阳性率(82.19%)高于正常组织(6.31%)(PSW480>SW1116>HCT116,差异有统计学意义(p<0.05)。结论:结直肠癌中Kiss-1基因启动子甲基化可能引起Kiss-1基因表达下调。%Objective:To research the effect of the Kiss-1 gene promoter methylation on Kiss-1 gene expression in colorectal carci-noma. Methods:Kiss-1 gene promotor methylation and its expression were detected respectively by methylation-specific PCR, real-time PCR and western-blot in 73 cases of colorectal carcinoma and each normal tissue. Results:The positive expression rate of Kiss-1 gene methylation in the carcinoma tissue was higher than the rate in normal tissue (82.19%vs 6.31%, P <0.05). In the cancer tissue, the Kiss-1 gene mRNA and metastine expression of Kiss-1 methylation-positive group was significant lower than that in negative group (P<0.05). Conclusions:Kiss-1 gene promoter hyper-methylation may induce to the Kiss-1 gene expression decreased in colorectal carci-noma.

  5. Effect of kaempferol on lipid peroxidation and antioxidant status in 1,2-dimethyl hydrazine induced colorectal carcinoma in rats.

    Science.gov (United States)

    Nirmala, Parthasarathy; Ramanathan, Manickam

    2011-03-01

    Colorectal cancer, a common cause of cancer related deaths in both sexes in western population is often due to persistent oxidative stress leading to DNA damage. Antioxidants scavenge free radicals and inhibit neoplastic process. Kaempferol, a flavonol widely distributed in tea, broccoli, grape fruit, brussels sprouts and apple, is claimed to have chemopreventive action in colon cancer. The aim of our study was to evaluate the effect of kaempferol on tissue lipid peroxidation and antioxidant status in 1,2-dimethyl hydrazine induced colorectal cancer in male Wistar rats and to compare its efficacy with irinotecan. Experimental colon cancer induced by 1,2-dimethyl hydrazine in rats mimic human colon cancer and therefore is an ideal model for chemoprevention studies. The rats were divided into six groups. Group 1 served as control. Group 2 received 1,2-dimethyl hydrazine (20 mg/kg body weight) subcutaneously once a week for four weeks. Group 3 received irinotecan (100 mg/kg body weight) intravenously once a week for four weeks with 1,2-dimethyl hydrazine. Groups 4 to 6 were given a daily oral dose of 50, 100, 200 mg/kg body weight of kaempferol with 1,2-dimethyl hydrazine. The total study period was 16 weeks. Kaempferol supplementation lowered 1,2-dimethyl hydrazine induced erythrocyte lysate and liver thiobarbituric acid reactive substances level and rejuvenated anti oxidant enzymes catalase, super oxide dismutase and glutathione peroxidase. The recovery of enzyme status was maximum at the dose of 200 mg/kg body weight and was comparable to irinotecan. Our study reveals that kaempferol could be safely used as a chemopreventive agent in colorectal cancer.

  6. High-level microsatellite instability in appendiceal carcinomas.

    Science.gov (United States)

    Taggart, Melissa W; Galbincea, John; Mansfield, Paul F; Fournier, Keith F; Royal, Richard E; Overman, Michael J; Rashid, Asif; Abraham, Susan C

    2013-08-01

    High-level microsatellite instability (MSI-high) is found in approximately 15% of all colorectal adenocarcinomas (CRCs) and in at least 20% of right-sided cancers. It is most commonly due to somatic hypermethylation of the MLH1 gene promoter region, with familial cases (Lynch syndrome) representing only 2% to 3% of CRCs overall. In contrast to CRC, MSI-high in appendiceal adenocarcinomas is rare. Only 4 MSI-high appendiceal carcinomas and 1 MSI-high appendiceal serrated adenoma have been previously reported, and the prevalence of MSI in the appendix is unknown. We identified 108 appendiceal carcinomas from MD Anderson Cancer Center in which MSI status had been assessed by immunohistochemistry for the DNA mismatch-repair proteins MLH1, MSH2, MSH6, and PMS2 (n=83), polymerase chain reaction (n=7), or both (n=18). Three cases (2.8%) were MSI-high, and 1 was MSI-low. The 3 MSI-high cases included: (1) a poorly differentiated nonmucinous adenocarcinoma with loss of MLH1/PMS2 expression, lack of MLH1 promoter methylation, and lack of BRAF gene mutation, but no detected germline mutation in MLH1 from a 39-year-old man; (2) an undifferentiated carcinoma with loss of MSH2/MSH6, but no detected germline mutation in MSH2 or TACSTD1, from a 59-year-old woman; and (3) a moderately differentiated mucinous adenocarcinoma arising in a villous adenoma with loss of MSH2/MSH6 expression, in a 38-year-old man with a strong family history of CRC who declined germline testing. When the overall group of appendiceal carcinomas was classified according to histologic features and precursor lesions, the frequencies of MSI-high were: 3 of 108 (2.8%) invasive carcinomas, 3 of 96 (3.1%) invasive carcinomas that did not arise from a background of goblet cell carcinoid tumors, and 0 of 12 (0%) signet ring and mucinous carcinomas arising in goblet cell carcinoid tumors. These findings, in conjunction with the previously reported MSI-high appendiceal carcinomas, highlight the low prevalence of MSI

  7. Relationship of methylation status in promoter of APC gene and environmental factors with prostate cancer%APC基因启动子区甲基化及环境因素与前列腺癌的关系

    Institute of Scientific and Technical Information of China (English)

    邱镇; 张连升; 梁永; 崔飞伦

    2013-01-01

    目的 探讨腺瘤样结肠息肉易感基因(APC)启动子区CpG甲基化及环境危险因素及相互作用与中国人群中前列腺癌(PCa)发病之间的关系.方法 应用亚硫酸盐修饰后测序法检测60例PCa及40例良性前列腺增生(BPH)组织中APC基因启动子区CpG甲基化情况.分析APC基因甲基化及环境危险因素与PCa患病之间的关系.结果 APC基因在PCa和BPH组织中CG位点甲基化率分别为48.84%和1.19%;基因甲基化率与前列腺特异性抗原(PSA)、Gleason评分、病理分期和PCa危险分期之间关系密切;饮茶者患PCa的危险性是不饮茶者的0.29倍.结论 APC基因启动子区甲基化与PCa发生及发展有关;其甲基化率的变化与PCa的危险分期关系密切.检测前列腺组织中相关基因甲基化状态,有望成为诊断早期PCa的一种方法.%Objective To investigate the relationship of methylation status in promoter of adenomatosis polyposis coli(APC) gene and environmental factors with prostate cancer (PCa). Methods With bisulphite sequencing PCR,the methylation of APC gene was detected in PCa tissues of 60 PCa patients and hyperplastic tissues of 40 patients with benign prostate hyperplasia(BPH). The methylation status and environmental risk factors for PCa were analyzed. Results The methylation rates of CG sites in promoter of APC gene in PCa and BPH tissues were 48.84% and 1. 19%, respectively. The methylation rate of APC gene was closely associated with prostate specific antigen (PSA),Gleason score,pathologic stage and classification of the risks. A decreased risk for PCa was observed in Chinese population having a tea drinking habit. Conclusion The methylation of promoter in APC genes is associated with the occurrence and progress of PCa The change of methylation rate of APC genes is closely related to the classification of PCa risks. Detection of the methylation status of APC genes may hopefully become an indicator for early diagnosis of PCa.

  8. Genome-wide screening identifies Plasmodium chabaudi-induced modifications of DNA methylation status of Tlr1 and Tlr6 gene promoters in liver, but not spleen, of female C57BL/6 mice.

    Science.gov (United States)

    Al-Quraishy, Saleh; Dkhil, Mohamed A; Abdel-Baki, Abdel Azeem S; Delic, Denis; Santourlidis, Simeon; Wunderlich, Frank

    2013-11-01

    Epigenetic reprogramming of host genes via DNA methylation is increasingly recognized as critical for the outcome of diverse infectious diseases, but information for malaria is not yet available. Here, we investigate the effect of blood-stage malaria of Plasmodium chabaudi on the DNA methylation status of host gene promoters on a genome-wide scale using methylated DNA immunoprecipitation and Nimblegen microarrays containing 2,000 bp oligonucleotide features that were split into -1,500 to -500 bp Ups promoters and -500 to +500 bp Cor promoters, relative to the transcription site, for evaluation of differential DNA methylation. Gene expression was analyzed by Agilent and Affymetrix microarray technology. Challenging of female C57BL/6 mice with 10(6) P. chabaudi-infected erythrocytes resulted in a self-healing outcome of infections with peak parasitemia on day 8 p.i. These infections induced organ-specific modifications of DNA methylation of gene promoters. Among the 17,354 features on Nimblegen arrays, only seven gene promoters were identified to be hypermethylated in the spleen, whereas the liver exhibited 109 hyper- and 67 hypomethylated promoters at peak parasitemia in comparison with non-infected mice. Among the identified genes with differentially methylated Cor-promoters, only the 7 genes Pigr, Ncf1, Klkb1, Emr1, Ndufb11, and Tlr6 in the liver and Apol6 in the spleen were detected to have significantly changed their expression. Remarkably, the Cor promoter of the toll-like receptor Tlr6 became hypomethylated and Tlr6 expression increased by 3.4-fold during infection. Concomitantly, the Ups promoter of the Tlr1 was hypermethylated, but Tlr1 expression also increased by 11.3-fold. TLR6 and TLR1 are known as auxillary receptors to form heterodimers with TLR2 in plasma membranes of macrophages, which recognize different pathogen-associated molecular patterns (PAMPs), as, e.g., intact 3-acyl and sn-2-lyso-acyl glycosylphosphatidylinositols of P. falciparum

  9. 沉默DNMT1和DNMT3b基因对胰腺癌BxPC-3细胞p16,RASSF1A基因启动子甲基化的影响%Effect of gene silencing of DNMT1 and DNMT3b on the methylation in the promoter region of p 16 and RASSF1A gene in pancreatic carcinoma BxPC-3 cells

    Institute of Scientific and Technical Information of China (English)

    肖卫东; 李勇; 邹叶青; 李学明; 蔡军; 曾林山; 胡伟

    2012-01-01

    目的:探讨沉默DNMT1和DNMT3b基因对胰腺癌BxPC-3细胞p16,RASSF1A基因启动子甲基化的影响.方法:将胰腺癌BxPC-3细胞分为5组:DNMT1干扰组(转染DNMT1-siRNA),DNMT3b干扰组(转染DNMT3b-siRNA),双重干扰组(转染DNMT1+DNMT3b-siRNA),阴性对照组(转染阴性siRNA)和空白对照组(转染脂质体).转染48 h后,应用荧光定最PCR法和Western blot法分别检测细胞中DNMT1和DNMT3b的mRNA及蛋白的表达水平;甲基化特异性PCR法检测p16和RASSF1A基因启动子甲基化.结果:与空白对照组和阴性对照组比较,各干扰组目的基因的mRNA及蛋白表达量均明显降低(均P<0.01).空白对照组与阴性对照组p16和RASSF1A基因甲基化阳性;DNMT1干扰组和双重干扰组p16基因甲基化阴性,RASSF1A基因部分甲基化;DNMT3b干扰组p16基因部分甲基化,RASSF1A基因甲基化阳性.结论:DNMT1单干扰对胰腺癌BxPC-3细胞p16和RASSF1A基因的去甲基化作用优于DNMT3b单干扰,DNMT1和DNMT3b双重干扰无明显的协同效应;提示DNMT1是胰腺癌去甲基化治疗的一个有效靶点.%Objective: To investigate the effect of gene silencing of DNMT1 and DNMT3b on methylation in the promoter region of pl6 and RASSF1A gene in pancreatic carcinoma BxPC-3 cells.Methods: The pancreatic carcinoma BxPC-3 cells were divided into five groups, which included the DNMT1 interference group (transfected with DNMTl-siRNA), DNMT3b interference group (transfected with DNMT3b-siRNA), double interference group (transfected with DNMTl+DNMT3b-siRNA), negative control group (transfected negative-siRNA) and blank control group (transfected with lipofectamine). Forty-eight hours after transfection, the mRNA and protein expression of DNMT1 and DNMT3b were analyzed by real-time RT-PCR and Western blot, and the methylation status in the promoter region of pl6 and RASSF1A gene was detected with methylation-specific PCR (MSP).Results: Compared with the negative or blank control group, the m

  10. The effects of depression and use of antidepressive medicines during pregnancy on the methylation status of the IGF2 imprinted control regions in the offspring

    Directory of Open Access Journals (Sweden)

    Soubry A

    2011-10-01

    Full Text Available Abstract In utero exposures to environmental factors may result in persistent epigenetic modifications affecting normal development and susceptibility to chronic diseases in later life. We explored the relationship between exposure of the growing fetus to maternal depression or antidepressants and DNA methylation at two differentially methylated regions (DMRs of the imprinted Insulin-like Growth Factor 2 (IGF2 gene. Aberrant DNA methylation at the IGF2 and neighboring H19 DMRs has been associated with deregulated IGF2 expression, childhood cancers and several chronic diseases during adulthood. Our study population is comprised of pregnant mothers and their newborns (n = 436, as part of the Newborn Epigenetics Study (NEST. A standardized questionnaire was completed and medical record data were abstracted to ascertain maternal depression and antidepressive drug use. DMR methylation levels in umbilical cord blood leukocytes were quantified using pyrosequencing. From the 436 newborns, laboratory data were obtained for 356 individuals at the IGF2 DMRs, and for 411 individuals at the H19 DMRs; about half of each group was African American or Caucasian. While overall no association between depression and methylation profiles was found, we observed a significant hypermethylation of the H19 DMRs in newborns of African American (n = 177 but not Caucasian (n = 168 mothers who reported the use of antidepressive drugs during pregnancy (β = +6.89, p = 0.01. Of note, our data reveal a race-independent association between smoking during pregnancy and methylation at the IGF2 DMR (+3.05%, p = 0.01. In conclusion, our findings suggest a race-dependent response related to maternal use of antidepressants at one of the IGF2 DMRs in the offspring.

  11. The Zinc Concentration in the Diet and the Length of the Feeding Period Affect the Methylation Status of the ZIP4 Zinc Transporter Gene in Piglets.

    Directory of Open Access Journals (Sweden)

    Diana Karweina

    Full Text Available High doses of zinc oxide are commonly used in weaned pig diets to improve performance and health. Recent reports show that this may also lead to an imbalanced zinc homeostasis in the animal. For a better understanding of the regulatory mechanisms of different zinc intakes, we performed a feeding experiment to assess potential epigenetic regulation of the ZIP4 gene expression via DNA methylation in the small intestine of piglets. Fifty-four piglets were fed diets with 57 (LZn, 164 (NZn or 2,425 (HZn mg Zn/kg feed for one or four weeks. The ZIP4 expression data provided significant evidence for counter-regulation of zinc absorption with higher dietary zinc concentrations. The CpG +735 in the second exon had a 56% higher methylation in the HZn group compared to the others after one week of feeding (8.0·10-4 < p < 0.035; the methylation of this CpG was strongly negatively associated with the expression of the long ZIP4 transcripts (p < 0.007. In the LZn and NZn diets, the expression of the long ZIP4 transcripts were lower after four vs. one week of feeding (2.9·10-4 < p < 0.017. The strongest switch leading to high DNA methylation in nearly all analysed regions was dependent on feeding duration or age in all diet groups (3.7·10-10 < p < 0.099. The data suggest that DNA methylation serves as a fine-tuning mechanism of ZIP4 gene regulation to maintain zinc homeostasis. Methylation of the ZIP4 gene may play a minor role in the response to very high dietary zinc concentration, but may affect binding of alternate zinc-responsive transcription factors.

  12. The Zinc Concentration in the Diet and the Length of the Feeding Period Affect the Methylation Status of the ZIP4 Zinc Transporter Gene in Piglets.

    Science.gov (United States)

    Karweina, Diana; Kreuzer-Redmer, Susanne; Müller, Uwe; Franken, Tobias; Pieper, Robert; Baron, Udo; Olek, Sven; Zentek, Jürgen; Brockmann, Gudrun A

    2015-01-01

    High doses of zinc oxide are commonly used in weaned pig diets to improve performance and health. Recent reports show that this may also lead to an imbalanced zinc homeostasis in the animal. For a better understanding of the regulatory mechanisms of different zinc intakes, we performed a feeding experiment to assess potential epigenetic regulation of the ZIP4 gene expression via DNA methylation in the small intestine of piglets. Fifty-four piglets were fed diets with 57 (LZn), 164 (NZn) or 2,425 (HZn) mg Zn/kg feed for one or four weeks. The ZIP4 expression data provided significant evidence for counter-regulation of zinc absorption with higher dietary zinc concentrations. The CpG +735 in the second exon had a 56% higher methylation in the HZn group compared to the others after one week of feeding (8.0·10-4 < p < 0.035); the methylation of this CpG was strongly negatively associated with the expression of the long ZIP4 transcripts (p < 0.007). In the LZn and NZn diets, the expression of the long ZIP4 transcripts were lower after four vs. one week of feeding (2.9·10-4 < p < 0.017). The strongest switch leading to high DNA methylation in nearly all analysed regions was dependent on feeding duration or age in all diet groups (3.7·10-10 < p < 0.099). The data suggest that DNA methylation serves as a fine-tuning mechanism of ZIP4 gene regulation to maintain zinc homeostasis. Methylation of the ZIP4 gene may play a minor role in the response to very high dietary zinc concentration, but may affect binding of alternate zinc-responsive transcription factors.

  13. A CpG island hypermethylation profile of primary colorectal carcinomas and colon cancer cell lines

    Directory of Open Access Journals (Sweden)

    Rognum Torleiv O

    2004-10-01

    Full Text Available Abstract Background Tumor cell lines are commonly used as experimental tools in cancer research, but their relevance for the in vivo situation is debated. In a series of 11 microsatellite stable (MSS and 9 microsatellite unstable (MSI colon cancer cell lines and primary colon carcinomas (25 MSS and 28 MSI with known ploidy stem line and APC, KRAS, and TP53 mutation status, we analyzed the promoter methylation of the following genes: hMLH1, MGMT, p16INK4a (CDKN2A α-transcript, p14ARF (CDKN2A β-transcript, APC, and E-cadherin (CDH1. We compared the DNA methylation profiles of the cell lines with those of the primary tumors. Finally, we examined if the epigenetic changes were associated with known genetic markers and/or clinicopathological variables. Results The cell lines and primary tumors generally showed similar overall distribution and frequencies of gene methylation. Among the cell lines, 15%, 50%, 75%, 65%, 20% and 15% showed promoter methylation for hMLH1, MGMT, p16INK4a, p14ARF, APC, and E-cadherin, respectively, whereas 21%, 40%, 32%, 38%, 32%, and 40% of the primary tumors were methylated for the same genes. hMLH1 and p14ARF were significantly more often methylated in MSI than in MSS primary tumors, whereas the remaining four genes showed similar methylation frequencies in the two groups. Methylation of p14ARF, which indirectly inactivates TP53, was seen more frequently in tumors with normal TP53 than in mutated samples, but the difference was not statistically significant. Methylation of p14ARF and p16INK4a was often present in the same primary tumors, but association to diploidy, MSI, right-sided location and female gender was only significant for p14ARF. E-cadherin was methylated in 14/34 tumors with altered APC further stimulating WNT signaling. Conclusions The present study shows that colon cancer cell lines are in general relevant in vitro models, comparable with the in vivo situation, as the cell lines display many of the same

  14. Real-life practice study of the clinical outcome and cost-effectiveness of photodynamic therapy using methyl aminolevulinate (MAL-PDT) in the management of actinic keratosis and basal cell carcinoma.

    Science.gov (United States)

    Annemans, Lieven; Caekelbergh, Karin; Roelandts, Rik; Boonen, Hugo; Leys, Christoph; Nikkels, Arjen F; van Den Haute, V; van Quickenborne, L; Verhaeghe, Evelien; Leroy, Bernard

    2008-01-01

    Clinical trials have shown that photodynamic therapy using methyl aminolevulinate (MAL-PDT) is an effective treatment for actinic keratosis (AK), and nodular and superficial basal cell carcinoma (nBCC and sBCC) unsuitable for other available therapies. Economic evaluation models have shown that it is a cost effective intervention as well. The objectives of this prospective, observational, one arm study were (i) to verify in a real-life practice study the results obtained in previous clinical trials with MAL-PDT in the treatment of AK, nBCC and sBCC; (ii) to calculate the real-life cost of treatment and validate predictions from an economic evaluation model. Patients with AK and/or BCC were selected according to Belgian reimbursement criteria for treatment with MAL-PDT. Clinical response, cosmetic outcome and tolerability were assessed. MAL-PDT cost was calculated and compared to published model cost data. Data were collected from 247 patients (117 AK, 130 BCC). A complete clinical response was obtained for 83% of AK (85/102) and BCC (97/116) patients. A good or excellent cosmetic outcome was obtained for 95% of AK patients and 93% of BCC patients. Tolerability was good: only 2 patients withdrew for adverse events. Clinical results were similar to previous studies. Total cost of care per patient was euro 381 for AK, euro 318 for nBCC, and euro 298 for sBCC. Total cost per lesion was euro 58 for AK (identical to model prediction), euro 316 for nBCC and euro 178 for sBCC (both within 20% of model prediction). The clinical results of MAL-PDT in this real-life practice study confirm those demonstrated in previous clinical trials. Costs calculated from this study confirm predicted cost-effectiveness in the original model for MAL-PDT in the management of AK and BCC.

  15. The combination of IDH1 mutations and MGMT methylation status predicts survival in glioblastoma better than either IDH1 or MGMT alone

    NARCIS (Netherlands)

    Molenaar, R.J.; Verbaan, D.; Lamba, S.; Zanon, C.; Jeuken, J.W.M.; Boots-Sprenger, S.H.E.; Wesseling, P.; Hulsebos, T.J.M.; Troost, D.; Tilborg, A.A. Van; Leenstra, S.; Vandertop, W.P.; Bardelli, A.; Noorden, C.J.F. van; Bleeker, F.E.

    2014-01-01

    BACKGROUND: Genetic and epigenetic profiling of glioblastomas has provided a comprehensive list of altered cancer genes of which only O(6)-methylguanine-methyltransferase (MGMT) methylation is used thus far as a predictive marker in a clinical setting. We investigated the prognostic significance of

  16. The combination of IDH1 mutations and MGMT methylation status predicts survival in glioblastoma better than either IDH1 or MGMT alone

    NARCIS (Netherlands)

    R.J. Molenaar (Remco J.); D. Verbaan (Dagmar); S. Lamba (Simona); C. Zanon (Carlo); J. Jeuken (Judith); S.H.E. Boots-Sprenger (Sandra H. E.); P. Wesseling (Pieter); T. Hulsebos (Theo); D. Troost (Dirk); A.A.G. van Tilborg (Angela); S. Leenstra (Sieger); W.P. Vandertop (Peter); A. Bardelli (Alberto); C.J.F. van Noorden (Cornelis); F.E. Bleeker (Fonnet)

    2014-01-01

    textabstractBackground. Genetic and epigenetic profiling of glioblastomas has provided a comprehensive list of altered cancer genes of which only O6- methylguanine-methyltransferase (MGMT) methylation is used thus far as a predictive marker in a clinical setting. We investigated the prognostic signi

  17. Bioassay and biochemical studies of the status of pirimiphos-methyl and cypermethrin resistance in Aedes (Stegomyia) aegypti and Aedes (Stegomyia) albopictus (Diptera: Culicidae) in Singapore.

    Science.gov (United States)

    Lee, R M L; Choong, C T H; Goh, B P L; Ng, L C; Lam-Phua, S G

    2014-12-01

    Aedes (Stegomyia) aegypti (Linnaeus) and Ae. (Stegomyia) albopictus (Skuse) were sampled from five regions of Singapore (Central, North East, North West, South East and South West) and tested with diagnostic concentrations of the technical grade insecticides, pirimiphos-methyl and cypermethrin. Biochemical assays were performed on the same populations of Ae. aegypti and Ae. albopictus to determine activities of detoxifying enzymes, including non-specific esterase (EST), monooxygenase (MFO) and acetylcholinesterase (AChE). The diagnostic test showed that all Ae. aegypti populations were susceptible to pirimiphos-methyl (mortality = 99 to 100%), but resistant to cypermethrin (mortality = 11 to 76%). Resistance to pirimiphos-methyl was observed in all Ae. albopictus populations (mortality = 49 to 74%) while cypermethrin resistance was detected in most Ae. albopictus populations (mortality = 40 to 75%), except those from Central (mortality = 86%) and South East (mortality = 94%) showing incipient resistance. The biochemical assays showed that there was significant enhancement (P aegypti populations. The biochemical assay results suggested that AChE could play a role in pirimiphos-methyl resistance of Ae. albopictus in South West, South East and North East regions. The small but significant increase in EST activities in Ae. aegypti from all regions suggest that it may play a role in the observed cypermethrin resistance.

  18. Epigenetic inactivation of SPINT2 is associated with tumor suppressive function in esophageal squamous cell carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Yue, Dongli [The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); The Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); Fan, Qingxia [The Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); Chen, Xinfeng; Li, Feng [The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); Wang, Liping [The Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); Huang, Lan [The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); Dong, Wenjie; Chen, Xiaoqi [The Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); Zhang, Zhen [The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); Liu, Jinyan; Wang, Fei [The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); The School of Life Sciences, Zhengzhou University, Zhengzhou 450052, Henan (China); Wang, Meng [The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); The Department of Gastroenterology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); Zhang, Bin [The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan (China); The Department of Hematology/Oncology, School of Medicine, Northwestern University, Chicago 60611 (United States); and others

    2014-03-10

    Hepatocyte growth factor activator inhibitor type 2 (SPINT2), a Kunitz-type serine proteinase inhibitor, has been identified as a putative tumor suppressor gene silenced by promoter methylation. We aimed to investigate whether SPINT2 might act as an esophageal squamous cell carcinoma (ESCC) tumor suppressor gene. Four ESCC cell lines, Fifty-two ESCC tissues and twenty-nine neighboring non-cancerous tissues were included in this study. The expression of SPINT2 was monitored by real time PCR. Bisulfite genomic sequencing and methylation-specific PCR were used to analyze methylation status. The effect of SPINT2 on cell proliferation and apoptosis in EC109 and EC9706 cells was observed by CCK-8 assay and flow cytometric analysis. We found that silencing of SPINT2 was associated with promoter methylation in ESCC cell lines. The densely methylated SPINT2 promoter region was confirmed by bisulfite genomic sequencing. Ectopic expression of SPINT2 inhibited cell proliferation through inducing cell apoptosis in vitro. Furthermore, methylation-specific PCR analysis revealed that SPINT2 promoter methylation was prominent in carcinoma tissues (52.08%) compared with neighboring non-cancerous tissues (22.58%). Kaplan–Meier analysis showed that patients with SPINT2 hypermethylation had shorter survival time. The tumor suppressor gene of SPINT2 is commonly silenced by promoter hypermethylation in human ESCC and SPINT2 hypermethylation is correlated with poor overall survival, implicating SPINT2 is an underlying prognostic marker for human ESCC. - Highlights: • We firstly found SPINT2 gene may be transcriptionally repressed by promoter hypermethylation in ESCC cells. • SPINT2 overexpressing cells induced proliferation inhibition through promoting apoptosis. • mRNA expression of SPINT2 was significantly higher in ESCC tissues than in neighboring non-cancerous tissues. • Promoter hypermethylation of SPINT2 is significantly linked to TNM stage and poor overall survival.

  19. Association between promoter methylation of DAPK gene and HNSCC: A meta-analysis

    Science.gov (United States)

    Cai, Fucheng; Xiao, Xiyue; Niu, Xun; Zhong, Yi

    2017-01-01

    Background The death-associated protein kinase (DAPK) is a tumor suppressor gene, which is a mediator of cell death of INF-γ–induced apoptosis. Aberrant methylation of DAPK promoter has been reported in patients with head and neck squamous cell carcinoma (HNSCC). However, the results of these studies are inconsistent. Hence, the present study aimed to evaluate the association between the promoter methylation of DAPK gene and HNSCC. Methods Relevant studies were systematically searched in PubMed, Web of Science, Ovid, and Embase. The association between DAPK promoter methylation and HNSCC was assessed by odds ratio (ORs) and 95% confidence intervals (CI). To evaluate the potential sources of heterogeneity, we conducted the meta-regression analysis and subgroup analysis. Results Eighteen studies were finally included in the meta-analysis. The frequency of DAPK promoter methylation in patients with HNSCC was 4.09-fold higher than the non-cancerous controls (OR = 3.96, 95%CI = 2.26–6.95). A significant association between DAPK promoter methylation and HNSCC was found among the Asian region and the Non-Asia region (Asian region, OR = 4.43, 95% CI = 2.29–8.58; Non-Asia region, OR = 3.39, 95% CI = 1.18–9.78). In the control source, the significant association between DAPK promoter methylation and HNSCC was seen among the autologous group and the heterogeneous group (autologous group, OR = 2.71, 95% CI = 1.49–4.93; heterogeneous group, OR = 9.50, 95% CI = 2.98–30.27). DAPK promoter methylation was significantly correlated with alcohol status (OR = 1.85, 95% CI = 1.07–3.21). Conclusion The results of this meta-analysis suggested that aberrant methylation of DAPK promoter was associated with HNSCC. PMID:28249042

  20. 食管癌患者肿瘤组织及外周血中上皮细胞钙黏蛋白及Ras相关区域家族1A基因甲基化的研究%DNA methylation of E-cadherin and Ras association domain family 1A in peripheral blood and tumor tissue in patients with esophageal squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    王长春; 毛伟敏; 凌志强

    2012-01-01

    目的 探讨血浆上皮细胞钙黏蛋白( CDH1)基因及Ras相关区域家族1A基因( RASSF1A)甲基化对食管癌手术彻底程度的参考价值及其与食管癌临床病理特征的关系.方法 应用Real-time甲基化特异性聚合酶联反应技术(MSP)技术对76例食管鳞癌患者肿瘤组织、配对的癌旁正常组织、术前、术中及术后外周血游离DNA中CDH1及RASSF1A基因的甲基化状态进行检测.结果 CDH1及RASSF1A甲基化率在食管癌肿瘤组织中分别是72.37%、55.26%;在血浆中分别是63.16%、50.00%,且与病理分期、淋巴结转移及神经脉管浸润显著相关(P值均<0.01);术前、术中、术后血浆中DNA甲基化发生率呈现先升后降的变化趋势.结论 食管鳞癌患者外周血游离DNA中CDH1及RASSF1A甲基化可反映机体的荷瘤状态,并可作为食管鳞癌手术彻底程度的标志物.%Objective To investigate the relationships between E-cadherin (CDH1) and Ras association domain family 1A (RASSF1A) methylation in peripheral blood and clinicopathologic character of esophageal squamous cell carcinoma (ESCC).Methods Real-time methylation specific polymerease chain reaction (MSP) technique was used to detect methylation status of CDH1 and RASSF1A in tumour tissue,adjacent normal tissue and peripheral blood preoperatively in 76 cases of ESCC.Results CDH1 and RASSF1A methylation rate was 72.37% and 55.26% in tumour tissue,and 63.16% and 50.00% in peripheral blood,respectively,and was significantly related to pathology stage,lymph nodes metastasis and nerve vessel invasion ( all P < 0.01 ).DNA methylation rates in peripheral blood was increased initially and then decreased preoperatively,intraoperatively and postoperatively.Conclusion CDH1 and RASSF1 A methylation in peripheral blood in patients with ESCC could reflect the tumor-bearing state of bodies.And CDH1 and RASSF1 A mathylation may be a valuable diagnostic tool as a tumor marker for resection

  1. Ovarian carcinomas with genetic and epigenetic BRCA1 loss have distinct molecular abnormalities

    Directory of Open Access Journals (Sweden)

    Miller Dianne M

    2008-01-01

    Full Text Available Background Subclassification of ovarian carcinomas can be used to guide treatment and determine prognosis. Germline and somatic mutations, loss of heterozygosity (LOH, and epigenetic events such as promoter hypermethylation can lead to decreased expression of BRCA1/2 in ovarian cancers. The mechanism of BRCA1/2 loss is a potential method of subclassifying high grade serous carcinomas. Methods A consecutive series of 49 ovarian cancers was assessed for mutations status of BRCA1 and BRCA2, LOH at the BRCA1 and BRCA2 loci, methylation of the BRCA1 promoter, BRCA1, BRCA2, PTEN, and PIK3CA transcript levels, PIK3CA gene copy number, and BRCA1, p21, p53, and WT-1 immunohistochemistry. Results Eighteen (37% of the ovarian carcinomas had germline or somatic BRCA1 mutations, or epigenetic loss of BRCA1. All of these tumours were high-grade serous or undifferentiated type. None of the endometrioid (n = 5, clear cell (n = 4, or low grade serous (n = 2 carcinomas showed loss of BRCA1, whereas 47% of the 38 high-grade serous or undifferentiated carcinomas had loss of BRCA1. It was possible to distinguish high grade serous carcinomas with BRCA1 mutations from those with epigenetic BRCA1 loss: tumours with BRCA1 mutations typically had decreased PTEN mRNA levels while those with epigenetic loss of BRCA1 had copy number gain of PIK3CA. Overexpression of p53 with loss of p21 expression occurred significantly more frequently in high grade serous carcinomas with epigenetic loss of BRCA1, compared to high grade serous tumors without loss of BRCA1. Conclusion High grade serous carcinomas can be subclassified into three groups: BRCA1 loss (genetic, BRCA1 loss (epigenetic, and no BRCA1 loss. Tumors in these groups show distinct molecular alterations involving the PI3K/AKT and p53 pathways.

  2. Ovarian carcinomas with genetic and epigenetic BRCA1 loss have distinct molecular abnormalities

    Energy Technology Data Exchange (ETDEWEB)

    Gilks, C. Blake; Press, Joshua Z.; De Luca, Alessandro; Boyd, Niki; Young, Sean; Troussard, Armelle; Ridge, Yolanda; Kaurah, Pardeep; Kalloger, Steve E.; Blood, Katherine A.; Smith, Margaret; Spellman, Paul T.; Wang, Yuker; Miller, Dianne M.; Horsman, Doug; Faham, Malek; Gilks, C. Blake; Gray, Joe; Huntsman, David G.

    2008-05-02

    Subclassification of ovarian carcinomas can be used to guide treatment and determine prognosis. Germline and somatic mutations, loss of heterozygosity (LOH), and epigenetic events such as promoter hypermethylation can lead to decreased expression of BRCA1/2 in ovarian cancers. The mechanism of BRCA1/2 loss is a potential method of subclassifying high grade serous carcinomas. A consecutive series of 49 ovarian cancers was assessed for mutations status of BRCA1 and BRCA2, LOH at the BRCA1 and BRCA2 loci, methylation of the BRCA1 promoter, BRCA1, BRCA2, PTEN, and PIK3CA transcript levels, PIK3CA gene copy number, and BRCA1, p21, p53, and WT-1 immunohistochemistry. Eighteen (37%) of the ovarian carcinomas had germline or somatic BRCA1 mutations, or epigenetic loss of BRCA1. All of these tumors were high-grade serous or undifferentiated type. None of the endometrioid (n=5), clear cell (n=4), or low grade serous (n=2) carcinomas showed loss of BRCA1, whereas 47% of the 38 high-grade serous or undifferentiated carcinomas had loss of BRCA1. It was possible to distinguish high grade serous carcinomas with BRCA1 mutations from those with epigenetic BRCA1 loss: tumors with BRCA1 mutations typically had decreased PTEN mRNA levels while those with epigenetic loss of BRCA1 had copy number gain of PIK3CA. Overexpression of p53 with loss of p21 expression occurred significantly more frequently in high grade serous carcinomas with epigenetic loss of BRCA1, compared to high grade serous tumors without loss of BRCA1. High grade serous carcinomas can be subclassified into three groups: BRCA1 loss (genetic), BRCA1 loss (epigenetic), and no BRCA1 loss. Tumors in these groups show distinct molecular alterations involving the PI3K/AKT and p53 pathways.

  3. Ovarian carcinomas with genetic and epigenetic BRCA1 loss havedistinct molecular abnormalities

    Energy Technology Data Exchange (ETDEWEB)

    Press, Joshua Z.; De Luca, Alessandro; Boyd, Niki; Young, Sean; Troussard, Armelle; Ridge, Yolanda; Kaurah, Pardeep; Kalloger, Steve E.; Blood, Katherine A.; Smith, Margaret; Spellman, Paul T.; Wang, Yuker; Miller, Dianne M.; Horsman, Doug; Faham, Malek; Gilks, C. Blake; Gray,Joe; Huntsman, David G.

    2007-07-23

    Subclassification of ovarian carcinomas can be used to guide treatment and determine prognosis. Germline and somatic mutations, loss of heterozygosity (LOH), and epigenetic events such as promoter hypermethylation can lead to decreased expression of BRCA1/2 in ovarian cancers. The mechanism of BRCA1/2 loss is a potential method of subclassifying high grade serous carcinomas. A consecutive series of 49 ovarian cancers was assessed for mutations status of BRCA1 and BRCA2, LOH at the BRCA1 and BRCA2 loci, methylation of the BRCA1 promoter, BRCA1, BRCA2, PTEN, and PIK3CA transcript levels, PIK3CA gene copy number, and BRCA1, p21, p53, and WT-1 immunohistochemistry. Eighteen (37%) of the ovarian carcinomas had germline or somatic BRCA1 mutations, or epigenetic loss of BRCA1. All of these tumors were high-grade serous or undifferentiated type. None of the endometrioid (n = 5), clear cell (n = 4), or low grade serous (n = 2) carcinomas showed loss of BRCA1, whereas 47% of the 38 high-grade serous or undifferentiated carcinomas had loss of BRCA1. It was possible to distinguish high grade serous carcinomas with BRCA1 mutations from those with epigenetic BRCA1 loss: tumors with BRCA1 mutations typically had decreased PTEN mRNA levels while those with epigenetic loss of BRCA1 had copy number gain of PIK3CA. Overexpression of p53 with loss of p21 expression occurred significantly more frequently in high grade serous carcinomas with epigenetic loss of BRCA1, compared to high grade serous tumors without loss of BRCA1. High grade serous carcinomas can be subclassified into three groups: BRCA1 loss (genetic), BRCA1 loss (epigenetic), and no BRCA1 loss. Tumors in these groups show distinct molecular alterations involving the PI3K/AKT and p53 pathways.

  4. Methylation-Specific PCR Unraveled

    Directory of Open Access Journals (Sweden)

    Sarah Derks

    2004-01-01

    Full Text Available Methylation‐specific PCR (MSP is a simple, quick and cost‐effective method to analyze the DNA methylation status of virtually any group of CpG sites within a CpG island. The technique comprises two parts: (1 sodium bisulfite conversion of unmethylated cytosine's to uracil under conditions whereby methylated cytosines remains unchanged and (2 detection of the bisulfite induced sequence differences by PCR using specific primer sets for both unmethylated and methylated DNA. This review discusses the critical parameters of MSP and presents an overview of the available MSP variants and the (clinical applications.

  5. MGMT promoter methylation status and MGMT and CD133 immunohistochemical expression as prognostic markers in glioblastoma patients treated with temozolomide plus radiotherapy

    OpenAIRE

    Melguizo Consolación; Prados Jose; González Beatriz; Ortiz Raul; Concha Angel; Alvarez Pablo; Madeddu Roberto; Perazzoli Gloria; Oliver Jaime; López Rodrigo; Rodríguez-Serrano Fernando; Aránega Antonia

    2012-01-01

    Abstract Background The CD133 antigen is a marker of radio- and chemo-resistant stem cell populations in glioblastoma (GBM). The O6-methylguanine DNA methyltransferase (MGMT) enzyme is related with temozolomide (TMZ) resistance. Our propose is to analyze the prognostic significance of the CD133 antigen and promoter methylation and protein expression of MGMT in a homogenous group of GBM patients uniformly treated with radiotherapy and TMZ. The possible connection between these GBM markers was ...

  6. DNA methylation and demethylation:current status and future per-spective%DNA甲基化和去甲基化的研究现状及思考

    Institute of Scientific and Technical Information of China (English)

    邓大君

    2014-01-01

    DNA methylation plays important roles in cell differentiation, embryonic development, host adaptations to environmental factors, and pathogenesis through regulation of gene transcription and imprinting, X-inactivation, and de-fense of foreign genetic material invasion, is currently one of the hottest research fields on epigenetics. In the past few years, a number of important findings on DNA methylation have been achieved. These findings include discovery of TETs-catalyzed cytosine hydroxymethylation and its functions in the early embryonic development; the relationship be-tween active and passive DNA demethylation;establishment and maintenance of DNA methylation patterns and their asso-ciations with histone modifications, chromatin configuration, polycomb group proteins and non-coding RNA bindings. DNA methylation has become a new potential biomarker and therapy target.%DNA甲基化通过调节基因转录、印记、X染色体灭活和防御外源性遗传物质入侵等,在细胞分化、胚胎发育、环境适应和疾病发生发展上发挥重要作用,是当前表观遗传学研究的热点领域之一。文章介绍了在过去几年中TET介导的DNA羟甲基化及其在早期胚胎发育中的作用, DNA主动去甲基化及其与被动去甲基化的关系, DNA甲基化建立及其与组蛋白修饰、染色质构象、多梳蛋白和非编码RNA结合等关系方面的重要研究进展和存在的问题以及DNA甲基化的转化应用前景。

  7. DNA Methylation

    OpenAIRE

    Alokail, Majed S.; Alenad, Amal M.

    2015-01-01

    The DNA of E. coli contains 19,120 6-methyladenines and 12,045 5-methylcytosines in addition to the four regular bases and these are formed by the postreplicative action of three DNA methyltransferases. The majority of the methylated bases are formed by the Dam and Dcm methyltransferases encoded by the dam (DNA adenine methyltransferase) and dcm (DNA cytosine methyltransferase) genes. Although not essential, Dam methylation is important for strand discrimination during repair of replication e...

  8. The DNA methylation events in normal and cloned rabbit embryos

    Institute of Scientific and Technical Information of China (English)

    TaoChen; Yan-LingZhang; YanJiang; Shu-ZhenLiu; HeideSchatten; Da-YuanChen; Qing-YuanSun

    2005-01-01

    To study the DNA methylation events in normal and cloned rabbit embryos, we investigated the methylation status of a satellite seqnence and the promoter region of a single-copy gene using bisulfite-sequencing technology. During normal rabbit embryo development, both sequences maintained hypermethylation status until the 8- to 16-cell stage when progressive demethylation took place. In cloned embryos, the single-copy gene promoter sequence was rapidly demethylated and preco-ciously de novo methylated, while the satellite sequence mainrained the donor-type methylation status in all examined stages. Our results indicate that unique sequences as well as satellitesequences may have aberrant methylation patterns in cloned embryos.

  9. Estrogen and progesterone receptor status determined by the Ventana ES 320 automated immunohistochemical stainer and the CAS 200 image analyzer in 236 early-stage breast carcinomas: prognostic significance.

    Science.gov (United States)

    Layfield, L J; Saria, E A; Conlon, D H; Kerns, B J

    1996-03-01

    The quantitation of estrogen and progesterone receptors (ER and PgR) has become the standard of care in the evaluation of patients with primary breast carcinoma. It has been demonstrated that ER and PgR detected by immunohistochemical methods in formalin-fixed paraffin-embedded tissue can be quantified by computerized image analysis. In this study, ER and PgR levels were determined by using an automated immunochemistry stainer (Ventana ES 320) and an image analyzer (CAS 200) in a series of 236 patients with stage I/II carcinoma of the breast. The degree of correlation of the ER and PgR levels determined by the dextran-coated charcoal method (DCC) with image analysis quantitation was high (r=0.75). The agreement between both methods was 77% for ER and 73% for PgR. Hormone receptor levels were correlated with prognosis as determined by overall survival. An ER level of 30 fmol/mg as determined by image analysis was established to stratify the patient population most effectively into favorable and unfavorable prognostic groups (P=0.003). An ER level of 20 fmol/mg for prognostic stratification reached statistical significance (P=0.03). The DCC method was not able to stratify the patients into prognostic groups at the traditionally accepted cutpoint of 10 fmol/mg (P=0.52). We conclude that when used in combination, automated immunohistochemistry and quantitative image analysis offer a favorable alternative to the DCC method in assessment of ER and PgR status in human mammary carcinoma. In addition, quantitative immunocytochemistry techniques may prove superior to the DCC method in specimens in which there is limited tumor volume (including fine-needle aspirates), stroma-rich tumors, and early-stage lesions including intraductal carcinoma.

  10. DNA methylation

    DEFF Research Database (Denmark)

    Williams, Kristine; Christensen, Jesper; Helin, Kristian

    2012-01-01

    DNA methylation is involved in key cellular processes, including X-chromosome inactivation, imprinting and transcriptional silencing of specific genes and repetitive elements. DNA methylation patterns are frequently perturbed in human diseases such as imprinting disorders and cancer. The recent...... discovery that the three members of the TET protein family can convert 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC) has provided a potential mechanism leading to DNA demethylation. Moreover, the demonstration that TET2 is frequently mutated in haematopoietic tumours suggests that the TET...... proteins are important regulators of cellular identity. Here, we review the current knowledge regarding the function of the TET proteins, and discuss various mechanisms by which they contribute to transcriptional control. We propose that the TET proteins have an important role in regulating DNA methylation...

  11. Effects of 5-aza-2′deoxycytidine on RECK gene expression and tumor invasion in salivary adenoid cystic carcinoma

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, X.Q. [Department of Oral and Maxillofacial Surgery, School of Stomatology, Shandong University, Jinan (China); Department of Oral and Maxillofacial Surgery, The First People' s Hospital of Jining, Shandong (China); Department of Oral and Maxillofacial Surgery, Shandong Provincial Hospital, Affiliated to Shandong University, Jinan (China); Huang, S.Y. [Department of Oral and Maxillofacial Surgery, Shandong Provincial Hospital, Affiliated to Shandong University, Jinan (China); Zhang, D.S. [Department of Oral and Maxillofacial Surgery, School of Stomatology, Shandong University, Jinan (China); Department of Oral and Maxillofacial Surgery, Shandong Provincial Hospital, Affiliated to Shandong University, Jinan (China); Zhang, S.Z.; Li, W.G.; Chen, Z.W.; Wu, H.W. [Department of Oral and Maxillofacial Surgery, Shandong Provincial Hospital, Affiliated to Shandong University, Jinan (China)

    2014-12-12

    Reversion-inducing cysteine-rich protein with kazal motifs (RECK), a novel tumor suppressor gene that negatively regulates matrix metalloproteinases (MMPs), is expressed in various normal human tissues but downregulated in several types of human tumors. The molecular mechanism for this downregulation and its biological significance in salivary adenoid cystic carcinoma (SACC) are unclear. In the present study, we investigated the effects of a DNA methyltransferase (DNMT) inhibitor, 5-aza-2′deoxycytidine (5-aza-dC), on the methylation status of the RECK gene and tumor invasion in SACC cell lines. Methylation-specific PCR (MSP), Western blot analysis, and quantitative real-time PCR were used to investigate the methylation status of the RECK gene and expression of RECK mRNA and protein in SACC cell lines. The invasive ability of SACC cells was examined by the Transwell migration assay. Promoter methylation was only found in the ACC-M cell line. Treatment of ACC-M cells with 5-aza-dC partially reversed the hypermethylation status of the RECK gene and significantly enhanced the expression of mRNA and protein, and 5-aza-dC significantly suppressed ACC-M cell invasive ability. Our findings showed that 5-aza-dC inhibited cancer cell invasion through the reversal of RECK gene hypermethylation, which might be a promising chemotherapy approach in SACC treatment.

  12. A lower degree of PBMC L1 methylation in women with lower folate status may explain the MTHFR C677T polymorphism associated higher risk of CIN in the US post folic acid fortification era.

    Directory of Open Access Journals (Sweden)

    Suguna Badiga

    Full Text Available Studies in populations unexposed to folic acid (FA fortification have demonstrated that MTHFR C677T polymorphism is associated with increased risk of higher grades of cervical intraepithelial neoplasia (CIN 2+. However, it is unknown whether exposure to higher folate as a result of the FA fortification program has altered the association between MTHFR C677T and risk of CIN, or the mechanisms involved with such alterations. The current study investigated the following in a FA fortified population: 1 The association between MTHFR C677T polymorphism and risk of CIN 2+; 2 The modifying effects of plasma folate concentrations on this association; and 3 The modifying effects of plasma folate on the association between the polymorphism and degree of methylation of long interspersed nucleotide elements (L1s, in peripheral blood mononuclear cell (PBMC DNA, a documented biomarker of CIN risk.The study included 457 US women diagnosed with either CIN 2+ (cases or ≤ CIN 1 (non-cases. Unconditional logistic regression models were used to test the associations after adjusting for relevant risk factors for CIN.The 677CT/TT MTHFR genotypes were not associated with the risk of CIN 2+. Women with CT/TT genotype with lower folate, however, were more likely to be diagnosed with CIN 2+ compared to women with CT/TT genotype with higher folate (OR = 2.41, P = 0.030. Women with CT/TT genotype with lower folate were less likely to have a higher degree of PBMC L1 methylation compared to women with CT/TT genotype with higher folate (OR = 0.28, P = 0.017.This study provides the first evidence that the MTHFR 677CT/TT genotype-associated lower degree of PBMC L1 methylation increases the risk of CIN 2+ in women in the US post-FA fortification era. Thus, even in the post-FA fortification era, not all women have adequate folate status to overcome MTHFR 677CT/TT genotype-associated lower degree of L1 methylation.

  13. 睾丸组织中的TSP50启动子甲基化差异的研究%Difference in TSP50 promoter methylation status of testis tissue

    Institute of Scientific and Technical Information of China (English)

    茅淑砚; 杨明; 蒲春雪; 吕静静; 王悦增

    2011-01-01

    目的:检测不同组织中TSP50启动子区域的甲基化状态,研究其表达的机制.方法:分离、纯化鼠睾丸中的精原细胞、精母细胞和精子,分别提取人的睾丸组织、Ntera-2细胞、鼠精原细胞、精母细胞和精子的基因组DNA,用重亚硫酸盐处理基因组DNA,PCR扩增人和鼠的TSP50的启动子CpG岛区域,电泳,提纯,连接于质粒中,转导入细胞中,扩增,提取,纯化质粒,测序.结果:人睾丸组织甲基化水平较低,Ntera-2细胞处于较高水平.鼠的精原细胞和精子细胞甲基化水平较高,精母细胞甲基化水平较低.结论:TSP50启动子区域甲基化位点的甲基化水平调控TSP50蛋白的表达.%Objective:To detect the methylation status of the promoter of TSP50 in different tissues and explore its expression mechanism.Methods:To iso late and purify the spem atogonias ,apem atocytes and spem atozoa of mouse tesis, and extract the genome DNA of hum an testis tissue,N tera-2 cells and spem atogonia,spem atocyte,spem atozoa of mouse respectively.Genome DNA was treated with bisulfite,am plificate the CPG region of the promoter of TSP50 of hum an and mouse by PCR.Electrophoresis,purify and link them into T vector,and then transduce them into DH 5α cells to am plificate,exact and purify the plasm ids and to sequence the CPG region of the promoter of TSP50.Results:The methylation level is low in the hum an tesis but at a higher level in the tera-2 cells.As to mouse,the methylation level is high in the spem atogonias and the spem atoblasts but is at a low level in the apem atocytes.Conclusion:The methyaltion level of the methylation site of the promoter region of TSP50 regulates the expression of TSP50 protein.

  14. Expression of SOCS-3 and Its Methylation in Renal Cancer

    Institute of Scientific and Technical Information of China (English)

    YU Jing; ZHAO Li-chun; YAO Song-mei; HE Cheng-yan; LI Hong-jun; KONG Xiang-bo

    2011-01-01

    The expression level of suppressor of cytokine signaling-3(SOCS-3) in human renal carcinoma and its methylation state were investigated. Reverse transcription-polymerase chain reaction(RT-PCR), immuocytochemistry,immunohistochemistry and Western blot were used to detect the expression level of SOCS-3, and the methylation of SOCS gene was investigated by methylation specific PCR in the tissues of i 5 cases of renal carcinoma. Compared to those of the normal renal cell line and specimens, the expression level of SOCS-3 in renal carcinoma was significantly lower or can't be detected(P<0.01). And the methylation of SOCS-3 gene in the tissue of renal carcinoma was significantly higher. The expression of SOCS-3 gene is significantly lower in renal carcinoma and the high methylation of the promoter island of SOCS-3 gene is associated with the lower expression of SOCS-3 gene. It may be one of main mechanisms for the development and progress of renal carcinoma.

  15. Methylated genes as new cancer biomarkers.

    LENUS (Irish Health Repository)

    Duffy, M J

    2012-02-01

    Aberrant hypermethylation of promoter regions in specific genes is a key event in the formation and progression of cancer. In at least some situations, these aberrant alterations occur early in the formation of malignancy and appear to be tumour specific. Multiple reports have suggested that measurement of the methylation status of the promoter regions of specific genes can aid early detection of cancer, determine prognosis and predict therapy responses. Promising DNA methylation biomarkers include the use of methylated GSTP1 for aiding the early diagnosis of prostate cancer, methylated PITX2 for predicting outcome in lymph node-negative breast cancer patients and methylated MGMT in predicting benefit from alkylating agents in patients with glioblastomas. However, prior to clinical utilisation, these findings require validation in prospective clinical studies. Furthermore, assays for measuring gene methylation need to be standardised, simplified and evaluated in external quality assurance programmes. It is concluded that methylated genes have the potential to provide a new generation of cancer biomarkers.

  16. 人脑胶质瘤细胞中GDNF基因启动子1区甲基化状态的研究%Study of the Methylation Status of Promoter 1 Region of GDNF Gene in Human Glioma Cells

    Institute of Scientific and Technical Information of China (English)

    任庆先; 王建村; 王莉; 高殿帅; 虞正权

    2012-01-01

    Objective: To observe the methylation status of GDNF promoter 1 region in human glioma, in order to explore the effect of methylation on the expression of GDNF in glioma. Methods: Genesequencing detected the GDNF gene order in 10 cases of glioma and 5 normal brain tissues, to compare whether their genetic mutations have occurred; the methylation modified status of GDNF promoter 1 region in 20 cases of glioma (10 cases of low-level and 10 cases of high-level) and 5 cases of normal brain tissues were detected through genesequencing after bisulfite modification. Results: The mutations of GDNF gene in promoter 1 region were not observed in glioma; the methylation of GDNF gene in promoter 1 region in normal brain tissues, low-level and high-level glioma were 72.25%, 86.25%, 86.75% respectively. The methylation level in glioma was significantly higher when compared to normal brain tissues (P<0.05); while there was no significant differences in high-and low-level tissues. Conclusions: Hypermethylation occuerred in GDNF promoter 1 region might influence the expression of GDNF in glioma cell.%目的:观察GDNF启动子1区在人脑胶质瘤细胞中的甲基化修饰状态,以期探讨其对于GDNF在胶质瘤中高表达的影响.方法:基因测序检测10例胶质瘤与5例正常脑组织中GDNF基因序列,比较其基因是否有突变发生;重亚硫酸盐修饰后基因测序检测20例胶质瘤(10例低级别和10例高级别)与5例正常脑组织中GDNF启动子1区甲基化修饰状态.结果:GDNF启动子1区基因在胶质瘤中没有发生突变;GDNF启动子1区甲基化修饰在正常脑组织、低级别、高级别中发生率分别为72.25%、86.25%、86.75%.在胶质瘤中的甲基化修饰水平比正常脑组织明显增高(P<0.05),而高低级别之间无显著性差异.结论:在胶质瘤细胞中,GDNF启动子1区发生了高甲基化修饰,这种修饰很可能会影响GDNF基因的表达.

  17. Marked Differences in C9orf72 Methylation Status and Isoform Expression between C9/ALS Human Embryonic and Induced Pluripotent Stem Cells

    Directory of Open Access Journals (Sweden)

    Yaara Cohen-Hadad

    2016-11-01

    Full Text Available We established two human embryonic stem cell (hESC lines with a GGGGCC expansion in the C9orf72 gene (C9, and compared them with haploidentical and unrelated C9 induced pluripotent stem cells (iPSCs. We found a marked difference in C9 methylation between the cells. hESCs and parental fibroblasts are entirely unmethylated while the iPSCs are hypermethylated. In addition, we show that the expansion alters promoter usage and interferes with the proper splicing of intron 1, eventually leading to the accumulation of repeat-containing mRNA following neural differentiation. These changes are attenuated in C9 iPSCs, presumably owing to hypermethylation. Altogether, this study highlights the importance of neural differentiation in the pathogenesis of disease and points to the potential role of hypermethylation as a neuroprotective mechanism against pathogenic mRNAs, envisaging a milder phenotype in C9 iPSCs.

  18. Histone H3 Acetyl K9 and Histone H3 Tri Methyl K4 as Prognostic Markers for Patients with Cervical Cancer.

    Science.gov (United States)

    Beyer, Susanne; Zhu, Junyan; Mayr, Doris; Kuhn, Christina; Schulze, Sandra; Hofmann, Simone; Dannecker, Christian; Jeschke, Udo; Kost, Bernd P

    2017-02-23

    Chromatin remodeling alters gene expression in carcinoma tissue. Although cervical cancer is the fourth most common cancer in women worldwide, a systematic study about the prognostic value of specific changes in the chromatin structure, such as histone acetylation or histone methylation, is missing. In this study, the expression of histone H3 acetyl K9, which is known to denote active regions at enhancers and promoters, and histone H3 tri methyl K4, which preferentially identifies active gene promoters, were examined as both show high metastatic potential. A panel of patients with cervical cancer was selected and the importance of the histone modifications concerning survival-time (overall survival and relapse-free survival) was analyzed in 250 cases. Histone H3 acetyl K9 staining was correlated with low grading, low FIGO (TNM classification and the International Federation of Gynecology and Obstetrics) status, negative N-status and low T-status in cervical cancer, showing a higher expression in adenocarcinoma than in squamous cell carcinoma. Cytoplasmic expression of histone H3 tri methyl K4 in a cervical cancer specimen was correlated with advanced T-status and poor prognosis. While cytoplasmic H3K4me3 expression seemed to be a marker of relapse-free survival, nuclear expression showed a correlation to poor prognosis in overall survival. Within this study, we analyzed the chemical modification of two histone proteins that are connected to active gene expression. Histone H3 acetyl K9 was found to be an independent marker of overall survival. Histone H3 tri methyl K4 was correlated with poor prognosis and it was found to be an independent marker of relapse-free survival. Therefore, we could show that chromatin remodeling plays an important role in cervical cancer biology.

  19. Histone H3 Acetyl K9 and Histone H3 Tri Methyl K4 as Prognostic Markers for Patients with Cervical Cancer

    Directory of Open Access Journals (Sweden)

    Susanne Beyer

    2017-02-01

    Full Text Available Chromatin remodeling alters gene expression in carcinoma tissue. Although cervical cancer is the fourth most common cancer in women worldwide, a systematic study about the prognostic value of specific changes in the chromatin structure, such as histone acetylation or histone methylation, is missing. In this study, the expression of histone H3 acetyl K9, which is known to denote active regions at enhancers and promoters, and histone H3 tri methyl K4, which preferentially identifies active gene promoters, were examined as both show high metastatic potential. A panel of patients with cervical cancer was selected and the importance of the histone modifications concerning survival-time (overall survival and relapse-free survival was analyzed in 250 cases. Histone H3 acetyl K9 staining was correlated with low grading, low FIGO (TNM classification and the International Federation of Gynecology and Obstetrics status, negative N-status and low T-status in cervical cancer, showing a higher expression in adenocarcinoma than in squamous cell carcinoma. Cytoplasmic expression of histone H3 tri methyl K4 in a cervical cancer specimen was correlated with advanced T-status and poor prognosis. While cytoplasmic H3K4me3 expression seemed to be a marker of relapse-free survival, nuclear expression showed a correlation to poor prognosis in overall survival. Within this study, we analyzed the chemical modification of two histone proteins that are connected to active gene expression. Histone H3 acetyl K9 was found to be an independent marker of overall survival. Histone H3 tri methyl K4 was correlated with poor prognosis and it was found to be an independent marker of relapse-free survival. Therefore, we could show that chromatin remodeling plays an important role in cervical cancer biology.

  20. Elderly patients at higher risk of laryngeal carcinoma recurrence could be identified by a panel of two biomarkers (nm23-H1 and CD105) and pN+ status.

    Science.gov (United States)

    Lovato, Andrea; Marioni, Gino; Manzato, Enzo; Staffieri, Claudia; Giacomelli, Luciano; Ralli, Giovanni; Staffieri, Alberto; Blandamura, Stella

    2015-11-01

    Laryngeal squamous cell carcinoma (LSCC) recurrences are very difficult to manage in elderly patients (age ≥65 years), because treatment carries significant morbidity and mortality. The aim of this study was to develop a panel of parameters (clinicopathological variables or biomarkers) to improve our ability to detect elderly patients at higher risk of LSCC recurrence. Maspin, nm23-H1, and CD105 were investigated using immunohistochemistry on surgical specimens from 46 elderly patients treated for LSCC. After univariate analysis identified parameters associated with LSCC recurrence, a multivariate prognostic model was constructed. At univariate analysis, a higher recurrence rate was significantly associated with nm23-H1 nuclear expression in carcinoma cells ≤2.0% (p = 0.01), CD105 expression in intratumoral vascular endothelial cells ≥5.28% (p = 0.04), and pN+ status (p = 0.04). Multivariate modeling confirmed that nuclear nm23-H1 ≤2.0% (p = 0.009) and CD105 ≥5.28% (p = 0.013) had a negative prognostic significance in terms of disease recurrence, while pN+ status showed a trend toward significance (p = 0.05). We thus obtained a panel comprising two biomarkers and neck lymph node status that revealed an excellent discriminatory power [AUC (ROC) of 0.81] in terms of the risk of LSCC recurrence. The panel achieved a specificity of 96% and a positive predictive value of 93%. We identified a panel with an excellent discriminatory power in identifying elderly patients at higher risk of recurrence after treatment for LSCC. These patients would benefit from a more aggressive primary treatment.

  1. Methylation status in promoter of APC and environmental factors with the predisposition to prostate cancer%APC基因启动子区甲基化及环境因素与前列腺癌关系研究

    Institute of Scientific and Technical Information of China (English)

    张连升; 梁永; 崔飞伦

    2012-01-01

    Objective To investigate the relationship between methylation status of adenomatosis polyposis coli gene ( APC) promoter and environmental factors in the predisposition of prostate cancer (Pea) in Chinese. Methods Bisulphite sequencing PCR were used to detect the expression of APC genes in prostate cancer tissues from 60 patients and hy-perplastic tissues from 40 benign prostate hyperplasia ( BPH) patients. Environmental risk factors related with Pca were also collected including demographic information, body mass index (BMI) , drinking, tea drinking, smoking, sports. Relationship between methylation of APC gene, environmental risk factors and the PCa prevalence was analysised by using uni-variate and multivariate method. Results The methylation rates of CG sites in promoter of APC genes were 14. 00% in PCa and 1. 19% in BPH. Methylation rate in APC genes was associated with PSA, Gleason score, pathologic stage. PCa was correlated with drinking habit of wire (OR = 2.46) or tea ( OR = 0. 29). Conclusions The methylation of promoter in APC genes is related to the development of PCa and the classification of risk. Methylation of APC genes may serve as a promising method for early diagnosis of PCa. The drinking habit of wine or tea is also a risk factor for PCa.%目的 探讨腺瘤样结肠息肉易感基因(APC)启动子区CpG甲基化及环境危险因素与中国人群中前列腺癌(PCa)发病之间的关系.方法 收集60例PCa和40例前列腺增生(BPH)患者的组织标本.用亚硫酸盐修饰后测序法检测PCa组织及BPH组织中APC基因启动子区CpG甲基化情况.同时收集人口学资料以及体质量指数(BMI)、饮酒、饮茶、吸烟、体育运动等环境危险因素资料,用单因素和多因素分析法研究APC基因甲基化及环境危险因素与PCa患病之间的关系.结果 APC基因在PCa及BPH组织中CG位点甲基化率为14.00%、1.19%;基因甲基化率与前列腺特异性抗原(PSA)、Gleason评分、病理分期

  2. AN UPWARD TREND IN DNA P16INK4A METHYLATION PATTERN AND HIGH RISK HPV INFECTION ACCORDING TO THE SEVERITY OF THE CERVICAL LESION

    Directory of Open Access Journals (Sweden)

    Fernanda Nahoum Carestiato

    2013-09-01

    Full Text Available SUMMARY High-risk human papillomavirus (hr-HPV infection is necessary but not sufficient for cervical cancer development. Recently, P16INK4A gene silencing through hypermethylation has been proposed as an important cofactor in cervical carcinogenesis due to its tumor suppressor function. We aimed to investigate P16INK4A methylation status in normal and neoplastic epithelia and evaluate an association with HPV infection and genotype. This cross-sectional study was performed with 141 cervical samples from patients attending Hospital Moncorvo Filho, Rio de Janeiro. HPV detection and genotyping were performed through PCR and P16INK4A methylation by nested-methylation specific PCR (MSP. HPV frequency was 62.4% (88/141. The most common HPV were HPV16 (37%, HPV18 (16.3% and HPV33/45(15.2%. An upward trend was observed concerning P16INK4A methylation and lesion degree: normal epithelia (10.7%, low grade lesions (22.9%, high grade (57.1% and carcinoma (93.1% (p < 0.0001. A multivariate analysis was performed to evaluate an association between methylation, age, tobacco exposure, HPV infection and genotyping. A correlation was found concerning methylation with HPV infection (p < 0.0001, hr-HPV (p = 0.01, HSIL (p < 0.0007 and malignant lesions (p < 0.0001. Since viral infection and epigenetic alterations are related to cervical carcinoma, we suggest that P16INK4A methylation profile maybe thoroughly investigated as a biomarker to identify patients at risk of cancer.

  3. Parotid carcinoma

    DEFF Research Database (Denmark)

    Sørensen, Kristine Bjørndal; Godballe, Christian; de Stricker, Karin;

    2006-01-01

    OBJECTIVES: Our aim is to investigate the expression of kit protein (KIT) and epidermal growth factor receptor (EGFR) in parotid carcinomas in order to correlate the expression to histology and prognosis. Further we want to perform mutation analysis of KIT-positive adenoid cystic carcinomas....... PATIENTS AND METHODS: Formalin-fixed paraffin-embedded sections from 73 patients with parotid gland carcinomas were used for the study. The sections were stained with both KIT and EGFR polyclonal antibodies. Twelve KIT-positive adenoid cystic carcinomas were examined for c-kit mutation in codon 816....... RESULTS: Of all carcinomas 25% were KIT-positive and 79% were EGFR-positive. Ninety-two percentage of the adenoid cystic carcinomas were KIT-positive. None of the adenoid cystic carcinomas had mutations in codon 816 of the c-kit gene. CONCLUSION: Neither KIT- nor EGFR-expression seem to harbour...

  4. Methylation of SLIT2,DAPK,RIZ1,CHFR,EDNRB,3-OST-2 genes in hepatocellular carcinoma cell lines%肝癌细胞系中SLIT2、DAPK、RIZ1、CHFR、EDNRB、3-OST-2基因甲基化状态

    Institute of Scientific and Technical Information of China (English)

    李培坤; 耿小平; 朱立新; 李晓明; 刘付宝; 赵红川

    2011-01-01

    目的 了解SUN449、BEL-7402、SMMC-7721、Hep3B和HepG2 5种肝癌细胞系中SLIT2、DAPK、RIZ1、CHFR、EDNRB和3-OST-2 6种基因启动子区域甲基化状态.方法 体外培养以上5种肝癌细胞系,应用甲基化特异性PCR(MSP)技术,检测5种肝癌细胞系SLIT2、DAPK、RIZ1、CHFR、EDNRB和3-OST-2 6种基因启动子区域的甲基化状态.结果 SLIT2、RIZ1、EDNRB和3-OST-2 4种基因启动子区域在5种肝癌细胞系SUN449、BEL-7402、SMMC-7721、Hep3B和HepG2中为完全甲基化,甲基化率均为100%;DAPK基因在此5种肝癌细胞系中为完全非甲基化,未发现甲基化状态,甲基化率为0; CHFR基因启动子区在SUN449、BEL-7402、SMMC-7721和HepG2细胞系中发生甲基化,甲基化率为80%,在SUN449、SMMC-7721和HepG2中为不完全甲基化,在BEL-7402细胞系中为完全甲基化,在Hep3B中为完全非甲基化.结论 肝癌细胞系中SLIT2、DAPK、RIZ1、CHFR、EDNRB和3-OST-2基因启动子区的高甲基化是普遍现象,而DAPK基因呈低甲基化水平.%Objective To study the methylation of SLIT2DAPK. RIZ1 . CHFR, EDNRB and 3-OST-2 genes in 5 hepatocellular carcinoma( HCC ) cell lines-SUN449, BEL-7402, SMMC-7721 , Hep3B and HepG2. Methods Methylation-specific PCR was adopted to detected the promoter methylation of these genes in the 5 HCC cell lines cultured in vitro. Results SLIT2 , RIZ1 .EDNRB and 3-OST-2 were completely showing methylation in SUN449 . BEL-7402 ,SMMC-7721 ,Hep3B and HepG2. The methylation rate was l00% ; DAPK was completely unmethylation, and the methylation rate was 0; Methylation of CHFR was deteceted in SUN449 . BEL-7402 . SMMC-7721 and HepG2, The methylation rates were 80% . Methylation was deteceted partly in SUN449 , SMMC-772 and HepG2,but completely methylation in BEL-7402 and completely unmethylation in Hep3B. Conclusion Methylation of SLIT2 . DAPK. RIZI .CHFR .EDNRB and 3-OST-2 is a common event in 5 HCC cell lines, but the methylation rate of DAPK is low.

  5. Using peripheral blood circulating DNAs to detect CpG global methylation status and genetic mutations in patients with myelodysplastic syndrome

    Energy Technology Data Exchange (ETDEWEB)

    Iriyama, Chisako [Department of Hematology and Oncology, Nagoya University Graduate School of Medicine, Nagoya (Japan); Tomita, Akihiro, E-mail: atomita@med.nagoya-u.ac.jp [Department of Hematology and Oncology, Nagoya University Graduate School of Medicine, Nagoya (Japan); Hoshino, Hideaki; Adachi-Shirahata, Mizuho [Department of Hematology and Oncology, Nagoya University Graduate School of Medicine, Nagoya (Japan); Furukawa-Hibi, Yoko; Yamada, Kiyofumi [Department of Neuropsychopharmacology and Hospital Pharmacy, Nagoya University School of Medicine, Nagoya (Japan); Kiyoi, Hitoshi; Naoe, Tomoki [Department of Hematology and Oncology, Nagoya University Graduate School of Medicine, Nagoya (Japan)

    2012-03-23

    Highlights: Black-Right-Pointing-Pointer Circulating DNAs (CDs) can be used to detect genetic/epigenetic abnormalities in MDS. Black-Right-Pointing-Pointer Epigenetic changes can be detected more sensitively when using plasma DNA than PBMNC. Black-Right-Pointing-Pointer Mutation ratio in CDs may reflect the ratio in stem cell population in bone marrow. Black-Right-Pointing-Pointer Using CDs can be a safer alternate strategy compared to bone marrow aspiration. -- Abstract: Myelodysplastic syndrome (MDS) is a hematopoietic stem cell disorder. Several genetic/epigenetic abnormalities are deeply associated with the pathogenesis of MDS. Although bone marrow (BM) aspiration is a common strategy to obtain MDS cells for evaluating their genetic/epigenetic abnormalities, BM aspiration is difficult to perform repeatedly to obtain serial samples because of pain and safety concerns. Here, we report that circulating cell-free DNAs from plasma and serum of patients with MDS can be used to detect genetic/epigenetic abnormalities. The plasma DNA concentration was found to be relatively high in patients with higher blast cell counts in BM, and accumulation of DNA fragments from mono-/di-nucleosomes was confirmed. Using serial peripheral blood (PB) samples from patients treated with hypomethylating agents, global methylation analysis using bisulfite pyrosequencing was performed at the specific CpG sites of the LINE-1 promoter. The results confirmed a decrease of the methylation percentage after treatment with azacitidine (days 3-9) using DNAs from plasma, serum, and PB mono-nuclear cells (PBMNC). Plasma DNA tends to show more rapid change at days 3 and 6 compared with serum DNA and PBMNC. Furthermore, the TET2 gene mutation in DNAs from plasma, serum, and BM cells was quantitated by pyrosequencing analysis. The existence ratio of mutated genes in plasma and serum DNA showed almost equivalent level with that in the CD34+/38- stem cell population in BM. These data suggest that genetic

  6. PTEN and p16 genes as epigenetic biomarkers in oral squamous cell carcinoma (OSCC): a study on south Indian population.

    Science.gov (United States)

    Sushma, P S; Jamil, Kaiser; Kumar, P Uday; Satyanarayana, U; Ramakrishna, M; Triveni, B

    2016-06-01

    Phosphatase and tensin homolog (PTEN) and p16INK4a (p16) genes are tumor suppressor genes, associated with epigenetic alterations. PTEN and p16 promoter hypermethylation is a major epigenetic silencing mechanism leading to cancer. The cooperation between PTEN and p16 in pathogenesis of cancers suggest that their combination might be considered as potential molecular marker for specific subgroups of patients. Hence, the present study aimed to investigate whether PTEN and p16 promoter methylations were involved in oral squamous cell carcinoma (OSCC) in south Indian subjects. DNA methylation quantitative analyses of the two candidate tumor suppressor genes PTEN and p16 were performed by methylation-specific polymerase chain reaction (MSP). Fifty OSCC biopsy samples and their corresponding non-malignant portions as controls were studied comparatively. The methylation status was correlated with the clinical manifestations. Twelve out of 50 patients (24 %) were found to be methylated for PTEN gene, whereas methylation of the p16 gene occurred in 19 out of 50 cases (38 %). A statistically significant result was obtained (P = p16 genes. PTEN and p16 promoter methylation may be the main mechanism leading to the low expression of PTEN and p16 genes indicating the progress of tumor development. Our data suggest that a low PTEN and p16 expression due to methylation may contribute to the cancer progression and could be useful for prognosis of OSCC. Therefore, analysis of promoter methylation in such genes may provide a biomarker valuable for early detection of oral cancer.

  7. A novel method to quantify local CpG methylation density by regional methylation elongation assay on microarray

    Directory of Open Access Journals (Sweden)

    Qiao Yingjuan

    2008-01-01

    Full Text Available Abstract Background DNA methylation based techniques are important tools in both clinical diagnostics and therapeutics. But most of these methods only analyze a few CpG sites in a target region. Indeed, difference of site-specific methylation may also lead to a change of methylation density in many cases, and it has been found that the density of methylation is more important than methylation of single CpG site for gene silencing. Results We have developed a novel approach for quantitative analysis of CpG methylation density on the basis of microarray-based hybridization and incorporation of Cy5-dCTP into the Cy3 labeled target DNA by using Taq DNA Polymerase on microarray. The quantification is achieved by measuring Cy5/Cy3 signal ratio which is proportional to methylation density. This methylation-sensitive technique, termed RMEAM (regional methylation elongation assay on microarray, provides several advantages over existing methods used for methylation analysis. It can determine an exact methylation density of the given region, and has potential of high throughput. We demonstrate a use of this method in determining the methylation density of the promoter region of the tumor-related gene MLH1, TERT and MGMT in colorectal carcinoma patients. Conclusion This technique allows for quantitative analysis of regional methylation density, which is the representative of all allelic methylation patterns in the sample. The results show that this technique has the characteristics of simplicity, rapidness, specificity and high-throughput.

  8. A COMPARATIVE ANALYSIS OF METHYLATION STATUS OF TUMOR SUPPRESSOR GENES IN PAIRED BIOPSY AND SERUM SAMPLES FROM CERVICAL CANCER PATIENTS AMONG NORTH INDIAN POPULATION.

    Science.gov (United States)

    Jha, A K; Sharma, V; Nikbakht, M; Jain, V; Sehgal, A; Capalash, N; Kaur, J

    2016-02-01

    Tumor-specific genetic or epigenetic alterations have been detected in serum DNA in case of various types of cancers. In breast cancer, the detection of tumor suppressor gene hypermethylation has been reported in several body fluids. Promoter hypermethylation of some genes like MYOD1, CALCA, hTERT etc. has also been detected in serum samples from cervical cancer. The present study is the first report on the comparison of promoter hypermethylation of tumor suppressor genes likep14, p15, p16, p21, p27, p57, p53, p73, RARβ2, FHIT, DAPK, STAT1 and-RB1 genes in paired biopsy and serum samples from cervical cancer patients among north Indian population. This is also the first report on the hypermethylation of these genes in serum samples from cervical cancer patients among north Indian population. According to the results of the present study, promoter hypermethylation of these genes can also be detected in serum samples of cervical cancer patients. The sensitivity of detection of promoter hypermethylation in serum samples of cervical cancer patients as compared to paired biopsy samples was found to be around 83.3%. It was observed that promoter hypermethylation was mainly observed in the serum samples in the higher stages and very rarely in the lower stages. The present study clearly showed that serum of patients with cervical cancer can also be used to study methylated genes as biomarkers.

  9. DETECTION OF TELOMERASE ACTIVITY IN BREAST CARCINOMA

    Institute of Scientific and Technical Information of China (English)

    Yang Wentao; Xu Liangzhong; Zhang Taiming; Zhu weiping; Li Xiaomei; Jin Aiping

    1998-01-01

    Objective:To investigate the significance of telomerase activity in breast carcinoma with its respect to axillary lymph node status. Methods: Telomerase activity was analyzed in 88 breast carcinomas and 16benign breast lesions, using polymerase chain reaction (PCR)-based telomeric repeat amplification protocol (TRAP) assay. Results: Telomerase activity was detected in 75 (85%) of 88 breast carcinomas (including three breast carcinomas in situ which were all positive for telomerase activity), whereas in benign breast lesions analyzed only 2(12.5%) of 16 cases were positive for telomerase activity. The difference between the two groups was statistically significant (P<0.001). Besides,telomerase activity was expressed significantly higher in node-positive breast carcinoma (93%) than in nodenegative ones (77%) (P<0.05). Conclusion: Our results suggest that telomerase activation plays an important role during breast carcinoma development. It is possible that this enzyme may serve as an early indication of breast carcinoma.

  10. Present Status of Diagnosis and Surgical Treatment in Papillary Thyroid Carcinoma%甲状腺乳头状癌外科诊治现状

    Institute of Scientific and Technical Information of China (English)

    嵇庆海

    2001-01-01

    The article described the diagnostic and therapeutic methods of papillary thyroid carcinoma at present, and put forward their own opinions.%文章主要叙述了目前甲状腺乳头状癌的各种诊断方法和治疗措施,并对此提出了相应的看法与认识。

  11. Immunoexpression status and prognostic value of mammalian target of rapamycin and hypoxia-induced pathway members in papillary cell renal cell carcinomas.

    Science.gov (United States)

    Chaux, Alcides; Schultz, Luciana; Albadine, Roula; Hicks, Jessica; Kim, Jenny J; Allaf, Mohamad E; Carducci, Michael A; Rodriguez, Ronald; Hammers, Hans-Joerg; Argani, Pedram; Reuter, Victor E; Netto, George J

    2012-12-01

    Dysregulation of the mammalian target of rapamycin and hypoxia-induced pathways has been consistently identified in clear cell renal cell carcinomas. However, experience with non-clear cell renal cell carcinoma subtypes is scant. In this study, we evaluated the immunohistochemical expression of upstream (PTEN and phosphorylated AKT) and downstream (phosphorylated S6 and 4EBP1) effectors of the mammalian target of rapamycin pathway, as well as related cell-cycle proteins (p27 and c-MYC), and a member of the hypoxia-induced pathway (HIF-1α) in 54 patients with papillary renal cell carcinoma treated by nephrectomy. PTEN was lower in tumor than in normal kidney, and loss of PTEN expression was found in 48% of the patients. In tumor tissues, phosphorylated S6, 4EBP1, and HIF-1α were higher than in normal kidney. Conversely, scores of p27 were lower in tumor than in normal kidney. Finally, scores of c-MYC and phosphorylated AKT were similar in tumor and in normal kidney. Overall mortality and cancer-specific mortality were 24% and 11%, respectively. Tumor progression was observed in 17% of the patients. None of the tested biomarkers predicted cancer-specific mortality or tumor progression. As expected, patients with high T-stage tumors had higher hazard ratios for cancer-specific mortality (hazard ratio, 6.9) and tumor progression (hazard ratio, 6.7). Patients with higher Fuhrman grades also had higher risks for cancer-specific mortality (hazard ratio, 11.4) and tumor progression (hazard ratio, 4.5). In summary, our study provides evidence of dysregulation of the mammalian target of rapamycin and hypoxia-induced pathways in papillary renal cell carcinoma. Immunohistochemistry for members of the mammalian target of rapamycin pathway and for HIF-1α lacked prognostic significance in our cohort.

  12. 视网膜母细胞瘤中MGMT启动子区甲基化状态及其产物表达%The MGMT promoter methylation status and its expression level in retinoblastoma

    Institute of Scientific and Technical Information of China (English)

    王怡琛; 李彬; 赵博文; 张浩; 高飞; 张志豹

    2012-01-01

    Objective To detect the methylation status of the promoter of O6 methylguanine DNA meghyltransferase {MGMT) gene and the expression level of MCMT mRNA and protein. Design Experimental Study. Participants 20 paraffin embedded RB tissues, 4 RB cell lines (Y79, SO-Rb50, SO-Rb50/VCR, WERI-RB1). Methods A series of 20 paraffin embedded RB tissue samples and 4 RB cell line were subjected to mehtylation-specific PCR (MSP) analysis to evaluate the methylation status of the MGMT promoter. Further, the expression of MGMT mRNA and protein were studied by Realtime PCR and Western Blot. The clinical data (e.g. Gender, age of onset) and pathological features (e.g. The realm of tumor encroachment, neovascularization in iris, NVI) were also collected. Main Outcome Measures Methylation status of the MGMT promoter, mRNA and protein expression level of MCMT, data of clinical and pathological features. Results Among the 20 paraffin embedded RB tissue samples, 12 cases (60%) were partially/completely methylated, the remaining 8 cases (40%) showed MGMT promoter unmethylated. As to the 4 RB cell lines, Y79, SO-Rb50, SO-Rb50/VCR were partially methylated, while WERI-RB1 was unmethylated. MGMT mRNA and protein were expressed in all 4 RB cell lines. The mRNA expression level of WERI-RBK 1.000±0.040) was higher than the other 3 cell lines( Y79,0.617±0.026;SO-Rb50,0.356±0.020 ;SO-Rb50/VCR, 0.389±0.017), the differences among which were statistically significant (P0.05). Conclusions There was a comparatively high MGMT promoter methylation rate in RB pathologic samples and cell lines, which had a corresponding effect to the expression level of MGMT mRNA and pro-tein. (Ophthalmol CHN, 2012, 21:121-126)%目的 检测视网膜母细胞瘤(RB)中,06-甲基鸟嘌呤DNA甲基转移酶(MGMT)基因启动子区甲基化状态及mRNA、蛋白产物的表达情况,并探讨其与RB临床、组织病理学特征的可能关系.设计 实验研究.研究对象 石蜡包埋RB组织20例,RB细胞系4

  13. Methylation Linear Discriminant Analysis (MLDA for identifying differentially methylated CpG islands

    Directory of Open Access Journals (Sweden)

    Vass J Keith

    2008-08-01

    Full Text Available Abstract Background Hypermethylation of promoter CpG islands is strongly correlated to transcriptional gene silencing and epigenetic maintenance of the silenced state. As well as its role in tumor development, CpG island methylation contributes to the acquisition of resistance to chemotherapy. Differential Methylation Hybridisation (DMH is one technique used for genome-wide DNA methylation analysis. The study of such microarray data sets should ideally account for the specific biological features of DNA methylation and the non-symmetrical distribution of the ratios of unmethylated and methylated sequences hybridised on the array. We have therefore developed a novel algorithm tailored to this type of data, Methylation Linear Discriminant Analysis (MLDA. Results MLDA was programmed in R (version 2.7.0 and the package is available at CRAN 1. This approach utilizes linear regression models of non-normalised hybridisation data to define methylation status. Log-transformed signal intensities of unmethylated controls on the microarray are used as a reference. The signal intensities of DNA samples digested with methylation sensitive restriction enzymes and mock digested are then transformed to the likelihood of a locus being methylated using this reference. We tested the ability of MLDA to identify loci differentially methylated as analysed by DMH between cisplatin sensitive and resistant ovarian cancer cell lines. MLDA identified 115 differentially methylated loci and 23 out of 26 of these loci have been independently validated by Methylation Specific PCR and/or bisulphite pyrosequencing. Conclusion MLDA has advantages for analyzing methylation data from CpG island microarrays, since there is a clear rational for the definition of methylation status, it uses DMH data without between-group normalisation and is less influenced by cross-hybridisation of loci. The MLDA algorithm successfully identified differentially methylated loci between two classes of

  14. Effects of 5-Aza-dC and TSA on methylation and expression of 3-OST-2 gene in human hepatocelluar carcinoma cells%5-Aza-dC和TSA对肝细胞癌细胞株中3-OST-2甲基化和基因表达的影响

    Institute of Scientific and Technical Information of China (English)

    姚树哲; 陈海燕; 张晓莹; 盛燕; 周庚寅; 张翠娟

    2013-01-01

    Purpose To investigate the effects of 5 -Aza-dC and TSA on promoter methylation and gene expression of 3 -OST-2 as well as the expression of nuclear transcription factor UHRF1. Methods The changes of promoter methylation of 3-OST-2 hefore and after the treatment of 5-Aza-dC and/or TSA were detected hy methylation specific PCR ( MSP ). The changes of mRNA and protein expression of 3-OST-2 or UHRF1 were determined hy real time RT-PCR and immunocytochemistry, respectively. Results 3-OST-2 was completely methylated in hepatocellular carcinoma cells and was partially reversed hy 5-Aza-dC or TSA alone, consistently, mRNA was increased hy 2. 7-fold and 4. 9-fold, respectively. After the treatment of the inhihitors in comhination, 3-OST-2 methylation was completely reversed and mRNA was significantly increased by 9. 1-fold. UHRF1 mRNA and protein expression were both inhibited by 5-Aza-dC or TSA, and TSA was more efficient than 5-Aza-dC ( P <0. 01 ). There was synergistic effect between the two inhibitors( P < 0. 01 ). Conclusions Promoter methylation and histone modifications both cause the decrease of 3-OST-2 expression. Either 5-Aza-dC or TSA can reverse 3-OST-2 methylation and increase its expression, and the inhibition of UHRF1 may be one of the mechanisms.%目的 探讨5-Aza-dC和TSA对肝细胞癌(hepatocellular carcinoma,HCC)细胞中3-OST-2甲基化水平、基因表达以及对核转录因子UHRF1(ubiquitin-like with PHD and ring finger domains 1,也称为ICBP90/NP95)表达的影响.方法 采用甲基化特异性PCR(methylation specific PCR,MSP)法检测药物作用前后HCC细胞株中3-OST-2甲基化的状态改变;采用实时荧光定量RT-PCR法检测3-OST-2和UHRF1 mRNA的表达变化;采用细胞爬片免疫组化法检测UHRF1蛋白表达.结果 3-OST-2在HCC细胞株中呈完全甲基化状态,5-Aza-dC或TSA均能部分逆转3-OST-2甲基化,其mRNA表达分别增加2.7倍和4.9倍,两种药物联合处理后,3-OST-2甲基化被

  15. Squamous Cell Carcinoma

    Science.gov (United States)

    ... Kids’ zone Video library Find a dermatologist Squamous cell carcinoma Overview Squamous cell carcinoma: This man's skin ... a squamous cell carcinoma on his face. Squamous cell carcinoma: Overview Squamous cell carcinoma (SCC) is a ...

  16. Assessing p16 Status of Oropharyngeal Squamous Cell Carcinoma by Combined Assessment of the Number of Cells Stained and the Confluence of p16 Staining: A Validation by Clinical Outcomes.

    Science.gov (United States)

    Barasch, Samuel; Mohindra, Pranshu; Hennrick, Kenneth; Hartig, Gregory K; Harari, Paul M; Yang, David T

    2016-09-01

    Human papillomavirus-related oropharyngeal squamous cell carcinoma (OPSCC) has favorable prognosis relative to other head and neck squamous cell carcinomas. Criteria for predicting human papillomavirus status based upon p16 staining, including difficult cases with partial staining patterns, have been developed; however, clinical validation of these criteria and the clinical significance of partial p16 staining have not been reported. Eighty-one archival OPSCC cases were initially stained for p16 by immunohistochemistry with clone G175-405. The percentage of p16 cells and percentage of confluence of p16 cells were categorized as 25%, 26% to 75%, or >75%. Of all cases, 16 (20%) had partial p16 expression, with 26% to 75% p16 cells. Applying previously developed criteria of >75% p16 cells or >50% positive cells with >25% confluence, 48 (59%) patients were categorized p16 and demonstrated expected clinical characteristics and superior disease-free survival and overall survival (Pp16 patients. By themselves, the partial staining patients had intermediate outcomes; however, separating the partial staining cases by degree of confluence showed that those with >75% confluence had superior disease-free survival (P=0.042). When the 16 original partial staining cases were re-stained with the alternative anti-p16 E6H4 clone, p16 status remained concordant for all cases, but only 3 of the 16 were interpreted as demonstrating partial staining. This report shows that the prevalence of partial p16 staining varies with the antibody utilized and clinically validates the application of a graded evaluation of both the number as well as confluence of positive cells for risk stratification of patients with OPSCC.

  17. Expression of ATP7B in human gastric cardiac carcinomas in comparison with distal gastric carcinomas

    Institute of Scientific and Technical Information of China (English)

    Da-Long Wu; Hui-Xing Yi; Feng-Ying Sui; Xiao-Hong Jiang; Xiao-Ming Jiang; Ying-Ying Zhao

    2006-01-01

    AIM: To analyze expression of ATP7B in gastric cardiac adenocarcinomas, its clinicopathologic significance, in comparison with distal gastric adenocarcinomas.METHODS: Immunohistochemical avidin-biotin peroxidase complex method was applied to detect the expression of ATP7B in 49 cases of cardiac carcinomas,the corresponding adjacent non-neoplastic epithelium and 55 cases of distal gastric carcinomas.RESULTS: The proportion of ATP7B positive samples in gastric cardiac carcinomas (51.0%, 25 of 49) was significantly higher than that in the corresponding adjacent non-neoplastic epithelium (22.4%, 11 of 49)(P = 0.003). ATP7B expression in poorly differentiated gastric cardiac carcinomas was significantly higher than that in well/moderately differentiated gastric cardiac carcinomas (P = 0.030). ATP7B expression in gastric cardiac carcinomas was independent of age, tumor size, nodal stage and metastasis status. ATP7B protein was detected in 30.9% (17/55 cases) of distal gastric carcinomas, markedly lower than that in gastric cardiac carcinomas (P = 0.037).CONCLUSION: ATP7B protein is frequently overexpressed in gastric cardiac carcinomas, and correlated with the differentiation of cardiac carcinoma. ATP7B expression in gastric cardiac carcinomas is significantly higher than that in distal gastric carcinomas, which might partially explain the difference of chemotherapy response and prognosis between these two gastric carcinomas.

  18. Methyl gallate

    Directory of Open Access Journals (Sweden)

    Silvina Pagola

    2009-02-01

    Full Text Available The crystal structure of the title compound (systematic name: methyl 3,4,5-trihydroxybenzoate, C8H8O5, is composed of essentially planar molecules [maximum departures from the mean carbon and oxygen skeleton plane of 0.0348 (10 Å]. The H atoms of the three hydroxyl groups, which function as hydrogen-bond donors and acceptors simultaneously, are oriented in the same direction around the aromatic ring. In addition to two intramolecular hydrogen bonds, each molecule is hydrogen bonded to six others, creating a three-dimensional hydrogen-bonded network.

  19. Anti-tumor effect of 5-aza-2'-deoxycytidine by inhibiting telomerase activity in hepatocellular carcinoma cells

    Institute of Scientific and Technical Information of China (English)

    Shuang-Fen Tao; Chang-Song Zhang; Xian-Ling Guo; Yun Xu; Shan-Shan Zhang; Jian-Rui Song; Rong Li

    2012-01-01

    AIM:To investigate the effect of the demethylating reagent 5-aza-2'-deoxycitidine (DAC) on telomerase activity in hepatocellular carcinoma (HCC) cell lines,SMMC-7721 and HepG2.METHODS:The related gene expression in cell lines was examined by real-time reverse transcription-polymerase chain reaction and Western blotting analysis.The telomerase activity was examined by telomeric repeat amplification protocol-enzyme-linked immunosorbent assay and DNA methylation was determined by methylation-specific polymerase chain reaction.RESULTS:The telomerase activity was significantly reduced in both cell lines treated with DAC,accompanied by downregulation of telomerase reverse transcriptase (hTERT).We also observed the effect of DAC on the methylation status of hTERT promoter and the expression of regulatory genes,such as c-myc,p15,p16,p21,E2F1,and WT1.The methylation status of hTERT promoter could be reversed in SMMC-7721 by DAC,but not in HepG2 cells.However,p16 expression could be reactivated by demethylation of its promoter,and c-Myc expression was repressed in both cell lines.Moreover,DAC could enhance the sensitivity to the chemotherapeutic agents,such as cisplatin,by induction of apoptosis of HCC cells.CONCLUSION:The DAC exerts its anti-tumor effects in HCC cells by inhibiting the telomerase activity.

  20. 首次住院治疗鼻咽癌患者心理健康状况调查分析%Analysis of the first hospitalization for mental health status of patients with nasopharyngeal carci-noma investigation

    Institute of Scientific and Technical Information of China (English)

    彭俊华; 张小玉; 陈楚君; 陈桂玲

    2014-01-01

    目的:对首次住院治疗鼻咽癌患者的心理健康状况进行调查分析,为今后实施有效的护理干预提供理论依据。方法采用SCL-90症状自评量表对150例首次住院治疗鼻咽癌患者进行问卷调查,与国内常模比较,分析其心理健康状况。结果首次住院治疗鼻咽癌患者SCL-90量表总均分、躯体化、抑郁、焦虑、精神病性分值显著高于常模组(P﹤0.05或0.01);人际关系和偏执两因子分显著低于常模组(P﹤0.05),此外强迫和敌对因子与我国常模比较差异无统计学意义( P﹥0.05)。结论首次住院治疗鼻咽癌患者的心理健康状况较正常人低,其存在比较广泛的心理健康问题,因此在治疗癌症的同时,应积极进行心理干预。%Objective To investigate and analyze the mental health status of first hospital treat-ment of patients with nasopharyngeal carcinoma,and provide a theoretical basis for the implementation of effective nursing intervention in the future. Methods One hundred and fifty cases of nasopharyngeal carcinoma who were first hospitalization were investigated and analyzed by SCL-90 symptom checklist, their psychological health status were analyzed compared with the national norm. Results Arong the first hospitalized patients with nasopharyngeal carcinoma,the SCL-90 scale total score,somatization,depres-sion,anxiety,psychopathy scores were significantly higher than that of the normal group( P﹤0. 05 or 0 . 01 );interpersonal relationship and paranoid two factor were significantly lower than that of the normal group( P﹤0 . 05 ),and the factors of force and hostility had no significant difference compared with norm in our country( P﹥0 . 05 ). Conclusions The status of psychological health of the first hospitalized naso-pharyngeal carcinoma is lower than that of the healthy people,they always had mental health problems, so we should actively carry out psychological intervention based on the

  1. Glucose utilization in the brain during acute seizure is a useful biomarker for the evaluation of anticonvulsants: effect of methyl ethyl ketone in lithium-pilocarpine status epilepticus rats

    Energy Technology Data Exchange (ETDEWEB)

    Yamada, Akifumi [Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka, 565-0871 (Japan)], E-mail: yamaaki@sahs.med.osaka-u.ac.jp; Momosaki, Sotaro; Hosoi, Rie [Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka, 565-0871 (Japan); Abe, Kohji [Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka, 565-0871 (Japan); Developmental Research Laboratories, Shionogi and Co., Ltd., Toyonaka, Osaka, 561-0825 (Japan); Yamaguchi, Masatoshi [Faculty of Pharmaceutical Sciences, Fukuoka University, Johnan, Fukuoka 814-0180 (Japan); Inoue, Osamu [Division of Health Sciences, Graduate School of Medicine, Osaka University, Osaka, 565-0871 (Japan)

    2009-11-15

    Enhancement of glucose utilization in the brain has been well known during acute seizure in various kinds of animal model of epilepsy. This enhancement of glucose utilization might be related to neural damage in these animal models. Recently, we found that methyl ethyl ketone (MEK) had both anticonvulsive and neuroprotective effects in lithium-pilocapine (Li-pilo) status epilepticus (SE) rat. In this article, we measured the uptake of [{sup 14}C]2-deoxyglucose ([{sup 14}C]DG) in the Li-pilo SE and Li-pilo SE with MEK rat brain in order to assess whether the glucose utilization was a useful biomarker for the detection of efficacy of anticonvulsive compounds. Significant increase of [{sup 14}C]DG uptake (45 min after the injection) in the cerebral cortex, hippocampus, amygdala and thalamus during acute seizure induced by Li-pilo were observed. On the other hand, the initial uptake of [{sup 14}C]DG (1 min after the injection) in the Li-pilo SE rats was not different from the control rats. Therefore, the enhancement of glucose metabolism during acute seizure was due to the facilitation of the rate of phosphorylation process of [{sup 14}C]DG in the brain. Pretreatment with MEK (8 mmol/kg) completely abolished the enhancement of glucose utilization in the Li-pilo SE rats. The present results indicated that glucose utilization in the brain during acute seizure might be a useful biomarker for the evaluation of efficacy of anticonvulsive compounds.

  2. Imaging findings of common benign renal tumors in the era of small renal masses: Differential diagnosis from small renal cell carcinoma: Current status and future perspectives

    Energy Technology Data Exchange (ETDEWEB)

    Woo, Sung Min; Cho, Jeong Yeon [Dept. of Radiology, Seoul National University College of Medicine, Seoul (Korea, Republic of)

    2015-02-15

    The prevalence of small renal masses (SRM) has risen, paralleling the increased usage of cross-sectional imaging. A large proportion of these SRMs are not malignant, and do not require invasive treatment such as nephrectomy. Therefore, differentation between early renal cell carcinoma (RCC) and benign SRM is critical to achieve proper management. This article reviews the radiological features of benign SRMs, with focus on two of the most common benign entities, angiomyolipoma and oncocytoma, in terms of their common imaging findings and differential features from RCC. Furthermore, the role of percutaneous biopsy is discussed as imaging is yet imperfect, therefore necessitating biopsy in certain circumstances to confirm the benignity of SRMs.

  3. Effects of methylation status of CpG islands on results of luciferase reporter assay%细胞荧光素酶报告基因试验受CpG岛甲基化状态影响

    Institute of Scientific and Technical Information of China (English)

    张宝珍; 邓大君

    2009-01-01

    Objective To investigate the effects of methylation status of CpG islands of endogenous E-cadherin (CDH1) gene on the promoter activity of corresponding genes in reporter assays. Methods The methylation statuses of CpG island of CDHI in 8 different cell lines were detected by methylation-specific PCR. CDH1 protein was analyzed by Western blotting. Two sets of pGL3 reporter vectors with different genotypes/haplotypes of the CDH1 promoter were constructed [pGI3-A(-73)/-C(-73)pGL3-H1/-H4]and used to transfect these cell lines. The differences between these promoter reporter vectors were analyzed by t-test. Results (1) CDH1 CpG island was unmethylated in AGS, MCF7, MKN74, and PC-3 cell lines,expressed in MCF7, MKN74, and PC-3 ,but not in AGS. Expression of CDH1 was silenced by methylation in HeLa, BGC823, A549, and RKO cell lines. (2) In the four CDH1 -unmethylated MCFT, M KN74, PC-3, and AGS cell lines ,the promoter activities of pGI3-C(-73)(as 0. 78±0. 10,0. 17±0.01,0. 11±0. 01,1.19±0. 18)were significantly higher than those of pGL3-A(-73)(as 0. 30±0. 08,0. 07±0. 01,0. 07±0. 01,0. 39±0. 04) (t values are -6. 298, -12. 349, -8. 128, -7.388, and P<0. 01). However, in the four C DH1 -methylated HeLa, BGC823, A549, and RKO cell lines, the promoter activity of pGL3-C(-73)(as 0. 09±0. 02,0. 13±0. 02,0. 05±0. 01,0. 01±0. 00) was significantly lower than that of pGL3-A(-73)(as 0. 16±0. 01,0.25±0.01,0. 11±0.03,0.03±0.00) (t valued at 5.958,11. 189,3. 661,13. 866,and P<0.05). (3) In the unmethylated MKN74 and methylated RKO cell lines, the promoter activities of pGI3-H1/-H4 were obviously and contrarily different(as 1.57±0. 23/0. 94±0. 06 and 0. 38±0. 02/0. 50±0. 04 ,t values were 4. 577 and -4. 915 ,P values were 0. 010 and 0. 003). Conclusion The methylation status of CpG island of the target gene in the tested cell lines affects the promoter activity in Reporter Assay significantly. The most active one may be the most suppressive one.%目的 研究细

  4. Sebaceous Carcinoma

    Science.gov (United States)

    ... Hill Medical; 2008. p. 240-9. Nelson BR, Hamlet KR, Gillard M et al. “Sebaceous carcinoma.” J ... Acad Dermatol . 2003 48:401-8. Nelson BR, Hamlet KR, Gillard M et al . “Sebaceous carcinoma.” J ...

  5. Expression of the p16(INK4a) gene product, methylation of the p16(INK4a) promoter region and expression of the polycomb-group gene BMI-1 in squamous cell lung carcinoma and premalignant endobronchial lesions.

    NARCIS (Netherlands)

    Breuer, R.H.J.; Snijders, P.J.F.; Sutedja, T.G.; Sewalt, R.G.A.B.; Otte, A.P.; Postmus, P.E.; Meijer, C.J.L.M.; Raaphorst, F.M.; Smit, E.F.

    2005-01-01

    It is generally assumed that squamous cell carcinoma develops in a stepwise manner from normal bronchial epithelium towards cancer by the accumulation of (epi)genetic alterations. Several mechanisms including mutations and homozygous deletions or hypermethylation of the p16(INK4a) promoter region ca

  6. Genetic and Epigenetic Alterations of DLC-1, a Candidate Tumor Suppressor Gene, in Nasopharyngeal Carcinoma

    Institute of Scientific and Technical Information of China (English)

    Dan PENG; Cai-Ping REN; Hong-Mei YI; Liang ZHOU; Xu-Yu YANG; Hui LI; Kai-Tai YAO

    2006-01-01

    The DLC-1 gene, located at the human chromosome region 8p22, behaves like a tumor suppressor gene and is frequently deleted in diverse tumors. The deletion of 8p22 is not an uncommon event in nasopharyngeal carcinoma (NPC), therefore we explored the expression levels of the DLC-1 gene in NPCs and NPC cell lines by reverse transcription-polymerase chain reaction. The results showed the mRNA level of DLC-1 was downregulated. To identify the mechanism of DLC-1 downregulation in NPC, we investigated the methylation status of the DLC-1 gene using methylation-specific PCR, and found that 79% (31 of 39) of the NPC tissues and two DLC-1 nonexpressing NPC cell lines, 6-10B and 5-8F, were methylated in the DLC-1 CpG island. Microsatellite PCR was also carried out, and loss of heterozygosity was found at four microsatellite sites (D8S552, D8S1754, D8S1790 and D8S549) covering the whole DLC-1 gene with ratios of 33% (4 of 12 informative cases), 18% (2 of 11), 5% (1 of 18), and 25% (3 of 12), respectively. Taken together, our results suggest that DLC-1 might be an NPC-related tumor suppressor gene affected by aberrant promoter methylation and gene deletion.

  7. 血浆Ras相关区域家族蛋白1A基因甲基化在肝细胞癌分子诊断中的价值%Clinical significance of methylation of plasma Ras-association domain family 1A gene in the molecular diagnosis of hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    费伯健; 黄朝晖; 华东; 胡瑜; 程之红; 余坚

    2011-01-01

    Objective: To establish a methylation-sensitive restriction enzymes-based quantitative PCR (MSRE-qPCR) method for methylation analysis of Ras-association domain family 1A (RASSF1A) gene, and to further assess the clinical value of plasma methylation analysis using this method for noninvasive diagnosis of hepatocellutar carcinoma (HCC). Methods: MRSE Hha I was used to digest genomic DNA samples, and the digestion result was evaluated by using qPCR. Then the MSRE-qPCR method for methylation analysis of RASSF\\A gene was established. The efficacy of this method was tested in 45 liver tissues (20 surgically resected HCC specimens and the matched non-cancerous tissues, as well as 5 normal liver tissues), and which was further validated by using bisulfite sequencing PCR (BSP). The results of MRSE-qPCR were compared with those of methylation-specific PCR (MSP) assay. The established MSRE-qPCR method was used to detect the RASSF]A methylation levels in 150 plasma samples from 72 cases of HCC, 37 cases of benign liver diseases such as liver cirrhosis and chronic hepatitis, and 41 normal controls, and the associations of RASSF1A methylation level with the clinicopathological parameters of patients with HCC were evaluated.Results: The established MSRE-qPCR method could detect as low as 1% methylated target sites in given DNA samples. Of the 20 HCC tissues, 14 (70%) were hypermethylated in the target CpC of RASSF]A promoter. Five matched non-cancerous tissues were also found to be methylated, whereas no methylated RASSF]A sequence was detected in 5 normal liver tissues. The result of MSRE-qPCR was accurate and comparable with that of MSP. The positive rate of RASSF\\A methylation in plasma samples from patients with HCC was 65.3% (47/72), which was significantly higher than those from patients with benign liver diseases (3/37, 8.1%) and normal controls (1/41, 2.4%) (f<0.000 1). Combined analysis of plasma RASSF\\A methylation and serum AFP revealed an increased diagnostic

  8. Plasma DNA tissue mapping by genome-wide methylation sequencing for noninvasive prenatal, cancer, and transplantation assessments

    Science.gov (United States)

    Sun, Kun; Jiang, Peiyong; Chan, K. C. Allen; Wong, John; Cheng, Yvonne K. Y.; Liang, Raymond H. S.; Chan, Wai-kong; Ma, Edmond S. K.; Chan, Stephen L.; Cheng, Suk Hang; Chan, Rebecca W. Y.; Tong, Yu K.; Ng, Simon S. M.; Wong, Raymond S. M.; Hui, David S. C.; Leung, Tse Ngong; Leung, Tak Y.; Lai, Paul B. S.; Chiu, Rossa W. K.; Lo, Yuk Ming Dennis

    2015-01-01

    Plasma consists of DNA released from multiple tissues within the body. Using genome-wide bisulfite sequencing of plasma DNA and deconvolution of the sequencing data with reference to methylation profiles of different tissues, we developed a general approach for studying the major tissue contributors to the circulating DNA pool. We tested this method in pregnant women, patients with hepatocellular carcinoma, and subjects following bone marrow and liver transplantation. In most subjects, white blood cells were the predominant contributors to the circulating DNA pool. The placental contributions in the plasma of pregnant women correlated with the proportional contributions as revealed by fetal-specific genetic markers. The graft-derived contributions to the plasma in the transplant recipients correlated with those determined using donor-specific genetic markers. Patients with hepatocellular carcinoma showed elevated plasma DNA contributions from the liver, which correlated with measurements made using tumor-associated copy number aberrations. In hepatocellular carcinoma patients and in pregnant women exhibiting copy number aberrations in plasma, comparison of methylation deconvolution results using genomic regions with different copy number status pinpointed the tissue type responsible for the aberrations. In a pregnant woman diagnosed as having follicular lymphoma during pregnancy, methylation deconvolution indicated a grossly elevated contribution from B cells into the plasma DNA pool and localized B cells as the origin of the copy number aberrations observed in plasma. This method may serve as a powerful tool for assessing a wide range of physiological and pathological conditions based on the identification of perturbed proportional contributions of different tissues into plasma. PMID:26392541

  9. Arginine Methylation of SREBP1a via PRMT5 Promotes De Novo Lipogenesis and Tumor Growth.

    Science.gov (United States)

    Liu, Liu; Zhao, Xiaoping; Zhao, Li; Li, Jiajin; Yang, Hao; Zhu, Zongping; Liu, Jianjun; Huang, Gang

    2016-03-01

    Dysregulation of the sterol regulatory element-binding transcription factors sterol regulatory element-binding protein (SREBP) and SREBF activates de novo lipogenesis to high levels in cancer cells, a critical event in driving malignant growth. In this study, we identified an important posttranslational mechanism by which SREBP1a is regulated during metabolic reprogramming in cancer cells. Mass spectrometry revealed protein arginine methyltransferase 5 (PRMT5) as a binding partner of SREBP1a that symmetrically dimethylated it on R321, thereby promoting transcriptional activity. Furthermore, PRMT5-induced methylation prevented phosphorylation of SREBP1a on S430 by GSK3β, leading to its disassociation from Fbw7 (FBXW7) and its evasion from degradation through the ubiquitin-proteasome pathway. Consequently, methylation-stabilized SREBP1a increased de novo lipogenesis and accelerated the growth of cancer cells in vivo and in vitro. Clinically, R321 symmetric dimethylation status was associated with malignant progression of human hepatocellular carcinoma, where it served as an independent risk factor of poor prognosis. By showing how PRMT5-induced methylation of SREBP1a triggers hyperactivation of lipid biosynthesis, a key event in tumorigenesis, our findings suggest a new generalized strategy to selectively attack tumor metabolism.

  10. Up-regulation of expression and lack of 5' CpG island hypermethylation of p16 INK4a in HPV-positive cervical carcinomas

    Directory of Open Access Journals (Sweden)

    Frank Georgy A

    2007-03-01

    Full Text Available Abstract Background High risk type human papilloma viruses (HR-HPV induce carcinomas of the uterine cervix by expressing viral oncogenes E6 and E7. Oncogene E7 of HR-HPV disrupts the pRb/E2F interaction, which negatively regulates the S phase entry. Expression of tumor suppressor p16ink4a drastically increases in majority of HR-HPV associated carcinomas due to removal of pRb repression. The p16ink4a overexpression is an indicator of an aberrant expression of viral oncogenes and may serve as a marker for early diagnostic of cervical cancer. On the other hand, in 25–57% of cervical carcinomas hypermethylation of the p16 INK4a promoter has been demonstrated using a methylation-specific PCR, MSP. To evaluate a potential usage of the p16 INK4a 5' CpG island hypermethylation as an indicator of tumor cell along with p16ink4a overexpression, we analyzed the methylation status of p16 INK4a in cervical carcinomas Methods Methylation status of p16 INK4a was analyzed by MSP and by bisulfite-modified DNA sequencing. The expression of p16ink4a was analyzed by RT-PCR and by immunohistochemical technique. Results The extensive methylation within p16 INK4a 5' CpG island was not detected either in 13 primary cervical carcinomas or in 5 cancer cell lines by bisulfite-modified DNA sequencing (including those that were positive by MSP in our hands. The number and distribution of rare partially methylated CpG sites did not differ considerably in tumors and adjacent normal tissues. The levels of the p16 INK4a mRNA were increased in carcinomas compared to the normal tissues independently of the number of partially methylated CpGs within 5'CpG island. The transcriptional activation of p16 INK4a was accompanied by p16ink4a cytoplasmic immunoreactivity in the majority of tumor cells and presence of a varied number of the p16 positive nuclei in different tumors. Conclusion Hypermethylaion of the p16INK4a 5' CpG island is not a frequent event in HR-HPV-positive cervical

  11. Homogalacturonan Methyl-Esterification and Plant Development

    Institute of Scientific and Technical Information of China (English)

    Sebastian Wolf; Gregory Mouille; Jérome Pelloux

    2009-01-01

    The ability of a plant cell to expand is largely defined by the physical constraints imposed by its cell wall. Accordingly, cell wall properties have to be regulated during development. The pectic polysaccharide homogalacturonan is a major component of the plant primary walls. Biosynthesis and in muro modification of homogalacturonan have recently emerged as key determinants of plant development, controlling cell adhesion, organ development, and phyllo-tactic patterning. This review will focus on recent findings regarding impact of homogalacturonan content and methyl-esterification status of this polymer on plant life. De-methyl-esterification of homogalacturonan occurs through the action of the ubiquitous enzyme 'pectin methyl-esterase'. We here describe various strategies developed by the plant to finely tune the methyl-esterification status of homogalacturonan along key events of the plant lifecycle.

  12. Nutritional Status and Nutritional Intervention of Patients with Nasopharyngeal Carcinoma%鼻咽癌患者的营养状况及营养干预

    Institute of Scientific and Technical Information of China (English)

    韦燕

    2013-01-01

    鼻咽癌患者在放化疗治疗时营养不足和营养风险问题比较普遍,营养不良反过来影响患者体质与行为能力,限制患者对治疗的依从性和耐受性.对住院的鼻咽癌患者进行营养筛查,能早期识别和确定存在营养风险和营养不良的患者,从而给予及时有效的营养支持.营养干预的时间是个体化很强的问题,不同患者对治疗的耐受性不同也决定了干预时间的个体化.营养干预的方式、干预的时机及持续的时间仍需更多的研究来进一步明确.%Undernutrition and nutritional risk of nasopharyngeal carcinoma patients during radiotherapy and chemotherapy are common.Undernutrition affects the patient's physical and capacity,limits the compliance and tolerance of the treatment.Nutritional screening for hospitalized patients with nasopharyngeal carcinoma can early i-dentify and determine the existence of the nutritional risk and undernutrition, to give the effective nutrition support promptly.The timing of the nutrition intervention is highly individualized, different treatment tolerance of different patients determines the individualized intervention timing.Nutrition intervention methods, timing and the lasting time still need more research to further clarify.

  13. Cryotherapy for hepatocellular carcinoma

    DEFF Research Database (Denmark)

    Awad, Tahany; Thorlund, Kristian; Gluud, Christian

    2009-01-01

    BACKGROUND: Hepatocellular carcinoma is the most common primary malignant cancer of the liver. Evidence for the role of cryotherapy in the treatment of hepatocellular carcinoma is controversial. OBJECTIVES: The aim of this review is to evaluate the potential benefits and harms of cryotherapy...... status) comparing cryotherapy with or without co-intervention(s) to placebo, no treatment, or other control interventions were considered for the review. Due to the absence of randomised clinical trials, we searched for quasi-randomised studies as well as prospective cohort studies and retrospective...... for the assessment of benefit as the study results were stratified according to both the type of hepatic malignancy (primary or secondary) and the intervention group. This retrospective study compared percutaneous cryotherapy with percutaneous radiofrequency. The remaining studies were excluded for the analyses...

  14. Turning over DNA methylation in the mind

    Directory of Open Access Journals (Sweden)

    Ryan eLister

    2015-07-01

    Full Text Available Cytosine DNA methylation is a stable epigenetic modification with established roles in regulating transcription, imprinting, female X-chromosome inactivation, and silencing of transposons. Dynamic gain or loss of DNA methylation reshapes the genomic landscape of cells during early differentiation, and in post-mitotic mammalian brain cells these changes continue to accumulate throughout the phases of cortical maturation in childhood and adolescence. There is also evidence for dynamic changes in the methylation status of specific genomic loci during the encoding of new memories, and these epigenome dynamics could play a causal role in memory formation. However, the mechanisms that may dynamically regulate DNA methylation in neurons during memory formation and expression, and the function of such epigenomic changes in this context, are unclear. Here we discuss the possible roles of DNA methylation in encoding and retrieval of memory.

  15. Hypermethylation of Syk gene in promoter region associated with oncogenesis and metastasis of gastric carcinoma

    Institute of Scientific and Technical Information of China (English)

    Shui Wang; Yong-Bin Ding; Guo-Yu Chen; Jian-Guo Xia; Zhen-Yan Wu

    2004-01-01

    AIM: To investigate the rrelationship between methylation of Syk (spleen tyrosine kinase) gene in promoter region and oncogenesis, metastasis of gastric carcinoma. The relation between silencing of the Syk gene and methylation of Syk promoter region was also studied.METHODS: By using methylation-specific PCR (MSP)technique, the methylation of Syk promoter region in specimens from 61 gastric cancer patients (tumor tissues and adjacent normal tissues) was detected. Meanwhile, RTPCR was used to analyse syk expression exclusively.RESULTS: The expression of the Syk gene was detected in all normal gastric tissues. Syk expression in gastric carcinoma was lower in 14 out of 61 gastric cancer samples than in adjacent normal tissues (x2=72.3, P<0.05). No methylation of Syk promoter was found in adjacent normal tissues, hypermethylation of Syk gene in promoter was detected 21 cases in 61 gastric carcinoma patients. The rate of methylation of Syk promoter in gastric carcinoma was higher than that in adjacent normal tissues (x2=25.1,P<0.05). In 31 patients with lymph node metastasis, 17 were found with Syk promoter methylation. A significant difference was noted between two groups (x2=11.4, P<0.05).CONCLUSION: Hypermethylation leads to silencing of the Syk gene in human gastric carcinoma. Methylation of Syk promoter is correlated to oncogenesis and metastasis of gastric carcinoma. Syk is considered to be a potential tumor suppressor and anti-metastasis gene in human gastric cancer.

  16. Basal Cell Carcinoma

    Science.gov (United States)

    ... Kids’ zone Video library Find a dermatologist Basal cell carcinoma Overview Basal cell carcinoma: This skin cancer ... that has received years of sun exposure. Basal cell carcinoma: Overview Basal cell carcinoma (BCC) is the ...

  17. Effects of CDH1 gene promoter methylation on expression of E-cadherin and beta-catenin and its clinicopathological significance in colon carcinoma%上皮钙黏素1基因启动子甲基化对结肠癌上皮钙黏素和β-连接素表达的影响

    Institute of Scientific and Technical Information of China (English)

    李臣; 杨静; 董坚; 陈明清; 李文亮; 任俊宇; 陈圣雄; 李秋恬; 耿计伟; 缪延栋

    2011-01-01

    目的 探讨上皮钙黏素基因(CDH1)启动子甲基化与结肠癌上皮钙黏素(E-cadherin)及β-连接素(β-catenin)的表达及临床病理特征的关系.方法 采用甲基化特异性PCR技术检测68例结肠腺癌组织、癌旁组织及正常黏膜组织中CDH1基因启动子甲基化的状况.采用免疫组织化学法检测E-cadherin及β-catenin蛋白的表达.结果 癌旁组织及癌组织中CDH1启动子甲基化的阳性表达分别为32.4%(22/68)、57.4%(39/68),正常组织均为阴性表达(P<0.05).E-cadherin在正常组织、癌旁组织及腺癌组织中阳性表达率分别为92.6%、66.2%和44.1%.正常组织中β-catenin均表达于细胞膜上,无胞质和(或)胞核表达,而β-catenin在癌旁组织及癌组织中胞质和(或)胞核表达分别为29.4%和50.0%.CDH1基因启动子甲基化阳性率与E-cadherin表达则呈负相关(r=-0.312,P=0.01),与β-catenin胞质和(或)胞核表达呈正相关(r=0.309,P=0.018).CDH1基因启动子甲基化及E-cadherin、β-catenin的异常表达均与结肠癌分化程度及转移密切相关(P<0.05).结论 CDH1基因启动子甲基化可能是导致结肠癌E-cadherin与β-catenin异常表达及肿瘤侵袭性增强的重要原因.%Objective To investigate the relationship between methylation of the CDH1 gene promoter on the expression of E-cadherin and β-catenin, and to evaluate the correlation with clinicopathological characteristics of the colonic carcinoma. Methods Methylation specific PCR (MSP) was used to detect CDH1 gene promoter methylation in the cancer tissue, adjacent tissues and normal tissues in 68 patients. The expression of E-cadherin and β-catenin was determined by immunohistochemistry staining. Results The positive rate of CDH1 gene promoter methylation was 32.4% in adjacent tissues and 57.4% in cancer tissue, while no detectable methylation was found in all the normal tissues. The difference was statistically significant. The positive rate of E-cadherin was 92.6% in the

  18. 食管和胃双原发癌组织CDH1基因甲基化的表达及意义%The expression of CDH1 gene methylation in patients with esophagus and stomach double primary carcinoma and its significance

    Institute of Scientific and Technical Information of China (English)

    李永丽; 张立玮; 丁国瑾; 袁丽; 朱晓娅; 侯淑芸

    2011-01-01

    Objective To study the changes of CDH1 gene promoter CpG island methylation and its clinical significance in patients with esophagus and stomach double primary carcinoma(ESDC).Methods The expression of CDH1 gene methylation in cancerous tissues and adjacent cancerous tissues in 18 cases of ESDC were detected using methylation-specific PCR method. Results Eighteen patients were endoscopically diagnosed as ESDC between Jan. 2007 and Sep. 2009 in the 4th Hospital Affiliated to Hebei Medical University. The positive methylation of CDH1 gene in tissues of esophageal squamous cell carcinoma (ESCC)and adjacent cancer were 66.7% and 33. 3%, respectively, with significant difference (χ2= 4. 167, P = 0. 031). Whereas the positive methylation of CDH1 gene in tissues of gastric carcinoma (GA) and adjacent cancer were 77.8% and 44.4%, respectively, without statistical difference (χ2=1.786, P= 0. 180). There was no significant difference (P=0. 500) in positive rate of CDH1 gene methylation between ESCC tissues and GA tissues in same individual with ESDC. For 18 patients with ESDC, consistent change of CDH1 methylation in tissues of two kinds of cancers was found in 16 patients with a total agreement of 88.9 % (positive agreement of 66.7 % and negative agreement of 22. 2%). Statistical analysis showed a significant correlation between two groups (P = 0. 005). Conclusion In patients with ESDC, there is a high consistency of CDH1 methylation change, between ESCC and GA,which suggests that two kinds of cancer may have similar risk factors and molecular mechanisms.%目的 研究同一个体食管和胃双原发癌组织中CDH1基因启动子区CpG岛甲基化变化及其临床意义.方法 应用甲基化特异性PCR法,检测18例食管和胃双原发癌患者癌组织及癌旁组织中CDH1基因甲基化的表达.结果 2007年1月至2009年9月河北医科大学第四医院经内镜确诊18例双原发癌患者,食管鳞状细胞癌及癌旁组织CDH1基因

  19. Microarray-Based Analysis of Methylation Status of CpGs in Placental DNA and Maternal Blood DNA - Potential New Epigenetic Biomarkers for Cell Free Fetal DNA-Based Diagnosis

    DEFF Research Database (Denmark)

    Hatt, Lotte; Aagaard, Mads M; Graakjaer, Jesper;

    2015-01-01

    G sites to be used with methyl-sensitive restriction enzymes (MSRE) in PCR-based NIPD, we furthermore refined the list of selected sites, containing a restriction sites for one of 16 different methylation-sensitive restriction enzymes. We present a list of markers on chromosomes 13, 18 and 21...

  20. Aberrant DNA methylation in cervical carcinogenesis

    Institute of Scientific and Technical Information of China (English)

    Hui-Juan Yang

    2013-01-01

    Persistent infection with high-risk types of human papillomavirus(HPV) is known to cause cervical cancer; however,additional genetic and epigenetic alterations are required for progression from precancerous disease to invasive cancer.DNA methylation is an early and frequent molecular alteration in cervical carcinogenesis.In this review,we summarize DNA methylation within the HPV genome and human genome and identify its clinical implications.Methylation of the HPV long control region (LCR) and L1 gene is common during cervical carcinogenesis and increases with the severity of the cervical neoplasm.The L1 gene of HPV16 and HPV18 is consistently hypermethylated in invasive cervical cancers and can potentially be used as a clinical marker of cancer progression.Moreover,promoters of tumor suppressor genes (TSGs) involved in many cellular pathways are methylated in cervical precursors and invasive cancers.Some are associated with squamous cell carcinomas,and others are associated with adenocarcinomas.Identification of methylated TSGs in Pap smear could be an adjuvant test in cervical cancer screening for triage of women with high-risk HPV,atypical squamous cells of undetermined significance,or low grade squamous intraepithelial lesion (LSIL).However,consistent panels must be validated for this approach to be translated to the clinic.Furthermore,reversion of methylated TSGs using demethylating drugs may be an alternative anticancer treatment,but demethylating drugs without toxic carcinogenic and mutagenic properties must be identified and validated.

  1. Identifying differentially methylated genes using mixed effect and generalized least square models

    OpenAIRE

    2009-01-01

    Abstract Background DNA methylation plays an important role in the process of tumorigenesis. Identifying differentially methylated genes or CpG islands (CGIs) associated with genes between two tumor subtypes is thus an important biological question. The methylation status of all CGIs in the whole genome can be assayed with differential methylation hybridization (DMH) microarrays. However, patient samples or cell lines are heterogeneous, so their methylation pattern may be very different. In a...

  2. Methylation plotter: a web tool for dynamic visualization of DNA methylation data.

    Science.gov (United States)

    Mallona, Izaskun; Díez-Villanueva, Anna; Peinado, Miguel A

    2014-01-01

    Methylation plotter is a Web tool that allows the visualization of methylation data in a user-friendly manner and with publication-ready quality. The user is asked to introduce a file containing the methylation status of a genomic region. This file can contain up to 100 samples and 100 CpGs. Optionally, the user can assign a group for each sample (i.e. whether a sample is a tumoral or normal tissue). After the data upload, the tool produces different graphical representations of the results following the most commonly used styles to display this type of data. They include an interactive plot that summarizes the status of every CpG site and for every sample in lollipop or grid styles. Methylation values ranging from 0 (unmethylated) to 1 (fully methylated) are represented using a gray color gradient. A practical feature of the tool allows the user to choose from different types of arrangement of the samples in the display: for instance, sorting by overall methylation level, by group, by unsupervised clustering or just following the order in which data were entered. In addition to the detailed plot, Methylation plotter produces a methylation profile plot that summarizes the status of the scrutinized region, a boxplot that sums up the differences between groups (if any) and a dendrogram that classifies the data by unsupervised clustering. Coupled with this analysis, descriptive statistics and testing for differences at both CpG and group levels are provided. The implementation is based in R/shiny, providing a highly dynamic user interface that generates quality graphics without the need of writing R code. Methylation plotter is freely available at http://gattaca.imppc.org:3838/methylation_plotter/.

  3. A convenient method to generate methylated and un-methylated control DNA in methylation studies

    Directory of Open Access Journals (Sweden)

    Mehdi Manoochehri

    2013-09-01

    Full Text Available Methylated and un-methylated control DNA is an important part of DNA methylation studies. Although human and mouse DNA methylation control sets are commercially available, in case of methylation studies on other species such as animals, plants, and bacteria, control sets need to be prepared. In this paper a simple method of generating methylated and un-methylated control DNA is described. Whole genome amplification and enzymatic methylation were performed to generate un-methylated and methylated DNA. The generated DNA were confirmed using methylation sensitive/dependant enzymes, and methylation specific PCR. Control reaction assays confirmed the generated methylated and un-methylated DNA.

  4. 头颈胸部恶性肿瘤92例营养状况分析%Nutritional Status of 92 Patients With Head- neck and Thorax Carcinoma

    Institute of Scientific and Technical Information of China (English)

    于凤梅; 饶志勇; 柳园; 羊长青; 胡雯

    2011-01-01

    Objective To investigate the nutritional status of carcinoma patients with cross sectional survey and provide evidence for clinical nutrition therapy. Methods The nutritional status of 92 patients with head - neck and thorax carcinoma were evaluated by Subjective Global Assessment (SGA) , and were conducted laboratory examination, anthropometry and the measurements of body composition. Results The incidence rate of malnutri-tion was 29.4% by SGA, and it was differed in different cancer types and clinical stages, which were 22. 7% -37. 5% and 11.1% -38. 5% respectively. There was some correlation between SGA and most classic nutritional indexes (r =0. 212 -0. 422, P <0. 05) . Except total lymphocyte count and extracellular water, edema score in malnutrition patients was higher than those in patients with normal nutrition ( P < 0. 05 ) , and the rest indexes were lower ( P < 0. 05 ) . Conclusion Carcinoma patients have certain malnutrition, so early and continuous nutritional evaluation should be practised by many indexes in order to work out nutritionalscheme correspondingly.%目的 横断面调查头颈胸部肿瘤患者的营养状况,为临床营养治疗提供依据.方法 采用主观全面评价法(SGA)对92例头颈胸部恶性肿瘤患者进行营养调查,并测定生化指标、人体测量指标及人体成分指标.结果 92例头颈胸部恶性肿瘤患者SGA评定营养不良的发生率为29.4%,不同肿瘤类型及临床分期营养不良的发生率也不同,分别为22.7% ~ 37.5%和11.1% ~38.5%;SGA与多数传统营养指标之间有相关性(r=0.212 ~0.422,P<0.05);生化、人体测量及人体成分指标比较,除淋巴细胞总数及细胞外水无差异外,营养不良组水肿指数高于营养正常组(P<0.05),其他指标均低于营养正常组(P<0.05).结论 恶性肿瘤患者存在一定程度的营养不良,应结合多种指标早期持续地监测恶性肿瘤患者的营养状况,从而制定相应的营养治疗方案.

  5. 乳腺癌患者Runx3基因启动子甲基化状态及蛋白表达的关系%Relationship between methylation status of Runx3 gene promoter and Runx3 protein expression in patients with breast cancer

    Institute of Scientific and Technical Information of China (English)

    李冬霞; 武辉; 高建芝; 房志鑫

    2013-01-01

    目的:探讨乳腺癌组织中Runx3基因启动子甲基化及蛋白的表达与乳腺癌生物学行为的关系,并分析Runx3基因启动子区甲基化与其蛋白表达的相关性.方法:采用甲基化特异性PCR(methylation-specific PCR,MSP)技术和免疫组织化学SP法检测48例患者乳腺癌组织、18例癌旁乳腺组织中Runx3基因启动子甲基化情况及蛋白的表达.结果:Runx3基因启动子在乳腺癌组织及癌旁乳腺组织中的甲基化阳性率分别为52.1%和3.7% (P<0.05); Runx3蛋白在乳腺癌及癌旁乳腺组织中的阳性率分别为33.3%和92.6% (P<0.05);Runx3基因启动子甲基化及蛋白的表达分别与乳腺癌的组织学分级、临床分期及淋巴结转移有关(P<0.05);Runx3基因启动子甲基化与蛋白表达呈明显的负相关(P<0.05).结论:Runx3基因启动子的异常甲基化是引起蛋白表达缺失的主要原因,Runx3基因启动子的异常甲基化与乳腺癌的发生、发展有关.%Objective:To explore the relationship between methylation status of Runx3 gene promoter,Runx3 protein expression in patients with breast cancer and biological behavior of breast cancer,and analyze the correlation between methylation status of Runx3 gene promoter and Runx3 protein expression.Methods:Methylation-specific PCR (MSP) technique and immunohistchemical SP method were used to detect the methylation statuses of Runx3 gene promoter and Runx3 protein expressions in 48 cases of breast cancer tissue and 18 cases of breast tissue adjacent to breast cancer.Results:The positive methylation rates of Runx3 gene promoter in breast cancer tissue and breast tissue adjacent to breast cancer were 52.1% and 3.7%,respectively (P < 0.05) ; the positive rates of Runx3 protein in breast cancer tissue and breast tissue adjacent to breast cancer were 33.3% and 92.6%,respectively (P <0.05) ; the methylation status of Runx3 gene promoter and Runx3 protein expression were respectively

  6. Thrombocytosis portends adverse prognostic significance in patients with stage II colorectal carcinoma [v2; ref status: indexed, http://f1000r.es/4k6

    Directory of Open Access Journals (Sweden)

    Tianhua Guo

    2014-10-01

    Full Text Available Thrombocytosis portends adverse prognostic significance in many types of cancers including ovarian and lung carcinoma. In this study, we determined the prevalence and prognostic significance of thrombocytosis (defined as platelet count in excess of 400 × 103/μl in patients with colorectal cancer. We performed a retrospective analysis of 310 consecutive patients diagnosed at our Institution between 2004 and 2013. The patients (48.7% male and 51.3% female had a mean age of 69.9 years (+/- 12.7 years at diagnosis. Thrombocytosis was found in a total of 25 patients, with a higher incidence in those with stage III and IV disease (14.4% of patients. Although the mean platelet count increased with the depth of tumor invasion (pT, its values remained within normal limits in the whole patient cohort. No patient with stage I cancer (n=57 had elevated platelet count at diagnosis. By contrast, five of the 78 patients (6.4% with stage II cancer showed thrombocytosis, and four of these patients showed early recurrence and/or metastatic disease, resulting in shortened survival (they died within one year after surgery. The incidence of thrombocytosis increased to 12.2% and 20.6%, respectively, in patients with stage III and IV disease. The overall survival rate of patients with thrombocytosis was lower than those without thrombocytosis in the stage II and III disease groups, but this difference disappeared in patients with stage IV cancer who did poorly regardless of their platelet count. We concluded that thrombocytosis at diagnosis indicates adverse clinical outcome in colorectal cancer patients with stage II or III disease. This observation is especially intriguing in stage II patients because the clinical management of these patients is controversial. If our data are confirmed in larger studies, stage II colon cancer patients with thrombocytosis may be considered for adjuvant chemotherapy.

  7. Differing von Hippel Lindau genotype in paired primary and metastatic tumors in patients with clear cell renal cell carcinoma

    Directory of Open Access Journals (Sweden)

    Susan A.J. Vaziri

    2012-05-01

    Full Text Available In sporadic clear cell renal cell carcinoma (CCRCC, the von Hippel Lindau (VHL gene is inactivated by mutation or methylation in the majority of primary (P tumors. Due to differing effects of wild-type (WT and mutant (MT VHL gene on downstream signaling pathways regulating angiogenesis, VHL gene status could impact clinical outcome. In CCRCC, comparative genomic hybridization (CGH analysis studies have reported genetic differences between paired P and metastatic (M tumors. We thus sequenced the VHL gene in paired tumor specimens from 10 patients to determine a possible clonal relationship between the P tumor and M lesion(s in patients with CCRCC. Using paraffin embedded specimens, genomic DNA from microdissected samples (>80% tumor of paired P tumor and M lesions from all 10 patients, as well as in normal tissue from 6 of these cases, was analyzed. The DNA was used for PCR-based amplification of each of the 3 exons of the VHL gene. Sequences derived from amplified samples were compared to the wild-type VHL gene sequence (GeneBank Accession No. AF010238. Methylation status of the VHL gene was determined using VHL methylation-specific PCR primers after DNA bisulfite modification. In 4/10 (40% patients the VHL gene status differed between the P tumor and the M lesion. As expected, when the VHL gene was mutated in both the P tumor and M lesion, the mutation was identical. Further, while the VHL genotype differed between the primary tumor in different kidneys or multiple metastatic lesions in the same patient, the VHL germline genotype in the normal adjacent tissue was always wild-type irrespective of the VHL gene status in the P tumor. These results demonstrate for the first time that the VHL gene status can be different between paired primary and metastatic tissue in patients with CCRCC.

  8. 胃癌患者术前腹腔冲洗液中CDH1甲基化状态及其临床意义%Methylation status of CDH1 gene in preoperative abdominal lavage of patients with gastric cancer and its clinical significance

    Institute of Scientific and Technical Information of China (English)

    罗君; 王实; 凌志强; 王新保; 余齐鸣; 赵挺; 方仁桂; 王建军; 郑智国; 余江流; 方铣华

    2012-01-01

    目的 探讨术前腹腔冲洗液中CDH1基因的异常甲基化与胃癌进展的关系.方法 采用实时荧光甲基化特异性聚合酶链反应技术,检测92例胃癌患者术前腹腔冲洗液中CDH1基因启动子区域5′-CpG岛的甲基化状态,并分析其与临床病理因素及预后之间的关系.结果 92例胃癌患者中,有45例(48.9%)检测到了CDH1基因异常甲基化现象,其中完全甲基化12例(13.0%),部分甲基化33例(35.9%).CDH1基因甲基化与肿瘤大小、生长方式、分化程度、淋巴管侵犯、浸润深度、淋巴结转移、远处转移及临床分期有关(均P<0.05),而与性别、年龄、肿瘤部位及幽门螺杆菌感染无关(均P>0.05).CDH1甲基化与非甲基化患者术后中位无进展生存期分别为20和38个月,差异有统计学意义(P<0.01).经Cox模型分析证实,术前腹腔冲洗液DNA中的CDH1甲基化状态是影响胃癌患者术后生存的独立预后因素(P=0.000,RR=332.88, 95%CI:21.71~5105.07).结论 CDH1基因启动子区域5′-CpG岛的异常甲基化在胃癌中属于高频分子事件,术前腹腔冲洗液DNA中CDH1甲基化分析可反映胃癌进展状况.%Objective To explore the association between the progression of gastric cancer and the aberrant methylation of CDH1 gene in preoperative abdominal lavage fluid.Methods Real-time methylation-specific polymerase chain reaction(qMSP) was used to investigate the methylation status of the CDH1 gene promoter 5′-CpG islands from preoperative abdominal lavage fluid in 92 patients with gastric cancer.The associations between methylation of CDH1 genes and clinicopathologic features and prognosis were investigated.Results Among the 92 patients with gastric cancer,aberrant methylation of CDH1 gene was detected in 45 (48.9%) patients,including total aberrant methylation in 12 (13.0%) cases and partly aberrant methylation in 33 (35.9%) cases.Significant associatons were found between CDH1 methylation status

  9. DNA methylation and cognitive functioning in healthy older adults

    NARCIS (Netherlands)

    Schiepers, O.J.G.; Boxtel, van M.P.J.; Groot, R.H.M.; Jolles, J.; Kok, F.J.; Verhoef, P.; Durga, J.

    2012-01-01

    Long-term supplementation with folic acid may improve cognitive performance in older individuals. The relationship between folate status and cognitive performance might be mediated by changes in methylation capacity, as methylation reactions are important for normal functioning of the brain. Althoug

  10. Gene promoter methylation patterns throughout the process of cervical carcinogenesis

    NARCIS (Netherlands)

    Yang, Nan; Nijhuis, Esther R.; Volders, Haukeline H.; Eijsink, Jasper J. H.; Lendvai, Agnes; Zhang, Bo; Hollema, Harry; Schuuring, Ed; Wisman, G. Bea A.; van der Zee, Ate G. J.

    2010-01-01

    Objectives: To determine methylation status of nine genes, previously described to be frequently methylated in cervical cancer, in squamous intraepithelial lesions (SIL). Methods: QMSP was performed in normal cervix, low-grade ( L) SIL, high-grade (H) SIL, adenocarcinomas and squamous cell cervical

  11. Defining the cutoff value of MGMT gene promoter methylation and its predictive capacity in glioblastoma.

    Science.gov (United States)

    Brigliadori, Giovanni; Foca, Flavia; Dall'Agata, Monia; Rengucci, Claudia; Melegari, Elisabetta; Cerasoli, Serenella; Amadori, Dino; Calistri, Daniele; Faedi, Marina

    2016-06-01

    Despite advances in the treatment of glioblastoma (GBM), median survival is 12-15 months. O6-methylguanine-DNA methyltransferase (MGMT) gene promoter methylation status is acknowledged as a predictive marker for temozolomide (TMZ) treatment. When MGMT promoter values fall into a "methylated" range, a better response to chemotherapy is expected. However, a cutoff that discriminates between "methylated" and "unmethylated" status has yet to be defined. We aimed to identify the best cutoff value and to find out whether variability in methylation profiles influences the predictive capacity of MGMT promoter methylation. Data from 105 GBM patients treated between 2008 and 2013 were analyzed. MGMT promoter methylation status was determined by analyzing 10 CpG islands by pyrosequencing. Patients were treated with radiotherapy followed by TMZ. MGMT promoter methylation status was classified into unmethylated 0-9 %, methylated 10-29 % and methylated 30-100 %. Statistical analysis showed that an assumed methylation cutoff of 9 % led to an overestimation of responders. All patients in the 10-29 % methylation group relapsed before the 18-month evaluation. Patients with a methylation status ≥30 % showed a median overall survival of 25.2 months compared to 15.2 months in all other patients, confirming this value as the best methylation cutoff. Despite wide variability among individual profiles, single CpG island analysis did not reveal any correlation between single CpG island methylation values and relapse or death. Specific CpG island methylation status did not influence the predictive value of MGMT. The predictive role of MGMT promoter methylation was maintained only with a cutoff value ≥30 %.

  12. Epigenetic mechanisms in penile carcinoma

    DEFF Research Database (Denmark)

    Kuasne, Hellen; Marchi, Fabio Albuquerque; Rogatto, Silvia Regina

    2013-01-01

    Penile carcinoma (PeCa) represents an important public health problem in poor and developing countries. Despite its unpredictable behavior and aggressive treatment, there have only been a few reports regarding its molecular data, especially epigenetic mechanisms. The functional diversity in diffe......Penile carcinoma (PeCa) represents an important public health problem in poor and developing countries. Despite its unpredictable behavior and aggressive treatment, there have only been a few reports regarding its molecular data, especially epigenetic mechanisms. The functional diversity...... in different cell types is acquired by chromatin modifications, which are established by epigenetic regulatory mechanisms involving DNA methylation, histone acetylation, and miRNAs. Recent evidence indicates that the dysregulation in these processes can result in the development of several diseases, including...

  13. Microarray-Based Analysis of Methylation Status of CpGs in Placental DNA and Maternal Blood DNA--Potential New Epigenetic Biomarkers for Cell Free Fetal DNA-Based Diagnosis.

    Directory of Open Access Journals (Sweden)

    Lotte Hatt

    Full Text Available Epigenetic markers for cell free fetal DNA in the maternal blood circulation are highly interesting in the field of non-invasive prenatal testing since such markers will offer a possibility to quantify the amount of fetal DNA derived from different chromosomes in a maternal blood sample. The aim of the present study was to define new fetal specific epigenetic markers present in placental DNA that can be utilized in non-invasive prenatal diagnosis. We have conducted a high-resolution methylation specific beadchip microarray study assessing more than 450.000 CpG sites. We have analyzed the DNA methylation profiles of 10 maternal blood samples and compared them to 12 1st trimesters chorionic samples from normal placentas, identifying a number of CpG sites that are differentially methylated in maternal blood cells compared to chorionic tissue. To strengthen the utility of these differentially methylated CpG sites to be used with methyl-sensitive restriction enzymes (MSRE in PCR-based NIPD, we furthermore refined the list of selected sites, containing a restriction sites for one of 16 different methylation-sensitive restriction enzymes. We present a list of markers on chromosomes 13, 18 and 21 with a potential for aneuploidy testing as well as a list of markers for regions harboring sub-microscopic deletion- or duplication syndromes.

  14. TET family proteins and 5-hydroxymethylcytosine in esophageal squamous cell carcinoma.

    Science.gov (United States)

    Murata, Asuka; Baba, Yoshifumi; Ishimoto, Takatsugu; Miyake, Keisuke; Kosumi, Keisuke; Harada, Kazuto; Kurashige, Junji; Iwagami, Shiro; Sakamoto, Yasuo; Miyamoto, Yuji; Yoshida, Naoya; Yamamoto, Manabu; Oda, Shinya; Watanabe, Masayuki; Nakao, Mitsuyoshi; Baba, Hideo

    2015-09-15

    Mammalian DNA is epigenetically marked by 5'-cytosine methylation (5-methylcytosine [5-mC]). The Ten-eleven translocation (TET) enzymes (TET1, TET2, and TET3) are implicated in DNA demethylation, through dioxygenase activity that converts 5-mC to 5-hydroxymethylcytosine (5-hmC). Although decreased TET is reportedly associated with decreased 5-hmC levels in various cancers, functions of 5-hmC and TET expression in esophageal squamous cell carcinoma (ESCC) are unclear. We used ELISA and immunohistochemistry tests to analyze 5-hmC status in ESCC tissues, RT-qPCR to analyze TET family mRNA expression in normal and tumor tissues, and pyrosequencing to quantify LINE-1 (i.e., global DNA methylation) levels. ELISA and immunohistochemical testing showed 5-hmC levels were significantly lower in ESCC than in paired normal tissues (P ESCCs than paired normal tissues (P ESCCs (P = 0.003, r = 0.33). 5-hmC levels were also significantly associated with LINE-1 methylation level (P = 0.0002, r = 0.39). Patients with low 5-hmC levels had shorter overall survival than those with higher levels, although not significantly so (P = 0.084). In conclusion, 5-hmC expression was decreased in ESCC tissues, and was associated with TET2 expression level. TET2 reduction and subsequent 5-hmC loss might affect ESCC development.

  15. Parathyroid carcinoma

    DEFF Research Database (Denmark)

    Qvist, N; Krøll, L; Ladefoged, C;

    1986-01-01

    Parathyroid carcinoma is a slow growing tumor, and the patients most often die from complications to the hypercalcemia. Therefore, any attempt should be made to remove local recurrence and metastasis surgically, as medical treatment is disappointing. A case treated with extensive vascular surgery...

  16. Carcinoma vulvar

    Directory of Open Access Journals (Sweden)

    Yamit Peñas Zayas

    2015-11-01

    Full Text Available El carcinoma de la vulva tiene una incidencia de aproximadamente un 3-5% dentro de todas las enfermedades ginecológicas malignas. El 90% de los tumores malignos de la vulva está constituido por carcinoma epidermoide, el resto son adenocarcinomas, carcinomas de células basales y melanomas. Se realiza la presentación de un caso de una paciente femenina de 25 años de edad con antecedentes  de Diabetes Mellitus tipo II y trombopatia, que ingresa en el servicio de ginecología con un cuadro cutáneo polimorfo, localizado en labios mayores y menores, dado por lesiones eritematoerosivas y vegetante, sospechándose clínicamente el diagnóstico  de un carcinoma epidermoide, corroborándose el mismo histológicamente al realizarse biopsia de piel. Se indicó tratamiento con quimioterapia. Por la edad de la paciente y ser menos frecuente en mucosa que en la piel,  motivo la presentación del caso.

  17. 5-羟色胺转运体基因启动子区CpG岛甲基化状态与精神分裂症Ⅰ型和Ⅱ型的关联%Relationship Between Methylation Status of CpG Islands within the Promoter Region of Serotonin Transporter Gene and Type Ⅰ or Type Ⅱ Schizophrenia

    Institute of Scientific and Technical Information of China (English)

    张海生; 王伟; 余道军; 张顺泉; 谢健

    2011-01-01

    目的:探讨5-羟色胺转运体(5-HTT)基因启动子区CpG岛甲基化状态与精神分裂症Ⅰ型和Ⅱ型的关联.方法:运用特异性甲基检测PCR和直接测序法对62例精神分裂症Ⅰ型患者、38例Ⅱ型患者和50例健康被试5-HTT 基因启动子区CpG岛甲基化状态进行检测.结果:三组5-HTT基因启动子区CpC岛甲基化阳性率无显著性差异;精神分裂症Ⅰ型患者5-HTT基因启动子区CpC岛内位点甲基化率显著高于精神分裂症Ⅱ型患者和健康被试组.结论:5-HTT基因启动子区CpG岛内位点高甲基化可能是精神分裂症Ⅰ型的发病机制之一.%Objective: To explore the relationship between methylation status of CpG islands in the promoter region of serotonin transporter gene (5-HTT) and type Ⅰ or Ⅱ schizophrenia. Methods: Methylation Specific PCR and DNA sequencing were used to detect the methylation status of CpG islands in 62 type Ⅰ schizophrenia patients, 38 type Ⅱ schizophrenia patients and 50 health subjects. Results: The methylation rates of CpG islands within the promoter region of 5-HTT of the three groups are 52% (32/62), 47% (18/38) and 50% (25/50), respectively. The methylation rates of loci in CpG islands within the promoter region of 5-HTT in type Ⅰ schizophrenia group is significantly higher than those in type Ⅱ schizophrenia group and health subjects. Conclusion: The high methylation rate of loci in CpG islands within the promoter region of 5-HTT should be a possible factor to influence the nosogenesis of type Ⅰ schizophrenia.

  18. 鼻咽癌住院患者心理痛苦现状及影响因素分析%Evaluation of the status of psychosocial distress in nasopharyngeal carcinoma patients and analysis of its influencing factors

    Institute of Scientific and Technical Information of China (English)

    邵晓丽; 江锦芳

    2015-01-01

    Objective To investigate psychosocial distress status of hospitalized patients with nasopharyngeal carcinoma and then analyze its influencing factors.Methods Using the method of convenience sampling,the level of psychosocial distress of hospitalized patients with nasopharyngeal carcinoma was measured with psychosocial distress thermometer (DT),then by orderly classification Logistic regression analysis methods,the main factors affecting the degree of psychosocial distress were obtained.Results This study in cluded 235 patients,and 228 patients completed the survey effectively,DT score was (4.030±2.993) points,including DT < 4 points (mild pain) accounted for 36.8% (84/228),DT 4-6 points (moderate pain) accounted for 38.2% (87/228),DT≥7 points (severe pain) accounted for 25.0% (57/228).Regression analysis showed that the economic problems,emotional problems (depression,fear,sadness),body problems (eating,oral ulcer,sleep problems),age,education degree,stressful life events in the recent year,the time of disease diagnosis,and disease recurrence were the main factors influencing the degree of psychosocial distress.Conclusions Hospitalized patients with nasopharyngeal carcinoma have widespread psychosocial distress,clinical medical personnel should take individualized psychological evaluation and intervention according to the related factors of psychosocial distress in order to reduce the degree of psychosocial distress.%目的 调查鼻咽癌住院患者心理痛苦现状,分析其影响因素.方法 采用便利抽样的方法,使用心理痛苦温度计(Distress Thermometer,DT)测量鼻咽癌住院患者的心理痛苦水平,通过有序多分类Logistic回归分析方法,分析出影响其心理痛苦程度的主要因素.结果 本研究共纳入235例患者,有效完成问卷的患者为228例,心理痛苦得分为(4.030±2.993)分,其中DT<4分(轻度痛苦)者占36.8%(84/228),DT4~6分(中度痛苦)者占38.2%(87/228),DT≥7

  19. H3K4 dimethylation in hepatocellular carcinoma is rare compared with other hepatobiliary and gastrointestinal carcinomas and correlates with expression of the methylase Ash2 and the demethylase LSD1.

    Science.gov (United States)

    Magerl, Christian; Ellinger, Jörg; Braunschweig, Till; Kremmer, Elisabeth; Koch, Lin Kristin; Höller, Tobias; Büttner, Reinhard; Lüscher, Bernhard; Gütgemann, Ines

    2010-02-01

    Methylation of core histones regulates chromatin structure and gene expression. Recent studies have demonstrated that these methylation patterns have prognostic value for some tumors. Therefore, we investigated dimethylation of histone H3 at lysine 4 (H3K4diMe) and H3K4 methylating (Ash2 complex) and demethylating enzymes (LSD1) in carcinomas of the hepatic and gastrointestinal tract. High levels of H3K4diMe were rarely observed in 15.7% of hepatocellular carcinoma (8/51) unlike other carcinomas including, in ascending order, cholangiocellular carcinoma/adenocarcinoma of the extrahepatic biliary tract, gastric carcinoma, pancreatic ductal adenocarcinoma, and neuroendocrine carcinoma (P carcinomas (38/45) and correlated directly with H3K4diMe modification (correlation coefficient r = 0.53) and LSD1 expression (r = 0.35). In contrast to other carcinomas, 65.9% (29/44) of hepatocellular carcinomas analyzed showed no LSD1 expression (P carcinomas without LSD1 expression appeared to be frequently Ash2 and H3K4diMe weak or negative (P = .004). In summary, high H3K4diMe expression is rare in hepatocellular carcinoma compared with other carcinomas (negative predictive value 92.3%), which may aid in the differential diagnosis. Lack of H3K4diMe is possibly due to complex epigenetic regulation involving Ash2 and LSD1.

  20. Relationship between nucleosome positioning and DNA methylation

    Science.gov (United States)

    Chodavarapu, Ramakrishna K.; Feng, Suhua; Bernatavichute, Yana V.; Chen, Pao-Yang; Stroud, Hume; Yu, Yanchun; Hetzel, Jonathan; Kuo, Frank; Kim, Jin; Cokus, Shawn J.; Casero, David; Bernal, Maria; Huijser, Peter; Clark, Amander T.; Krämer, Ute; Merchant, Sabeeha S.; Zhang, Xiaoyu; Jacobsen, Steven E.; Pellegrini, Matteo

    2010-01-01

    Nucleosomes compact and regulate access to DNA in the nucleus, and are composed of approximately 147 bases of DNA wrapped around a histone octamer1, 2. Here we report a genome-wide nucleosome positioning analysis of Arabidopsis thaliana utilizing massively parallel sequencing of mononucleosomes. By combining this data with profiles of DNA methylation at single base resolution, we identified ten base periodicities in the DNA methylation status of nucleosome-bound DNA and found that nucleosomal DNA was more highly methylated than flanking DNA. These results suggest that nucleosome positioning strongly influences DNA methylation patterning throughout the genome and that DNA methyltransferases preferentially target nucleosome-bound DNA. We also observed similar trends in human nucleosomal DNA suggesting that the relationships between nucleosomes and DNA methyltransferases are conserved. Finally, as has been observed in animals, nucleosomes were highly enriched on exons, and preferentially positioned at intron-exon and exon-intron boundaries. RNA Pol II was also enriched on exons relative to introns, consistent with the hypothesis that nucleosome positioning regulates Pol II processivity. DNA methylation is enriched on exons, consistent with the targeting of DNA methylation to nucleosomes, and suggesting a role for DNA methylation in exon definition. PMID:20512117

  1. Dissociation dynamics of methylal

    Energy Technology Data Exchange (ETDEWEB)

    Beaud, P.; Frey, H.-M.; Gerber, T.; Mischler, B.; Radi, P.P.; Tzannis, A.-P. [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

    1999-08-01

    The dissociation of methylal is investigated using mass spectrometry, combined with a pyrolytic radical source and femtosecond pump probe experiments. Based on preliminary results two reaction paths of methylal dissociation are proposed and discussed. (author) 4 fig., 3 refs.

  2. Genetic and epigenetic alterations in hepatitis B virus-associated hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Yongjun; Tian; Jing-hsiung; James; Ou

    2015-01-01

    Hepatitis B virus(HBV) is a major cause of hepatocellular carcinoma(HCC). Its chronic infection can lead to chronic liver inflammation and the accumulation of genetic alterations to result in the oncogenic transformation of hepatocytes. HBV can also sensitize hepatocytes to oncogenic transformation by causing genetic and epigenetic changes of the host chromosomes. HBV DNA can insert into host chromosomes and recent large-scale whole-genome sequencing studies revealed recurrent HBV DNA integrations sites that may play important roles in the initiation of hepatocellular carcinogenesis. HBV can also cause epigenetic changes by altering the methylation status of cellular DNA, the post-translational modification of histones, and the expression of micro RNAs. These changes can also lead to the eventual hepatocellular transformation. These recent findings on the genetic and epigenetic alterations of the host chromosomes induced by HBV opened a new avenue for the development of novel diagnosis and treatments for HBV-induced HCC.

  3. Clinical observation about the impact of postoperative nutritional status on chemotherapy for ad- vanced gastric carcinoma%进展期胃癌术后营养状态对化疗影响的临床观察

    Institute of Scientific and Technical Information of China (English)

    尹小平; 朱栋良; 黎慰浩; 王芳元

    2014-01-01

    目的:观察进展期胃癌术后不同营养状态患者化疗期间的临床状况差异性,探讨营养治疗在营养不良患者化疗期间的临床意义。方法运用营养风险筛查(NRS2002)评价标准,选取进展期胃癌术后NRS>3分的80例患者,随机分为实验组和对照组,每组各40例。实验组患者进行肠外营养支持,对照组除不予肠外营养支持外其余同实验组。比较两组患者的营养生化指标、化疗毒副反应。结果实验组和对照组在营养生化指标、化疗毒副反应评分差异上有统计学意义(P<0.05)。结论营养不良状态是影响胃癌术后化疗的不利因素,降低化疗耐受力、生活质量,影响胃癌术后化疗的临床转归,营养支持治疗使营养不良患者化疗期间临床受益。%Objective To observe the clinical differences of postoperative nutritional status inchemotherapy patients with advanced gastric carcinoma and discuss the significance of nutritional therapy.Method According to the Nutritional Risk Screening 2002(NRS 2002),80 patients(NRS score >3)wererandomly divided into the experimental group(n =40)and control group(n =40).Patients in the experimental group received additional parenteral nutrition.Patients in the control group received the same treatment except for the parenteral nutrition.Nutritional biochemistry indexes,toxicities and side reactions ofthe chemotherapy were compared between the groups.Results There were significant differences in nutritional biochemistry indexes,toxicities and side reactions of the chemotherapy between the groups(P <0.05).Conclusion Poor nutrition status is a negative factor of postoperative chemotherapy for patientswith gastric carcinoma.It will reduce chemotherapy tolerance and quality of life,which has negative effectson the clinical outcomes of postoperative chemotherapy.Nutrition support will benefit those patients whohad malnutrition during the time of

  4. No evidence of oncogenic KRAS mutations in squamous cell carcinomas of the anogenital tract and head and neck region independent of human papillomavirus and p16(INK4a) status.

    Science.gov (United States)

    Prigge, Elena-Sophie; Urban, Katharina; Stiegler, Sandrine; Müller, Meike; Kloor, Matthias; Mai, Sabine; Ottstadt, Martine; Lohr, Frank; Wenz, Frederik; Wagner, Steffen; Wittekindt, Claus; Klussmann, Jens Peter; Hampl, Monika; von Knebel Doeberitz, Magnus; Reuschenbach, Miriam

    2014-11-01

    Carcinogenesis of squamous cell carcinomas (SCCs) in the anogenital tract and head and neck region is heterogeneous. A substantial proportion of SCC in the vulva, anus, and head and neck follows a human papillomavirus (HPV)-induced carcinogenic pathway. However, the molecular pathways of carcinogenesis in the HPV-independent lesions are not completely understood. We hypothesized that oncogenic Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations might represent a carcinogenic mechanism in a proportion of those HPV-negative cancers. Considering the repeated observation of KRAS-associated p16(INK4a) overexpression in human tumors, it was assumed that KRAS mutations might be particularly present in the group of HPV-negative, p16(INK4a)-positive cancers. To test this hypothesis, we analyzed 66 anal, vulvar, and head and neck SCC with known immunohistochemical p16(INK4a) and HPV DNA status for KRAS mutations in exon 2 (codons 12, 13, and 15). We enriched the tumor collection with HPV DNA-negative, p16(INK4a)-positive cancers. A subset of 37 cancers was also analyzed for mutations in the B-Raf proto-oncogene, serine/threonine kinase (BRAF) gene. None of the 66 tumors harbored mutations in KRAS exon 2, thus excluding KRAS mutations as a common event in SCC of the anogenital and head and neck region and as a cause of p16(INK4a) expression in these tumors. In addition, no BRAF mutations were detected in the 37 analyzed tumors. Further studies are required to determine the molecular events underlying HPV-negative anal, vulvar, and head and neck carcinogenesis. Considering HPV-independent p16(INK4a) overexpression in some of these tumors, particular focus should be placed on alternative upstream activators and potential downstream disruption of the p16(INK4a) pathway.

  5. Photodynamic therapy as adjunctive therapy for morpheaform basal cell carcinoma.

    Science.gov (United States)

    Torres, T; Fernandes, I; Costa, V; Selores, M

    2011-01-01

    The authors decided to evaluate the possible use of methyl-aminolevulinate photodynamic therapy (MAL-PDT) as adjunctive therapy for morpheaform basal cell carcinoma prior to standard surgical excision in order to reduce tumor size and volume and to facilitate surgical treatment. It was observed that MAL-PDT may be an option as an adjunctive therapy prior to standard surgical excision of morpheaform basal cell carcinoma, leading to less invasive surgery.

  6. Photodynamic therapy as adjunctive therapy for morpheaform basal cell carcinoma

    OpenAIRE

    Torres, T.; I. Fernandes; Costa, V.; Selores, M

    2011-01-01

    The authors decided to evaluate the possible use of methyl-aminolevulinate photodynamic therapy (MAL-PDT) as adjunctive therapy for morpheaform basal cell carcinoma prior to standard surgical excision in order to reduce tumor size and volume and to facilitate surgical treatment. It was observed that MAL-PDT may be an option as an adjunctive therapy prior to standard surgical excision of morpheaform basal cell carcinoma, leading to less invasive surgery.

  7. Indices of methylation in sperm DNA from fertile men differ between distinct geographical regions

    NARCIS (Netherlands)

    Consales, C.; Leter, G.; Bonde, J. P E; Toft, G.; Eleuteri, P.; Moccia, T.; Budillon, A.; Jönsson, B. A G; Giwercman, A.; Pedersen, H. S.; Ludwicki, J. K.; Zviezdai, V.; Heederik, D.; Spanò, M.

    2014-01-01

    STUDY QUESTION Which are the main determinants, if any, of sperm DNA methylation levels? SUMMARY ANSWER Geographical region resulted associated with the sperm methylation status assessed on genome-wide repetitive sequences. WHAT IS KNOWN ALREADY DNA methylation level, assessed on repetitive sequence

  8. Mixed Mucinous and Infiltrating Carcinoma Occurring in Male Breast- Study of Clinico-Pathological Features: A Rare Case Report.

    Science.gov (United States)

    Gupta, Kavita; Sharma, Swati; Kudva, Ranjini; Kumar, Sandeep

    2015-06-01

    Mucinous carcinoma is a less common histologic variant of breast cancer. Cases of mucinous carcinomas in male breast are extremely rare. Here, we describe a case of mixed mucinous carcinoma i.e. mucinous carcinoma with infiltrating ductal carcinoma component and showing apocrine differentiation in a 73-year-old man. This uncommon tumour entity has dismal prognosis and treatment depends largely on the tumour type, size, lymph node involvement and hormonal status.

  9. Investigation of MGMT and DAPK1 methylation patterns in diffuse large B-cell lymphoma using allelic MSP-pyrosequencing

    DEFF Research Database (Denmark)

    Kristensen, Lasse Sommer; Treppendahl, Marianne Bach; Asmar, Fazila;

    2013-01-01

    The tumor suppressor genes MGMT and DAPK1 become methylated in several cancers including diffuse large B-cell lymphoma (DLBCL). However, allelic methylation patterns have not been investigated in DLBCL. We developed a fast and cost-efficient method for the analysis of allelic methylation based...... on pyrosequencing of methylation specific PCR (MSP) products including a SNP. Allelic methylation patterns were reliably analyzed in standards of known allelic methylation status even when diluted in unmethylated DNA to below 1% methylation. When studying 148 DLBCL patients MGMT and DAPK1 methylation was observed...... in 19% and 89%, respectively, and among methylated and heterozygous patients 29% and 55%, respectively, were biallelically methylated. An association between the T-allele of the rs16906252 SNP and MGMT methylation was observed (p-value=0.04), and DAPK1 methylation of the A-allele was associated...

  10. Classification of Epstein-Barr virus-positive gastric cancers by definition of DNA methylation epigenotypes.

    Science.gov (United States)

    Matsusaka, Keisuke; Kaneda, Atsushi; Nagae, Genta; Ushiku, Tetsuo; Kikuchi, Yasuko; Hino, Rumi; Uozaki, Hiroshi; Seto, Yasuyuki; Takada, Kenzo; Aburatani, Hiroyuki; Fukayama, Masashi

    2011-12-01

    Epstein-Barr virus (EBV) is associated with Burkitt lymphoma, nasopharyngeal carcinoma, opportunistic lymphomas in immunocompromised hosts, and a fraction of gastric cancers. Aberrant promoter methylation accompanies human gastric carcinogenesis, though the contribution of EBV to such somatic methylation changes has not been fully clarified. We analyzed promoter methylation in gastric cancer cases with Illumina's Infinium BeadArray and used hierarchical clustering analysis to classify gastric cancers into 3 subgroups: EBV(-)/low methylation, EBV(-)/high methylation, and EBV(+)/high methylation. The 3 epigenotypes were characterized by 3 groups of genes: genes methylated specifically in the EBV(+) tumors (e.g., CXXC4, TIMP2, and PLXND1), genes methylated both in EBV(+) and EBV(-)/high tumors (e.g., COL9A2, EYA1, and ZNF365), and genes methylated in all of the gastric cancers (e.g., AMPH, SORCS3, and AJAP1). Polycomb repressive complex (PRC) target genes in embryonic stem cells were significantly enriched among EBV(-)/high-methylation genes and commonly methylated gastric cancer genes (P = 2 × 10(-15) and 2 × 10(-34), respectively), but not among EBV(+) tumor-specific methylation genes (P = 0.2), suggesting a different cause for EBV(+)-associated de novo methylation. When recombinant EBV was introduced into the EBV(-)/low-methylation epigenotype gastric cancer cell, MKN7, 3 independently established subclones displayed increases in DNA methylation. The promoters targeted by methylation were mostly shared among the 3 subclones, and the new methylation changes caused gene repression. In summary, DNA methylation profiling classified gastric cancer into 3 epigenotypes, and EBV(+) gastric cancers showed distinct methylation patterns likely attributable to EBV infection.

  11. Carcinoma verrugoso

    Directory of Open Access Journals (Sweden)

    Esteban Quesada Jiménez

    2004-09-01

    Full Text Available Se presenta el caso de un paciente masculino de 76 años, vecino de Turrialba, agricultor, que consultó por una lesión de 3 años de evolución, localizada en la palma de la mano derecha a nivel palmar y compromiso de los dedos de la misma mano, caracterizada como una neoformación exofítica verrugosa de 5 por 11 cm. aproximadamente, con material caseoso entre sus crestas. La lesión ha estado creciendo de forma acelerada en los últimos 3 meses, causándole dolor y que le imposibilita ellaborar. Se le realizaron exámenes y se descartaron varias causas infecciosas, y concluyendo luego de varias biopsias con el diagnóstico de un carcinoma verrugoso. El paciente fue tratado mediante una amputación parcial de la mano. Este tumor es una variante del carcinoma epidermoide y presentamos su clasificación, patogénesis, histopatología, manifestaciones clínicas más frecuentes y diagnóstico diferencial.A 76 year old farmer from Turrialba (Cartago, presented with a 3 year old lesion of his right pal and proximal fingers. It was exophitic, wart like, and it measured 5x11 cm, draining caseous material from its crests. The lesion had grows quickly for the last 3 months and it became tender to the point of making impossible for him to work. A series of tests were done to rule out other possible infections causes, after several biopsies the diagnosis of verrocous carcinoma was made. The patient underwent a partial amputation of his hand. This tumor is considered a from of squamous cell carcinoma, we present here its classification, pathogenesis, histopathology, clinical manifestations and diferential diagnosis.

  12. Regulation and function of DNA methylation in plants and animals

    KAUST Repository

    He, Xinjian

    2011-02-15

    DNA methylation is an important epigenetic mark involved in diverse biological processes. In plants, DNA methylation can be established through the RNA-directed DNA methylation pathway, an RNA interference pathway for transcriptional gene silencing (TGS), which requires 24-nt small interfering RNAs. In mammals, de novo DNA methylation occurs primarily at two developmental stages: during early embryogenesis and during gametogenesis. While it is not clear whether establishment of DNA methylation patterns in mammals involves RNA interference in general, de novo DNA methylation and suppression of transposons in germ cells require 24-32-nt piwi-interacting small RNAs. DNA methylation status is dynamically regulated by DNA methylation and demethylation reactions. In plants, active DNA demethylation relies on the repressor of silencing 1 family of bifunctional DNA glycosylases, which remove the 5-methylcytosine base and then cleave the DNA backbone at the abasic site, initiating a base excision repair (BER) pathway. In animals, multiple mechanisms of active DNA demethylation have been proposed, including a deaminase- and DNA glycosylase-initiated BER pathway. New information concerning the effects of various histone modifications on the establishment and maintenance of DNA methylation has broadened our understanding of the regulation of DNA methylation. The function of DNA methylation in plants and animals is also discussed in this review. © 2011 IBCB, SIBS, CAS All rights reserved.

  13. Temozolomide in aggressive pituitary adenomas and carcinomas

    OpenAIRE

    Leon D. Ortiz; Luis V. Syro; Scheithauer, Bernd W.; Fabio Rotondo; Humberto Uribe; Camilo E Fadul; Eva Horvath; Kalman Kovacs

    2012-01-01

    Temozolomide is an alkylating agent used in the treatment of gliomas and, more recently, aggressive pituitary adenomas and carcinomas. Temozolomide methylates DNA and, thereby, has antitumor effects. O 6-methylguanine-DNA methyltransferase, a DNA repair enzyme, removes the alkylating adducts that are induced by temozolomide, thereby counteracting its effects. A Medline search for all of the available publications regarding the use of temozolomide for the treatment of pituitary tumors was perf...

  14. 妇科恶性肿瘤患者主要照顾者心理健康状况的动态研究%A dynamic study on mental health status among primary caregivers of gynecological carcinoma patients

    Institute of Scientific and Technical Information of China (English)

    甘善英; 孔惠玲

    2015-01-01

    目的:调查妇科肿瘤患者主要照顾者心理健康状况,分析主要照顾者心理状况的动态变化与影响因素。方法对101例妇科肿瘤患者主要照顾者于患者手术前一天(T1)、手术后第3天(T2)、第1次化疗后(T3)、第3次化疗后(T4)进行动态调查分析。结果101名照顾者在患者病情不同阶段症状自评量表( SCL-90)总分和各因子分比较差异有统计学意义( P<0.05);家庭经济和患者疾病程度与SCL-90总分呈正相关,T2时点照顾者文化程度与SCL-90总分呈正相关;各个时点患者疾病程度均进入回归方程,显示与照顾者的心理健康状况独立相关。照顾者文化程度在T2时点进入了回归方程,家庭经济在T4时点进入了回归方程。结论医护人员要重视妇科恶性肿瘤患者主要照顾者的身心健康状况,从影响心理健康水平的各个方面制订针对性措施,促进其身心健康。%Objective To investigate the dynamic changes of mental health status among primary caregivers of gynecological carcino-ma patients. Methods The mental health status of the main caregivers of 101 patients with gynecologic tumor was investigated for 4 times (before surgery (T1), after surgery (T2), the first chemotherapy (T3), and the third chemotherapy (T4). Results In different stages the SCL-90 total score and each factor comparison of 101 caregivers in patients had statistical significance (P<0. 05);the correlation of SCL-90 total score in the four time points with family economy and degree of disease for patients had statistical significance. In T2 point the correla-tion of SCL-90 total score with caregivers' culture degree was statistically significant. Each point disease degree of patients entered the regres-sion equation, and the mental health status of the display and caregivers were associated independently. Caregivers′ cultural degree in T2 point entered the regression equation, ans family economy in T4 point entered the

  15. Molecular pathology of breast apocrine carcinomas

    DEFF Research Database (Denmark)

    Celis, J.E.; Gromova, I.; Gromov, P.;

    2006-01-01

    Breast cancer is a heterogeneous disease that encompasses a wide range of histopathological types including: invasive ductal carcinoma, lobular carcinoma, medullary carcinoma, mucinous carcinoma, tubular carcinoma, and apocrine carcinoma among others. Pure apocrine carcinomas represent about 0.5%...

  16. 非小细胞肺癌290例表皮生长因子受体基因突变和拷贝数的检测分析%Status of gene mutation and copy number of EGFR in 290 cases of non-small cell lung carcinoma

    Institute of Scientific and Technical Information of China (English)

    梁智勇; 曾瑄; 张静; 武莎斐; 高洁; 刘彤华

    2008-01-01

    目的 检测非小细胞肺癌(NSCLC)组织中表皮生长因子受体(EGFR)基因突变和拷贝数,明确中国NSCLC人群中EGFR基因状态.方法 应用显微切割技术和蝎形探针扩增阻滞突变系统(Scorpions ARMS)联合检测290例石蜡包埋、甲醛固定的NSCLC组织中其EGFR基因突变状态,并应用荧光原位杂交(FISH)技术检测其中209例EGFR基因拷贝数的情况,分析EGFR基因突变和EGFR摹因拷贝数二者之间的关系,探讨EGFR基因状态与肺癌临床病理特点的相关性.结果 NSCLC组织EGFR突变率为41.7%(121/290),其中腺癌为48.4%,大细胞癌为16.7%,鳞癌为0.EGFR基因突变以第19、21外显子突变为主(92/121),其中6例存在两种类型的突变.EGFR基因FISH阳性率为51.2%(107/209),包括29例扩增,78例高多体性.其中腺癌FISH阳性率为52.1%,大细胞癌为75.0%,鳞癌为11.1%.检测结果显示NSCLC组织EGFR基因突变与EGFR基因高拷贝数主要存在于腺癌,EGFR基因突变与EGFR基因拷贝数之间有显著的相关性.结论 中国人NSCLC组织具有较高的EGFR基因突变率和FISH阳性率;联合检测EGFR基因拷贝数和基因突变可能更有利于靶向药物的筛选.%Objective To investigate EGFR mutations and gene copy number status in non-small cell lung carcinomas in the Chinese patients. Methods Using formalin fixed and paraffin embedded tissue samples, EGFR mutations were investigated in 290 cases of non-small cell lung carcinomas by microdissection and scorpions amplification refractory mutation system. The status of EGFR gene copy number was investigated by FISH. Furthermore, the relationship between EGFR mutations and gene copy number, and the relationship between EGFR gene status and elinieopathologieal variables of non-small cell lung carcinoma were analyzed. Results The overall mutation rate of EGFR was 41.7% (121/290). The mutation rates in adenocarcinoma, large cell carcinoma and squamons carcinoma were 48.4%, 16.7% and O

  17. The correlation between CDH1 gene methylation status and clinical pathological characteristics and prognosis of colorectal cancer%CDH1基因甲基化状态与结直肠癌临床病理特征及预后的相关性分析

    Institute of Scientific and Technical Information of China (English)

    郭珊岚; 王卫

    2016-01-01

    目的:检测原发性结直肠癌患者术中腹腔灌洗液悬浮细胞中的CDH1基因启动子所在5'-CpG岛的异常甲基化,同时对其异常甲基化与临床病情发展、病理变化及术后预后的相关关系进行探讨。方法选取该院肿瘤科进行结直肠癌切除术的原发性结直肠癌患者,所有入选患者由专人进行跟踪随访。检测CDH1基因启动子所在5'-CpG岛的异常甲基化,对其异常甲基化与临床病情发展、病理变化及术后预后的相关关系进行探讨。结果该研究共纳入患者184例,其中甲基化组86例,非甲基化组98例。对两组患者临床病理结果检查可知甲基化组肿瘤直径大于非甲基化组(P<0.001),甲基化组肿瘤浸润性所占比例高于非甲基化组(P<0.001),甲基化组肿瘤分化程度低于非甲基化组(P<0.001),甲基化组淋巴转移率、远处转移率高于非甲基化组(P<0.001,P=0.026),甲基化组临床TNM分期更晚(P<0.001)。根据随访结果显示非甲基化组患者生存率高于甲基化组患者(P<0.05)。Cox比例风险模型结果显示,患者肿瘤的大体分型、分化程度、侵袭程度和病理分期是影响生存预后的变量,其中腹腔灌洗液悬浮细胞CDH1基因甲基化是影响预后最重要的独立因素,RR=28.514。结论原发性结直肠癌患者腹腔灌洗液悬浮细胞CDH1基因甲基化程度提高,恶性程度高,预后差。%Objective To detect the CDH1 gene methylation of suspension cells in intraoperative abdominal lavage fluid from colorectal cancer patients,and to examine its association with clinicopathology and prognosis.Methods Real-time methylation-specific poly-merase chain reaction(q-MSP)Was used to investigate the methylation status of the CDH1 gene promoter 5 ’-CpG islands from intra-operative abdominal lavage fluid in 184 patients with colorectal cancer.The associations

  18. Influence of DNA methyltransferase 3b on the expression of cylin D1 gene and methylation of its promoters in human hepatocellular carcinoma cells%DNA甲基转移酶3b对肝癌细胞系中细胞周期素D1基因的表达和启动子甲基化的影响

    Institute of Scientific and Technical Information of China (English)

    王佳辰; 王家祥; 刘怀然; 张勇敢

    2009-01-01

    Objeetive To investigate the influence of DNA methyltransferase(DNMT)3b on the expression of cylin D1 gene and methylation of its promoters and to investigate the function of DNMT3b.Methods Human hepatocellular carcinoma cells of the line SMMC7721 were cuhured and randomly divided into 3 groups:experimental group transfected with siRNA to silence the DNMT3b,control group transfected with control siRNA,and normal group without transfection.The transfection rate of siRNA was detected by fluorescence microscopy.MTr method was used to measure the survival rate of the SMMC-7721 cells.Western blotting and cell proliferation assay were performed to evaluate the expression of cyclin D1 and cell growth.Methylation specific PCR(MSP)was performed to investigate whether the promoter of cyclin D1 was methylated.Results F1uorescence microscopy showed that the transfection rate of siRNA was over 90%.MTr method showed that 24 h and 36 h after transfection the A value and survival rate of the SMMC7721cells of the experimental group were both significantly higher than those of the control d normal groups(all P<0.05).Western blotting showed that the expression levels of DNMl3b and cyclin D1 of the experimental groud decreased significantly compared with the control and the normal groups.MSP showed no obvious change of the state of methylation among the 3 groups.Condusions DNMT3b may regulate the expression and the function of cyclin D1 gene in the human hepatocellular carcinoma cells,but does not change its methylation state.DNMT3b may play their role as a signal transduction element rather than as a DNA methyltransferase.%目的 探讨DNA甲基转移酶3b(DNMT3b)在人肝癌细胞系(SMMC7721)中对细胞周期素D1(cylin D1)表达及其启动子甲基化水平的影响,并进一步探讨DNMT3b的作用.方法 用小分子干扰RNA(siRNA)技术抑制DNMT3b在SMMC7721细胞系中的表达(实验组),另设转染对照siRNA的对照组,及未加任何处理因素的正常组.用蛋

  19. Pathway Implications of Aberrant Global Methylation in Adrenocortical Cancer.

    Directory of Open Access Journals (Sweden)

    Christophe R Legendre

    Full Text Available Adrenocortical carcinomas (ACC are a rare tumor type with a poor five-year survival rate and limited treatment options.Understanding of the molecular pathogenesis of this disease has been aided by genomic analyses highlighting alterations in TP53, WNT, and IGF signaling pathways. Further elucidation is needed to reveal therapeutically actionable targets in ACC.In this study, global DNA methylation levels were assessed by the Infinium HumanMethylation450 BeadChip Array on 18 ACC tumors and 6 normal adrenal tissues. A new, non-linear correlation approach, the discretization method, assessed the relationship between DNA methylation/gene expression across ACC tumors.This correlation analysis revealed epigenetic regulation of genes known to modulate TP53, WNT, and IGF signaling, as well as silencing of the tumor suppressor MARCKS, previously unreported in ACC.DNA methylation may regulate genes known to play a role in ACC pathogenesis as well as known tumor suppressors.

  20. Human papillomavirus DNA and p16 expression in Japanese patients with oropharyngeal squamous cell carcinoma.

    Science.gov (United States)

    Kawakami, Hisato; Okamoto, Isamu; Terao, Kyoichi; Sakai, Kazuko; Suzuki, Minoru; Ueda, Shinya; Tanaka, Kaoru; Kuwata, Kiyoko; Morita, Yume; Ono, Koji; Nishio, Kazuto; Nishimura, Yasumasa; Doi, Katsumi; Nakagawa, Kazuhiko

    2013-12-01

    Human papillomavirus (HPV) is a major etiologic factor for oropharyngeal squamous cell carcinoma (OPSCC). However, little is known about HPV-related OPSCC in Japan. During the study, formalin-fixed, paraffin-embedded OPSCC specimens from Japanese patients were analyzed for HPV DNA by the polymerase chain reaction (PCR) and for the surrogate marker p16 by immuno-histochemistry. For HPV DNA-positive, p16-negative specimens, the methylation status of the p16 gene promoter was examined by methylation-specific PCR. Overall survival was calculated in relation to HPV DNA and p16 status and was subjected to multivariate analysis. OPSCC cell lines were examined for sensitivity to radiation or cisplatin in vitro. The study results showed that tumor specimens from 40 (38%) of the 104 study patients contained HPV DNA, with such positivity being associated with tumors of the tonsils, lymph node metastasis, and nonsmoking. Overall survival was better for OPSCC patients with HPV DNA than for those without it (hazard ratio, 0.214; 95% confidence interval, 0.074-0.614; P = 0.002). Multivariate analysis revealed HPV DNA to be an independent prognostic factor for overall survival (P = 0.015). Expression of p16 was associated with HPV DNA positivity. However, 20% of HPV DNA-positive tumors were negative for p16, with most of these tumors manifesting DNA methylation at the p16 gene promoter. Radiation or cisplatin sensitivity did not differ between OPSCC cell lines positive or negative for HPV DNA. Thus, positivity for HPV DNA identifies a distinct clinical subset of OPSCC with a more favorable outcome in Japanese.

  1. Analysis of nutritional status pre-and post-chemotherapy in patients with digestive carcinoma%消化系统肿瘤患者化疗前后营养状况变化分析

    Institute of Scientific and Technical Information of China (English)

    袁晓荣; 王亚兰; 孟令茹; 于焕欣; 杨雪洁; 许秀举

    2015-01-01

    Objective To explore the change in nutritional status pre-and post-chemotherapy on the digestive system carcinoma patients.Methods The data of 1 14 cases with digestive cancer were divided into two groups by gender,by age into three groups,young group,middle age group and elderly group,by disease stage into Ⅱ stage,Ⅲstage and Ⅳ stage group.Through anthropometric method and laboratory tests,the patients were monitored in triceps skinfold (TSF),ideal body mass (BW%),body mass index (BMI),hemoglobin (HGB),serum albumin (ALB), prealbumin (PA)and total lymphocyte count (TLC)pre-and post-chemotherapy with simple nutritional evaluation in different gender,different age groups and different disease stage groups.Results The gender showed no statistic significance in malnutrition rates (P > 0.05 ),with age increase,the malnutrition rates raised significantly after chemotherapy (P 0.05);随着年龄增长化疗后营养不良发生率明显增高(P <0.05);随分期的增高营养不良发生率明显增高(P <0.05)。结论①TSF 和 TLC 是相对较敏感指标,可以作为患者化疗前后营养不良评价指标;②化疗毒性对性别无选择性;③老年患者及病期较晚的患者对化疗的耐受性差,较中、青年患者及早期患者更易发生营养不良;④用 TSF 法评定为营养不良患者与 ALB、PA 及 TLC 评价结果的符合率较高,而与 HGB 评价结果符合率相对较低。

  2. O-6-methylguanine-deoxyribonucleic acid methyltransferase methylation enhances response to temozolomide treatment in esophageal cancer

    Directory of Open Access Journals (Sweden)

    Rifat Hasina

    2013-01-01

    Full Text Available Background: World-wide, esophageal cancer is a growing epidemic and patients frequently present with advanced disease that is surgically inoperable. Hence, chemotherapy is the predominate treatment. Cytotoxic platinum compounds are mostly used, but their efficacy is only moderate. Newer alkylating agents have shown promise in other tumor types, but little is known about their utility in esophageal cancer. Methods: We utilized archived human esophageal cancer samples and esophageal cancer cell lines to evaluate O-6-methylguanine-deoxyribonucleic acid methyltransferase (MGMT hypermethylation status and determined sensitivity to the alkylating drug temozolomide (TMZ. Immunoblot analysis was performed to determine MGMT protein expression in cell lines. To assess and confirm the effect of TMZ treatment in a methylated esophageal cancer cell line in vivo, a mouse flank xenograft tumor model was utilized. Results: Nearly 71% (12/17 of adenocarcinoma and 38% (3/8 of squamous cell carcinoma (SCC patient samples were MGMT hypermethylated. Out of four adenocarcinoma and nine SCC cell lines tested, one of each histology was hypermethylated. Immunoblot analyses confirmed that hypermethylated cell lines did not express the MGMT protein. In vitro cell viability assays showed the methylated Kyse-140 and FLO cells to be sensitive to TMZ at an IC 50 of 52-420 μM, whereas unmethylated cells Kyse-410 and SKGT-4 did not respond. In an in vivo xenograft tumor model with Kyse-140 cells, which are MGMT hypermethylated, TMZ treatment abrogated tumor growth by more than 60%. Conclusion: MGMT methylation may be an important biomarker in subsets of esophageal cancers and targeting by TMZ may be utilized to successfully treat these patients.

  3. [DNA methylation and epigenetics].

    Science.gov (United States)

    Vaniushin, B F

    2006-09-01

    In eukaryotic cells, nuclear DNA is subject to enzymatic methylation with the formation of 5-methylcytosine residues, mostly within the CG and CNG sequences. In plants and animals this DNA methylation is species-, tissue-, and organelle-specific. It changes (decreases) with age and is regulated by hormones. On the other hand, genome methylation can control hormonal signal. Replicative and post-replicative DNA methylation types are distinguished. They are mediated by multiple DNA methyltransferases with different site-specificity. Replication is accompanied by the appearance of hemimethylated DNA sites. Pronounced asymmetry of the DNA strand methylation disappears to the end of the cell cycle. A model of methylation-regulated DNA replication is proposed. DNA methylation controls all genetic processes in the cell (replication, transcription, DNA repair, recombination, and gene transposition). It is the mechanism of cell differentiation, gene discrimination and silencing. In animals, suppression of DNA methylation stops development (embryogenesis), switches on apoptosis, and is usually lethal. Disruption of DNA methylation pattern results in the malignant cell transformation and serves as one of the early diagnostic features of carcinogenesis. In malignant cell the pattern of DNA methylation, as well as the set of DNA methyltransferase activities, differs from that in normal cell. In plants inhibition of DNA methylation is accompanied by the induction of seed storage and florescence genes. In eukaryotes one and the same gene can be simultaneously methylated both at cytosine and adenine residues. It can be thus suggested, that the plant cell contains at least two different, and probably, interdependent systems of DNA methylation. The first eukaryotic adenine DNA methyltransferase was isolated from plants. This enzyme methylates DNA with the formation of N6-methyladenine residues in the sequence TGATCA (TGATCA-->TGm6ATCA). Plants possess AdoMet-dependent endonucleases

  4. Expression of ECRG4, a novel esophageal cancer-related gene,downregulated by CpG island hypermethylation in human esophageal squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    Chun-Mei Yue; Da-Jun Deng; Mei-Xia Bi; Li-Ping Guo; Shih-Hsin Lu

    2003-01-01

    AIM: To study the mechanisms responsible for inactivation of a novel esophageal cancer related gene 4 (ECRG4) in esophageal squamous cell carcinoma (ESCC). METHODS: A pair of primers was designed to amplify a 220 bp fragment, which contains 16 CpG sites in the core promoter region of the ECRG 4 gene. PCR products of bisulfite-modified CpG islands were analyzed by denaturing high-performance liquid chromatography (DHPLC), which were confirmed by DNA sequencing. The methylation status of ECRG 4 promoter in 20 cases of esophageal cancer and the adjacent normal tissues, 5 human tumor cell lines (esophageal cancer cell line-NEC, EC109, EC9706; gastric cancer cell line- GLC; human embryo kidney cell line-Hek293)and 2 normal esophagus tissues were detected. The expression level of the ECRG 4 gene in these samples was examined by RT-PCR. RESULTS: The expression level of ECRG 4 gene was varied.Of 20 esophageal cancer tissues, nine were unexpressed,six were lowly expressed and five were highly expressed compared with the adjacent tissues and the 2 normal esophageal epithelia. In addition, 4 out of the 5 human cell lines were also unexpressed. A high frequency of methylation was revealed in 12 (8 unexpressed and 4 lowly expressed)of the 15 (80%) downregulated cancer tissues and 3 of the 4 unexpressed cell lines. No methylation peak was observed in the two highly expressed normal esophageal epithelia and the methylation frequency was low (3/20) among the 20 cases in the highly expressed adjacent tissues. The methylation status of the samples was consistent with the result of DNA sequencing. CONCLUSION: These results indicate that the inactivation of ECRG 4gene by hypermethylation is a frequent molecular event in ESCC and may be involved in the carcinogenesis of this cancer.

  5. Epigenetic inactivation of secreted frizzled-related protein 2 in esophageal squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    Xiao-Wen Hao; Sheng-Tao Zhu; Yuan-Long He; Peng Li; Yong-Jun Wang; Shu-Tian Zhang

    2012-01-01

    AIM: To investigate the expression and methylation status of the secreted frizzled-related protein 2 (SFRP2) in esophageal squamous cell carcinoma (ESCC) and explore its role in ESCC carcinogenesis.METHODS: Seven ESCC cell lines (KYSE 30, KYSE150, KYSE410, KYSE510, EC109, EC9706 and TE-1) and one immortalized human esophageal epithelial cell line (Het-1A), 20 ESCC tissue samples and 20 paired adjacent non-tumor esophageal epithelial tissues were analyzed in this study. Reverse-transcription polymerase chain reaction (RT-PCR) was employed to investigate the expression of SFRP2 in cell lines, primary ESCC tumor tissue, and paired adjacent normal tissue. Methylation status was evaluated by methylation-specific PCR and bisulfite sequencing. The correlation between expression and promoter methylation of the SFRP2 gene was confirmed with treatment of 5-aza-2'-deoxycytidine. To assess the potential role of SFRP2 in ESCC, we established stable SFRP2-transfected cells and examined them with regard to cell proliferation, colony formation, apoptosis and cell cycle in vivo and in vitro.RESULTS: SFRP2 mRNA was expressed in the immortalized normal esophageal epithelial cell line but not in seven ESCC cell lines. By methylation-specific PCR, complete methylation was detected in three cell lines with silenced SFRP2 expression, and extensive methylation was observed in the other four ESCC cell lines. 5-aza-2'-deoxycytidine could restore the expression of SFRP2 mRNA in the three ESCC cell lines lacking SFRP2 expression. SFRP2 mRNA expression was obviously lower in primary ESCC tissue than in adjacent normal tissue (0.939 ± 0.398 vs 1.51 ± 0.399, P < 0.01). SFRP2 methylation was higher in tumor tissue than in paired normal tissue (95% vs 65%, P < 0.05). The DNA methylation status of the SFRP2 correlated inversely with the SFRP2 expression. To assess the potential role of SFRP2 in ESCC, we established stable SFRP2 transfectants and control counterparts by introducing pcDNA3.1/v5 his

  6. GATA3 expression in breast carcinoma: utility in triple-negative, sarcomatoid, and metastatic carcinomas.

    Science.gov (United States)

    Cimino-Mathews, Ashley; Subhawong, Andrea P; Illei, Peter B; Sharma, Rajni; Halushka, Marc K; Vang, Russell; Fetting, John H; Park, Ben Ho; Argani, Pedram

    2013-07-01

    GATA3 plays an integral role in breast luminal cell differentiation and is implicated in breast cancer progression. GATA3 immunohistochemistry is a useful marker of breast cancer; however, its use in specific subtypes is unclear. Here, we evaluate GATA3 expression in 86 invasive ductal carcinomas including triple-negative, Her-2, and luminal subtypes, in addition to 13 metaplastic carcinomas and in 34 fibroepithelial neoplasms. In addition, we report GATA3 expression in matched primary and metastatic breast carcinomas in 30 patients with known estrogen receptor (ER), progesterone receptor (PR), and Her-2 status, including 5 with ER and/or PR loss from primary to metastasis. Tissue microarrays containing 5 to 10 cores per tumor were stained for GATA3, scored as follows: 0 (0-5%), 1+ (6%-25%), 2+ (26%-50%), 3+ (51%-75%), and 4+ (>75%). GATA3 labeling was seen in 67% (66/99) of primary ductal carcinomas including 43% of triple-negative and 54% of metaplastic carcinomas. In contrast, stromal GATA3 labeling was seen in only 1 fibroepithelial neoplasm. GATA3 labeling was seen in 90% (27/30) of primary breast carcinomas in the paired cohort, including 67% of triple-negative carcinomas. GATA3 labeling was overwhelmingly maintained in paired metastases. Notably, GATA3 was maintained in all "luminal loss" metastases, which showed ER and/or PR loss. In conclusion, GATA3 expression is maintained between matched primary and metastatic carcinomas including ER-negative cases. GATA3 can be particularly useful as a marker for metastatic breast carcinoma, especially triple-negative and metaplastic carcinomas, which lack specific markers of mammary origin. Finally, GATA3 labeling may help distinguish metaplastic carcinoma from malignant phyllodes tumors.

  7. Outcome in unresectable glioblastoma: MGMT promoter methylation makes the difference.

    Science.gov (United States)

    Thon, Niklas; Thorsteinsdottir, Jun; Eigenbrod, Sabina; Schüller, Ulrich; Lutz, Jürgen; Kreth, Simone; Belka, Claus; Tonn, Jörg-Christian; Niyazi, Maximilian; Kreth, Friedrich Wilhelm

    2017-02-01

    In 2011, we reported a predominant prognostic/predictive role of MGMT promoter methylation status on progression-free survival (PFS) in unresectable glioblastoma patients undergoing upfront radiotherapy plus concomitant and maintenance temozolomide (RTX/TMZ → TMZ). We, here, present the final results of this prospective study focussing on the prognostic/predictive value of MGMT promoter methylation status for death risk stratification. Overall, 56 adult patients with unresectable, biopsy proven glioblastoma were prospectively assigned to upfront RTX/TMZ → TMZ treatment between March 2006 and August 2008. Last follow-up was performed in June 2016. MGMT promoter methylation was determined using methylation-specific PCR (MSP) and sodium bisulfite sequencing. Analyses were done by intention to treat. Prognostic factors were obtained from proportional hazard models. At the time of the final analysis 55 patients showed progressive disease and 53 patients had died. MGMT promoter was methylated (unmethylated) in 30 (26) patients. Methylation of the MGMT promoter was the strongest favorable predictor for overall survival (OS, median: 20.3 vs. 7.3 months, p MGMT promoter methylation status is essential for patients' counseling, prognostic evaluation, and for the design of future trials dealing with unresectable glioblastomas.

  8. Carcinoma verrugoso

    Directory of Open Access Journals (Sweden)

    Esteban Quesada Jiménez

    2004-09-01

    Full Text Available Se presenta el caso de un paciente masculino de 76 años, vecino de Turrialba, agricultor, que consultó por una lesión de 3 años de evolución, localizada en la palma de la mano derecha a nivel palmar y compromiso de los dedos de la misma mano, caracterizada como una neoformación exofítica verrugosa de 5 por 11 cm. aproximadamente, con material caseoso entre sus crestas. La lesión ha estado creciendo de forma acelerada en los últimos 3 meses, causándole dolor y que le imposibilita ellaborar. Se le realizaron exámenes y se descartaron varias causas infecciosas, y concluyendo luego de varias biopsias con el diagnóstico de un carcinoma verrugoso. El paciente fue tratado mediante una amputación parcial de la mano. Este tumor es una variante del carcinoma epidermoide y presentamos su clasificación, patogénesis, histopatología, manifestaciones clínicas más frecuentes y diagnóstico diferencial.

  9. Association of hypomethylation of LINE-1 repetitive element in blood leukocyte DNA with an increased risk of hepatocellular carcinoma

    Institute of Scientific and Technical Information of China (English)

    Jian-zhong DI; Xiao-dong HAN; Wen-ye GU; Yu WANG; Qi ZHENG; Pin ZHANG; Hui-min WU; Zhong-zheng ZHU

    2011-01-01

    Global DNA hypomethylation has been associated with increased risk for cancers of the colorectum,bladder,breast,head and neck,and testicular germ cells.The aim of this study was to examine whether global hypomethylation in blood leukocyte DNA is associated with the risk of hepatocellular carcinoma (HCC).A total of 315HCC cases and 356 age-,sex- and HBsAg status-matched controls were included.Global methylation in blood leukocyte DNA was estimated by analyzing long interspersed element-1 (LINE-1) repeats using bisulfite-polymerase chain reaction (PCR) and pyrosequencing.We observed that the median methylation level in HCC cases (percentage of 5-methylcytosine (5mC)=77.7%) was significantly lower than that in controls (79.5% 5mC) (P=0.004,Wilcoxon rank-sum test).The odds ratios (ORs) of HCC for individuals in the third,second,and first (lowest) quartiles of LINE-1methylation were 1.1 (95% confidence interval (CI) 0.7-1.8),1.4 (95% CI 0.8-2.2),and 2.6 (95% CI 1.7-4.1) (P for trend <0.001),respectively,compared to individuals in the fourth (highest) quartile.A 1.9-fold (95% CI 1.4-2.6) increased risk of HCC was observed among individuals with LINE-1 methylation below the median compared to individuals with higher (>median) LINE-1 methylation.Our results demonstrate for the first time that individuals with global hypomethylation measured in LINE-1 repeats in blood leukocyte DNA have an increased risk for HCC.Our data provide the evidence that global hypomethylation detected in the easily obtainable DNA source of blood leukocytes may help identify individuals at risk of HCC.

  10. Prognostic significance of stem cell marker CD133 determined by promoter methylation but not by immunohistochemical expression in malignant gliomas.

    Science.gov (United States)

    Wu, Xing; Wu, Fenlang; Xu, Dongwen; Zhang, Tao

    2016-04-01

    CD133 has played a pivotal role in the identification and isolation of brain tumor stem cells. The correlation between CD133 expression in tumor tissues with patients survival is still controversial. CD133 expression is determinated by methylation status of the promoter region 1-3. Aberrant methylation of CD133 was observed in glioblastoma. To date, a direct link between CD133 methylation and patient outcome has not been established.To address this question, we studied CD133 expression and promoter methylation in a series of 170 gliomas of various grade and histology, and investigated the correlation of CD133 expression and promoter methylation with patient outcome.We detected five CD133 promoter methylation patterns in 170 glioma samples: methylation only (M+, U-), unmethylation only (M-, U+), both methylation and unmethylation equally (M+, U+), high methylation and low unmethylation (M+, Ul), and low methylation and high unmethylation (Ml, U+). By multivariate survival analysis, we found CD133 promoter methylation status was significant (P promoter methylation status was observed (Kw = -0.165).CD133 promoter methylation status in glioma is closely correlated with patient survival, which suggest CD133 promoter methylaiton pattern is a promising tool for diagnostic purposes.

  11. 子宫颈鳞癌组织中表皮生长因子受体的表达及其基因状态分析%Analysis of epidermal growth factor receptor expression and gene expression status in tissue microarray of cervical squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    周素英; 冯国飞; 陈国荣; 潘丹; 张品南; 杨小敏; 夏作利

    2013-01-01

    目的 探讨宫颈鳞癌组织中表皮生长因子(EGFR)的表达及其基因状态,并分析EGFR的表达与基因状态之间的相关性.方法 构建含有72例宫颈鳞癌组织标本的组织芯片,应用免疫组化方法和荧光原位杂交(FISH)技术检测EGFR蛋白的表达、EGFR基因扩增、基因拷贝数的变化.结果 72例宫颈鳞癌中,因2例组织点发生脱片、移位,1例定位有误(未见癌细胞),剔除此3例,最终观察69例;EGFR蛋白表达的阳性率与肿瘤浸润深度、淋巴结转移及淋巴脉管间隙浸润有相关性(P均<0.05).FISH技术分析结果显示,72例宫颈鳞癌中有效信号共64例,基因扩增者7例,二倍体25例,三倍体23例,多倍体9例;所有EGFR基因扩增患者EGFR蛋白表达均为强阳性;EGFR蛋白表达与基因拷贝数增加之间有相关性(x2 =13.564,P<0.05).结论 EGFR蛋白过表达及基因状态可能参与宫颈鳞癌的生长、侵袭和转移过程,EGFR蛋白过表达可能由基因扩增或基因拷贝数增加所致.EGFR基因状态有可能成为宫颈鳞癌靶向治疗的有效监测指标.%Objective To elucidate the protein expression and gene expression status and the relationship between epidermal growth factor receptor (EGFR) protein expression and EGFR gene status.Methods Tissue microarray containing 72 cervical squamous cell carcinoma tissues was constructed,and EGFR protein expression and gene status were evaluated by immunohistochemical and fluorescence in situ hybridization (FISH) techniques.Results Protein expression of EGFR:69 of 72 cervical squamous cell carcinomas were observed.The results demonstrated it was significant association with invasion depth,lymphnode metastasis and lymph-vessel invasion (x2 =4.998,P < 0.05 ; x2 =4.299,P < 0.05 ; x2 =4.686,P < 0.05) in cervical squamous cell carcinomas.For FISH assessing EGFR gene,64 of 72 carcinomas were observed; 7 of 64 cases showed EGFR gene amplification,and 25 disomy,23 trisomy and 9 polysomy

  12. Effect of promoter methylation on lower expressed cyclooxygenase-2 esophageal squamous cell carcinoma xenograft%启动子甲基化调节在低表达环氧合酶-2的食管鳞癌移植模型中的作用

    Institute of Scientific and Technical Information of China (English)

    王青釭; 孟莹; 朱圣韬; 施海韵; 张澍田

    2014-01-01

    目的:环氧合酶-2(cyclooxygenase-2,COX-2)参与食管鳞癌(esophageal squamous cell carcinoma,ESCC)的发生发展过程,但在部分 ESCC 细胞株中 COX-2呈低表达。选择低表达 COX-2基因的 ESCC 进行裸鼠移植。探讨 COX-2基因甲基化与基因表达的关系,观察联合 COX-2选择性抑制剂和去甲基化药物干预对移植瘤生长的影响。方法选择 KYSE 150细胞进行裸鼠移植,采用数字表法随机分为4组:0.9%(质量分数)氯化钠注射液对照检查组7只。分别给予①尼美舒利(nimesulide,NIM)+5-氮杂脱氧胞苷(5-Aza-deoxycytidine,5-Aza)干预②尼美舒利干预③5-Aza 干预。5周后处死动物,比较各组移植瘤的直径,计算移植瘤体积,绘制生长曲线。反转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)测定组织中 COX-2 mRNA 表达;免疫组织化学方法测定 COX-2蛋白的表达;亚硫酸氢盐测序方法测定移植瘤中 COX-2启动子的甲基化状态。酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)法检测各组移植瘤中前列腺素 E2(prostaglandin E2,PGE2)的浓度。结果各组动物体质量增长差异无统计学意义,组间具有可比性。联合干预组移植瘤明显小于其他各组,差异具有统计学意义(P0. 05). Xenograft volumes were different between groups with that of the sodium chloride group smaller than other groups, and the difference was significant(P<0. 01). It was showed that methylation degree of COX-2 gene promoter was higher in NIM group and sodium choloride group than that in NIM combining 5-Aza group and 5-Aza group. For 10 CpG sites, methylation degrees were 30% and 58. 3% in group 3 and group 4, respectively. The expressions of COX-2 mRNA and protein were coincidently low in group 1 and high in group 3. PGE2 values were 0. 37, 0. 91 and 1. 22 in group 1, 2 and 3 compared with group 4, respectively. Conclusion Promoter methylation of COX-2 gene is one of the

  13. 先天性脊柱裂子鼠脊髓中微小RNA-124a基因甲基化状态研究%Methylation status of microRNA-124a in spinal cords of rats with congenital spina bifida

    Institute of Scientific and Technical Information of China (English)

    秦攀; 王家祥; 刘秋亮; 张大; 杨合英

    2014-01-01

    目的 检测微小RNA(miRNA)-124a在先天性脊柱裂子鼠脊髓中的表达及其启动子的甲基化水平.方法 应用实时荧光定量聚合酶链反应(FQ-PCR)方法检测42对子鼠脊髓标本中miRNA-124a的表达.随后,采用甲基化特异性PCR(MSP)检测样本中miRNA-124a上游启动子区的甲基化水平.结果 与正常对照组比较,脊柱裂子鼠脊髓中miRNA-124a的表达水平显著下调.miRNA-124a基因启动子高甲基化阳性率在脊柱裂子鼠脊髓及对照组中分别为81%和14%,差异有统计学意义(P<0.05).结论 先天性脊柱裂子鼠脊髓中miRNA-124a呈普遍的低表达趋势,其启动子区的高甲基化可能是导致miRNA-124a下调的主要因素,且可能与先天性脊柱裂发生有关.%Objective To investigate the expression of microRNA (miRNA)-124a and its function in spinal cord of rats with congenital spina bifida.Methods Forty-two couples of spinal cords were detec-ted by real-time fluorescent quantitative polymerase chain reaction (FQ-PCR).The DNA methylation sta-tus of miRNA-124a gene promoters was detected by DNA methylation specific polymerase chain reaction (MSP).Results As compared with normal controls,miRNA-124a was significantly decreased in rats with congenital spina bifida.The positive rate of DNA methylation of miRNA-124a gene promoter was 81% and 14% in CSB group and the normal control group,respectively.(P < 0.05).Conclusion miRNA-124a is down-regulated in majority of rats with congenital spina bifida,possibly because of the high methylation lev-el of its promoter.The high methylation of miRNA-124a might be involved in the occurrence of congenital spina bifida.

  14. A differentially methylated region of the DAZ1 gene in spermatic and somatic cells

    Institute of Scientific and Technical Information of China (English)

    Zuo-Xiang Li; Xu Ma; Zhao-Hui Wang

    2006-01-01

    Aim: To investigate the methylation status of the deleted in azoospermia 1 (DAZ1) gene promoter region in different cell types. Methods: Using CpG island Searcher software, a CpG island was found in the promoter region of the DAZ1 gene. The methylation status of this region was analyzed in sperm and leukocytes by bisulfited sequencing.Results: The methylation status of the CpG island in the DAZ1 gene promoter region differed in leukocytes and sperm: it was methylated in leukocytes, but unmethylated in sperm. Conclusion: A differentially methylated region of the DAZ1 gene exists in spermatic and somatic cells, suggesting that methylation of this region may regulate DAZ1 gene expression in different tissues.

  15. Identification and characterization of locus specific methylation patterns within novel loci undergoing hypermethylation during breast cancer pathogenesis

    DEFF Research Database (Denmark)

    Wojdacz, Tomasz K; Windeløv, Johanne Agerlin; Thestrup, Britta Boserup;

    2014-01-01

    biomarkers for early cancer detection and stratification of patients. METHODS: We used ultrahigh-resolution microarrays to compare genomewide methylation patterns of breast carcinomas (n = 20) and nonmalignant breast tissue (n = 5). Biomarker properties of a subset of discovered differentially methylated....... Analysis of the screening results showed that all DMRs tested displayed significant gains of methylation in the cancer tissue compared to the levels in control tissue. Interestingly, we observed two types of locus-specific methylation, with loci undergoing either predominantly full or heterogeneous...... methylation during carcinogenesis. Almost all tested DMRs (17 of 19) displayed low-level methylation in nonmalignant breast tissue, independently of locus-specific methylation patterns in cases. CONCLUSIONS: Specific loci can undergo either heterogeneous or full methylation during carcinogenesis, and loci...

  16. Influence of DNA methylation on positioning and DNA flexibility of nucleosomes with pericentric satellite DNA.

    Science.gov (United States)

    Osakabe, Akihisa; Adachi, Fumiya; Arimura, Yasuhiro; Maehara, Kazumitsu; Ohkawa, Yasuyuki; Kurumizaka, Hitoshi

    2015-10-01

    DNA methylation occurs on CpG sites and is important to form pericentric heterochromatin domains. The satellite 2 sequence, containing seven CpG sites, is located in the pericentric region of human chromosome 1 and is highly methylated in normal cells. In contrast, the satellite 2 region is reportedly hypomethylated in cancer cells, suggesting that the methylation status may affect the chromatin structure around the pericentric regions in tumours. In this study, we mapped the nucleosome positioning on the satellite 2 sequence in vitro and found that DNA methylation modestly affects the distribution of the nucleosome positioning. The micrococcal nuclease assay revealed that the DNA end flexibility of the nucleosomes changes, depending on the DNA methylation status. However, the structures and thermal stabilities of the nucleosomes are unaffected by DNA methylation. These findings provide new information to understand how DNA methylation functions in regulating pericentric heterochromatin formation and maintenance in normal and malignant cells.

  17. Giant basal cell carcinoma Carcinoma basocelular gigante

    Directory of Open Access Journals (Sweden)

    Nilton Nasser

    2012-06-01

    Full Text Available The basal cell carcinoma is the most common skin cancer but the giant vegetating basal cell carcinoma reaches less than 0.5 % of all basal cell carcinoma types. The Giant BCC, defined as a lesion with more than 5 cm at its largest diameter, is a rare form of BCC and commonly occurs on the trunk. This patient, male, 42 years old presents a Giant Basal Cell Carcinoma which reaches 180 cm2 on the right shoulder and was negligent in looking for treatment. Surgical treatment was performed and no signs of dissemination or local recurrence have been detected after follow up of five years.O carcinoma basocelular é o tipo mais comum de câncer de pele, mas o carcinoma basocelular gigante vegetante não atinge 0,5% de todos os tipos de carcinomas basocelulares. O Carcinoma Basocelular Gigante, definido como lesão maior que 5 cm no maior diâmetro, é uma forma rara de carcinoma basocelular e comumente ocorre no tronco. Este paciente apresenta um Carcinoma Basocelular Gigante com 180cm² no ombro direito e foi negligente em procurar tratamento. Foi realizado tratamento cirúrgico e nenhum sinal de disseminação ou recorrência local foi detectada após 5 anos.

  18. Reduced expression of Slit2 in renal cell carcinoma.

    Science.gov (United States)

    Ma, Wei-Jie; Zhou, Yu; Lu, Dan; Dong, Dong; Tian, Xiao-Jun; Wen, Jie-Xi; Zhang, Jun

    2014-01-01

    Slit2, initially identified as an important axon guidance molecule in the nervous system, was suggested to be involved in multiple cellular processes. Recently, Slit2 was reported to function as a potential tumor suppressor in diverse tumors. In this study, we systematically analyzed the expression level of Slit2 in renal cell carcinoma. Compared to paired adjacent non-malignant tissues, both Slit2 mRNA and protein expression were significantly down-regulated in renal cell carcinoma (RCC). Methylation-specific PCR showed that Slit2 promoter was methylated in two renal carcinoma cell lines. Pharmacologic demethylation dramatically induced Slit2 expression in cancer cell lines with weak expression of Slit2. Besides, bisulfite genomic sequencing confirmed that dense methylation existed in Slit2 promoter. Furthermore, in paired RCC samples, Slit2 methylation was observed in 8 out of 38 patients (21.1 %), which was well correlated with the down-regulation of Slit2 in RCC. Therefore, Slit2 may also be a potential tumor suppressor in RCC, which is down-regulated in RCC partially due to promoter methylation.

  19. Renal cell carcinoma with areas mimicking renal angiomyoadenomatous tumor/clear cell papillary renal cell carcinoma.

    Science.gov (United States)

    Petersson, Fredrik; Grossmann, Petr; Hora, Milan; Sperga, Maris; Montiel, Delia Perez; Martinek, Petr; Gutierrez, Maria Evelyn Cortes; Bulimbasic, Stela; Michal, Michal; Branzovsky, Jindrich; Hes, Ondrej

    2013-07-01

    We present a cohort of 8 renal carcinomas that displayed a variable (5%-95% extent) light microscopic appearance of renal angiomyoadenomatous tumor/clear cell papillary renal cell carcinoma (RAT/CCPRCC) without fulfilling the criteria for these tumors. All but 1 case predominantly (75%-95% extent) showed histopathologic features of conventional clear cell renal cell carcinoma. In 5 of 7 cases with mostly conventional clear renal cell carcinoma (CRCC) morphology, a diagnosis of CRCC was supported by the molecular genetic findings (presence of von Hippel-Lindau tumor suppressor [VHL] mutation and/or VHL promoter methylation and/or loss of heterozygosity [LOH] for 3p). Of the other 2 cases with predominantly characteristic CRCC morphology, 1 tumor did not reveal any VHL mutation, VHL promoter methylation, or LOH for 3p, and both chromosomes 7 and 17 were disomic, whereas the other tumor displayed polysomy for chromosomes 7 and 17 and no VHL mutation, VHL promoter methylation, or LOH for 3p. One tumor was composed primarily (95%) of distinctly RAT/CCPRCC-like morphology, and this tumor harbored a VHL mutation and displayed polysomy for chromosomes 7 and 17. Of the 5 cases with both histomorphologic features and molecular genetic findings of CRCC, we detected significant immunoreactivity for α-methylacyl-CoA racemase in 2 cases and strong diffuse immunopositivity for cytokeratin 7 in 3 cases. Despite the combination of positivity for α-methylacyl-CoA racemase and cytokeratin 7 in 2 cases, there was nothing to suggest of the possibility of a conventional papillary renal cell carcinoma with a predominance of clear cells.

  20. Methylated genes as new cancer biomarkers

    DEFF Research Database (Denmark)

    Brunner, Nils; Duffy, M.J; Napieralski, R.;

    2009-01-01

    Aberrant hypermethylation of promoter regions in specific genes is a key event in the formation and progression of cancer. In at least some situations, these aberrant alterations occur early in the formation of malignancy and appear to be tumour specific. Multiple reports have suggested that meas......Aberrant hypermethylation of promoter regions in specific genes is a key event in the formation and progression of cancer. In at least some situations, these aberrant alterations occur early in the formation of malignancy and appear to be tumour specific. Multiple reports have suggested...... that measurement of the methylation status of the promoter regions of specific genes can aid early detection of cancer, determine prognosis and predict therapy responses. Promising DNA methylation biomarkers include the use of methylated GSTP1 for aiding the early diagnosis of prostate cancer, methylated PITX2...... for predicting outcome in lymph node-negative breast cancer patients and methylated MGMT in predicting benefit from alkylating agents in patients with glioblastomas. However, prior to clinical utilisation, these findings require validation in prospective clinical studies. Furthermore, assays for measuring gene...

  1. Detection of MGMT promoter methylation in glioblastoma using pyrosequencing.

    Science.gov (United States)

    Xie, Hao; Tubbs, Raymond; Yang, Bin

    2015-01-01

    Recent clinical trials on patients with glioblastoma revealed that O6-Methylguanine-DNA methyltransferase (MGMT) methylation status significantly predicts patient's response to alkylating agents. In this study, we sought to develop and validate a quantitative MGMT methylation assay using pyrosequencing on glioblastoma. We quantified promoter methylation of MGMT using pyrosequencing on paraffin-embedded fine needle aspiration biopsy tissues from 43 glioblastoma. Using a 10% cutoff, MGMT methylation was identified in 37% cases of glioblastoma and 0% of the non-neoplastic epileptic tissue. Methylation of any individual CpG island in MGMT promoter ranged between 33% and 95%, with a mean of 65%. By a serial dilution of genomic DNA of a homogenously methylated cancer cell line with an unmethylated cell line, the analytical sensitivity is at 5% for pyrosequencing to detect MGMT methylation. The minimal amount of genomic DNA required is 100 ng (approximately 3,000 cells) in small fine needle biopsy specimens. Compared with methylation-specific PCR, pyrosequencing is comparably sensitive, relatively specific, and also provides quantitative information for each CpG methylation.

  2. p16INK4A and p14ARF Gene Promoter Hypermethylation as Prognostic Biomarker in Oral and Oropharyngeal Squamous Cell Carcinoma: A Review

    Science.gov (United States)

    Al-Kaabi, A.; van Bockel, L. W.; Pothen, A. J.; Willems, S. M.

    2014-01-01

    Head and neck squamous cell carcinoma is a heterogeneous group of tumors with each subtype having a distinct histopathological and molecular profile. Most tumors share, to some extent, the same multistep carcinogenic pathways, which include a wide variety of genetic and epigenetic changes. Epigenetic alterations represent all changes in gene expression patterns that do not alter the actual DNA sequence. Recently, it has become clear that silencing of cancer related genes is not exclusively a result of genetic changes such as mutations or deletions, but it can also be regulated on epigenetic level, mostly by means of gene promoter hypermethylation. Results from recent studies have demonstrated that DNA methylation patterns contain tumor-type-specific signatures, which could serve as biomarkers for clinical outcome in the near future. The topic of this review discusses gene promoter hypermethylation in oral and oropharyngeal squamous cell carcinoma (OSCC). The main objective is to analyse the available data on gene promoter hypermethylation of the cell cycle regulatory proteins p16INK4A and p14ARF and to investigate their clinical significance as novel biomarkers in OSCC. Hypermethylation of both genes seems to possess predictive properties for several clinicopathological outcomes. We conclude that the methylation status of p16INK4A is definitely a promising candidate biomarker for predicting clinical outcome of OSCC, especially for recurrence-free survival. PMID:24803719

  3. p16INK4A and p14ARF Gene Promoter Hypermethylation as Prognostic Biomarker in Oral and Oropharyngeal Squamous Cell Carcinoma: A Review

    Directory of Open Access Journals (Sweden)

    A. Al-Kaabi

    2014-01-01

    Full Text Available Head and neck squamous cell carcinoma is a heterogeneous group of tumors with each subtype having a distinct histopathological and molecular profile. Most tumors share, to some extent, the same multistep carcinogenic pathways, which include a wide variety of genetic and epigenetic changes. Epigenetic alterations represent all changes in gene expression patterns that do not alter the actual DNA sequence. Recently, it has become clear that silencing of cancer related genes is not exclusively a result of genetic changes such as mutations or deletions, but it can also be regulated on epigenetic level, mostly by means of gene promoter hypermethylation. Results from recent studies have demonstrated that DNA methylation patterns contain tumor-type-specific signatures, which could serve as biomarkers for clinical outcome in the near future. The topic of this review discusses gene promoter hypermethylation in oral and oropharyngeal squamous cell carcinoma (OSCC. The main objective is to analyse the available data on gene promoter hypermethylation of the cell cycle regulatory proteins p16INK4A and p14ARF and to investigate their clinical significance as novel biomarkers in OSCC. Hypermethylation of both genes seems to possess predictive properties for several clinicopathological outcomes. We conclude that the methylation status of p16INK4A is definitely a promising candidate biomarker for predicting clinical outcome of OSCC, especially for recurrence-free survival.

  4. Mixed and Ambiguous Endometrial Carcinomas: A Heterogenous Group of Tumors With Different Clinicopathologic and Molecular Genetic Features.

    Science.gov (United States)

    Espinosa, Iñigo; D'Angelo, Emanuela; Palacios, José; Prat, Jaime

    2016-07-01

    Besides endometrioid, serous, and clear cell carcinomas, there are endometrial carcinomas exhibiting mixed and ambiguous morphologic features. We have analyzed the immunophenotype (p53, p16, β-catenin, ER, HNF-1B, MLH1, and Ki-67) and mutational status (PTEN, KRAS, PIK3CA, and POLE) of 7 mixed carcinomas and 13 ambiguous carcinomas, all of them classified initially as mixed carcinomas. Only 2 of the 7 (28%) mixed carcinomas showed different immunophenotypes in different components. All but 2 tumors (5/7, 71%) overexpressed p53 and p16 and were negative for ER. Both carcinomas (2/7, 28%) showed a prominent micropapillary component that resembled an ovarian low-grade serous carcinoma and merged with villoglandular endometrioid carcinoma. The ambiguous carcinomas exhibited glandular architecture, high nuclear grade, and overlapping features of endometrioid and serous carcinomas. All tumors overexpressed p53 and p16, and the majority of cases (12/13, 92%) were negative for ER. KRAS mutations were identified in 3 of 7 (42%) mixed carcinomas, including the 2 cases with a "low-grade" serous-like component. PIK3CA mutations occurred in 2 (2/13, 15%) ambiguous carcinomas and PTEN mutations in 1 (1/7, 14%) mixed and 1 (1/13, 8%) ambiguous carcinoma. POLE exonuclease domain mutations were encountered in a case of mixed undifferentiated and well-differentiated (dedifferentiated) carcinoma. Two of the 7 (29%) mixed endometrial carcinomas and 5 of the 13 (38%) ambiguous carcinomas had extended beyond the pelvis (stages III and IV). Two of the 7 (29%) patients with mixed endometrial carcinoma and 6 of 12 (50%) patients with ambiguous endometrial carcinoma were alive with disease or had died of tumor. Our results show that, biologically, many so-called mixed carcinomas represent serous carcinomas with ambiguous morphology. Our series include 2 true mixed endometrial carcinomas with a "low-grade serous"-like component, microcystic, elongated, or fragmented features, KRAS mutations

  5. Current status and progress in studies on the association between obesity and DNA methylation among children and adolescents%儿童青少年肥胖与DNA甲基化关联研究进展

    Institute of Scientific and Technical Information of China (English)

    高莹; 高文静; 曹卫华

    2016-01-01

    DNA甲基化是目前研究最常见的表观遗传现象,有助于从生物学机制上解释遗传因素、环境因素及基因-环境因素交互作用对于肥胖的影响.生命早期DNA甲基化研究对于预防和控制肥胖具有重要作用,国内此方面研究尚缺乏.本研究通过回顾儿童青少年肥胖与DNA甲基化关联研究现状,介绍DNA甲基化研究的主要原理、基本特点及其与肥胖关联研究的主要结果,为今后相关研究提供依据.%DNA methylation is one of the most commonly recognized epigenetic phenomenon,which explains how genes,environmental factors and gene-environment interaction would cause obesity,integretedly.Studies on early life obesity-related epigenetic reveal important effects that related to the programs on prevention and control of obesity.However,only few similar studies have been conducted in China.This paper summarizes the basic principles,characteristics of DNA methylation and the major results of children and adolescents obesity-related research areas,in order to provide evidence for further studies.

  6. The in vitro biological activities of synthetic 18-O-methyl mycalamide B, 10-epi-18-O-methyl mycalamide B and pederin.

    Science.gov (United States)

    Richter, A; Kocienski, P; Raubo, P; Davies, D E

    1997-04-01

    Mycalamides A and B, which were originally isolated from a marine sponge, show close structural similarity to the insect toxin pederin, and exhibit potent cytotoxicity and antitumour activity. Detailed investigation of the clinical potential of these compounds has been hampered because they are available in only minute quantities from natural sources. We now describe the biological activities of 18-O-methyl mycalamide B, 10-epi-18-O-methyl mycalamide and pederin, all prepared by total synthesis. The activities of 18-O-methyl mycalamide B and pederin were virtually indistinguishable when evaluated in DNA or protein synthesis assays, and in cytotoxicity assays using human carcinoma cell lines (IC50s 0.2-0.6 nM). In all assays, 10-epi-18-O-methyl mycalamide B was 10(3) times less toxic than its diastereoisomer, demonstrating that the cytotoxicity of 18-O-methyl mycalamide B is inseparable from its ability to inhibit protein synthesis. Short-term exposure of squamous carcinoma cells to 18-O-methyl mycalamide B or pederin caused an irreversible inhibition of cellular proliferation and induced cellular necrosis. In contrast, the antiproliferative effects of the compounds on human fibroblasts were reversible and there was no evidence of necrosis. Demonstration that 18-O-methyl mycalamide B and the synthetically less complex molecule, pederin, show some tumour cell toxicity indicates that this novel class of compounds should be subjected to preclinical evaluation.

  7. P16 METHYLATION OF THE COLORECTAL CANCER AND ASSOCIATION WITH DUKES STAGES

    Institute of Scientific and Technical Information of China (English)

    王志伟; 易静; 仓辉; 邹鸿志; 郁宝铭; 汤雪明

    2001-01-01

    To explore whether methylation of the CpG island in the promoter of the p16 tumor suppressor gene was associated with clinicopathological characteristics of the colorectal cancer patients. Methods: Methylation- specific PCR (MSP) was used to detect p16 methylation of the colorectal cancer patients. Results: In 58 sporadic colorectal cancer, 43.1% of the tumors had detectable p16 methylation. Dukes' stage was associated with p16 methylation status. Dukes C, D patients (75%) were more likely to contain methylated p16 compared with Dukes A, B patients (13.3%). Conclusion: p16 methylation plays a role in the carcinogenesis of a subset of colorectal cancer. P16 methylation might be considered as a prognostic indicator.

  8. Thyroid cancer - medullary carcinoma

    Science.gov (United States)

    Thyroid - medullary carcinoma; Cancer - thyroid (medullary carcinoma); MTC; Thyroid nodule - medullary ... in children and adults. Unlike other types of thyroid cancer, MTC is less likely to be caused by ...

  9. Stages of Adrenocortical Carcinoma

    Science.gov (United States)

    ... Symptoms of adrenocortical carcinoma include pain in the abdomen. These and other signs and symptoms may be caused by adrenocortical carcinoma: A lump in the abdomen . Pain the abdomen or back. A feeling of ...

  10. Undifferentiated salivary gland carcinomas

    DEFF Research Database (Denmark)

    Herbst, H.; Hamilton-Dutoit, S.; Jakel, K.T.;

    2004-01-01

    Undifferentiated salivary gland carcinomas may be divided into small cell and large cell types. Among large cell undifferentiated carcinomas, lymphoepithelial carcinomas have to be distinguished, the latter of which are endemic in the Arctic regions and southern China where virtually all cases...... of these tumors are associated with the Epstein-Barr virus (EBV). Association with EBV may also be observed in sporadic cases, and detection of EBV gene products may aid their diagnosis. Immunohistology may be employed to resolve the differential diagnosis of undifferentiated salivary gland carcinomas, comprising...... malignant lymphomas, amelanotic melanomas, Merkel cell carcinomas, and adenoid cystic carcinomas, in particular in small biopsy materials. Because of the rarity of undifferentiated salivary gland carcinomas, the differential diagnosis should always include metastases of undifferentiated carcinomas arising...

  11. Liver cancer - hepatocellular carcinoma

    Science.gov (United States)

    Primary liver cell carcinoma; Tumor - liver; Cancer - liver; Hepatoma ... Hepatocellular carcinoma accounts for most liver cancers. This type of cancer occurs more often in men than women. It is usually diagnosed in people age 50 or ...

  12. The possible role of cell cycle regulators in multistep process of HPV-associated cervical carcinoma

    Directory of Open Access Journals (Sweden)

    Moneir Manar

    2007-05-01

    Full Text Available Abstract Background Human papillomavirus (HPV 16 and 18 are associated with cervical carcinogenesis through an interaction between HPV oncogenic proteins and cell cycle regulatory genes. However, the exact pathogenetic mechanisms are not determined yet. Methods We investigated 43 invasive squamous cell carcinoma (ISCC, 38 CIN III, 11 CINII and 18 CINI for cyclin D1, cyclin E, CDK4, p53, mdm-2, p21waf, p27, p16INK4A, Rb and Ki-67 aberrations using immunohistochemistry and molecular techniques. Twenty samples of normal cervical tissues (NCT were taken as a control. Results There was a significant increase in the expression of Ki-67, cyclin E, CDK4, p16INK4A, Rb (p= 0.003, 0.001, 0.001, 0.01 and a significant decrease in p27KIP1 from NCT to ISCC (p = 0.003. Increased cyclin D1, p21waf, p53, mdm-2 expression, homozygous deletion (HZD and promoter methylation (PM of the Rb were detected in CINIII and ISCC only. On univariate analysis; tumor size, differentiation, lymph node status, FIGO stage, Ki- 67, cyclin D1, p53 and p27KIP1 are significantly associated with reduced overall survival (OS while on multivariate analysis; only FIGO stage, Ki-67, cyclin D1, p53 and p27KIP1 were significant. Conclusion 1 Aberrations involving p27KIP1, cyclin E, CDK4, p16INK4A are considered early events in HPV 16 and 18-associated cervical carcinoma, whereas cyclin D1 and p53 pathway abnormalities are considered late events. 2 Immunohistochemical tests for p16INK4Aand cyclin E, could help in early diagnosis of cervical carcinoma. 3 Only FIGO stage p53, cyclin D1, p27KIP1 and Ki-67 are independent prognostic factors that might help in predicting outcome of cervical cancer patients.

  13. (123)I-interleukin-2 uptake in squamous cell carcinoma of the head and neck carcinoma

    NARCIS (Netherlands)

    Loose, David; Signore, Alberto; Staelens, Ludovicus; Bulcke, Katia Vanden; Vermeersch, Hubert; Dierckx, Rudi Andre; Bonanno, Elena; de Wiele, Christophe Van

    2008-01-01

    Introduction Information obtained on the IL-2 receptor status of tumour infiltrating lymphocytes in patients suffering from squamous cell carcinoma of the head and neck (SSCHN) before and after IL-2 treatment may lead to a better understanding of the immunological changes and related kinetics induce

  14. Clinicopathological Features of Triple Negative Breast Carcinoma

    Science.gov (United States)

    Reddy, Gowry Maram; Pai, Radha R.

    2017-01-01

    Introduction Breast carcinoma is one of the most common malignancies affecting women in developing countries. Molecular studies of breast carcinoma have classified the tumour based on the immunohistochemical staining into 4 subtypes, such as Luminal A, Luminal B, HER2/neu Positive and Triple Negative Breast Carcinoma (TNBC). TNBCs are reported to have an aggressive behaviour and wide metastasis, leading to selective treatment outcomes. Aim The aim was to study the clinicopathological features such as age, site, tumour size, histopathological type, histologic grade, lymph node status, stage and treatment outcomes of triple negative breast carcinoma. Materials and Methods A retrospective study was conducted on 108 cases of breast carcinoma received during the period of 2 years. The tumour was classified based on immunohistochemical staining into four subtypes. The clinicopathological details, histomorphological and immunohistochemical features of TNBC were studied. Results Of the 108 patients, 34 patients were diagnosed as TNBC. The average age at presentation was 48 years. Most of the cases showed Nottingham Modification of Scarff Bloom-Richardson (NMBR) grade 3 (55.9%) and stage II (67.6%). Ly-mph node metastasis was seen in 50% of cases. Infiltrating ductal carcinoma (not otherwise specified) type (91.2%) was the most common histological type. Among the other subtypes, Luminal A carcinoma was the most common (36.1%), followed by TNBC (31.5%) and HER2/neu positive carcinomas (28.7%). Compared to the other types of tumours, TNBC showed the most frequent distant lymph node metastasis (50%) when compared to luminal A (38.5%), luminal B (25%), HER2/neu positive (48.4%). Unlike the other types of tumours, TNBC were mostly high-grade. Conclusion TNBC have an aggressive behaviour compared to other subtypes with higher NMBR grade, nuclear pleomorphism, high mitotic rate and lymph node metastasis. PMID:28273970

  15. Quantitative evaluation of DNMT3B promoter methylation in breast cancer patients using differential high resolution melting analysis.

    Science.gov (United States)

    Naghitorabi, M; Mohammadi Asl, J; Mir Mohammad Sadeghi, H; Rabbani, M; Jafarian-Dehkordi, A; Javanmard, Haghjooye S

    2013-07-01

    DNA methylation plays an important role in carcinogenesis through epigenetic silencing of tumor suppressor genes. Aberrant methylation usually results from changes in the activity of DNA methyltransferases (DNMTs). Some studies show that the overexpression of the DNMTs may lead to aberrant methylation of tumor suppressor genes. Also the overexpression of DNMTs may be related to methylation status of their genes. Due to limited number of studies on DNMT3B promoter methylation, this study was performed to quantitatively measure the methylation level of DNMT3B gene in archival formalin fixed paraffin embedded (FFPE) tissues from breast cancer patients. Using differential high resolution melting analysis (D-HRMA) technology, the methylation level of DNMT3B gene promoter was quantified in 98 breast cancer FFPE tissues and also 10 fresh frozen normal tissue samples. Statistical analyses used for analyzing the correlation between the methylation and clinical variables. All the normal samples were found to be methylated at the DNMT3B promoter (the average methylation level 3.34%). Patients were identified as hypo-methylated (mean methylation level 0.8%), methylated (mean methylation level 2.48%) and hyper-methylated (mean methylation level 10.5%). Statistical analysis showed a significant correlation between the methylation status and the sample type, cancer type and tumor size. Also the methylation level was significantly associated with histologic grade. It is concluded that quantification of DNMT3B promoter methylation might be used as a reliable and sensitive diagnostic and prognostic tool in breast cancer. Also D-HRMA is demonstrated as a rapid and cost effective method for quantitative evaluation of promoter methylation.

  16. ENZYMOLOGY OF ARSENIC METHYLATION

    Science.gov (United States)

    Enzymology of Arsenic MethylationDavid J. Thomas, Pharmacokinetics Branch, Experimental Toxicology Division, National Health and Environmental Effects Research Laboratory, Office of Research and Development, U.S. Environmental Protection Agency, Research Triangle Park...

  17. Lysine methylation: beyond histones

    Institute of Scientific and Technical Information of China (English)

    Xi Zhang; Hong Wen; Xiaobing Shi

    2012-01-01

    Posttranslational modifications (PTMs) of histone proteins,such as acetylation,methylation,phosphorylation,and ubiquitylation,play essential roles in regulating chromatin dynamics.Combinations of different modifications on the histone proteins,termed 'histone code' in many cases,extend the information potential of the genetic code by regulating DNA at the epigenetic level.Many PTMs occur on non-histone proteins as well as histones,regulating protein-protein interactions,stability,localization,and/or enzymatic activities of proteins involved in diverse cellular processes.Although protein phosphorylation,ubiquitylation,and acetylation have been extensively studied,only a few proteins other than histones have been reported that can be modified by lysine methylation.This review summarizes the current progress on lysine methylation of nonhistone proteins,and we propose that lysine methylation,like phosphorylation and acetylation,is a common PTM that regulates proteins in diverse cellular processes.

  18. DNA methylation and not H3K4 trimethylation dictates the expression status of miR-152 gene which inhibits migration of breast cancer cells via DNMT1/CDH1 loop.

    Science.gov (United States)

    Sengupta, Dipta; Deb, Moonmoon; Rath, Sandip Kumar; Kar, Swayamsiddha; Parbin, Sabnam; Pradhan, Nibedita; Patra, Samir Kumar

    2016-08-15

    MicroRNAs (miRNA) are small non-coding RNAs which targets most protein-coding transcripts (mRNA) and destroy them. Thus miRNA controls the abundance of those specific proteins and impact on developmental, physiological and pathological processes. Dysregulation of miRNA function thus may lead to various clinicopathological complications, including breast cancer. Silencing of miR-152 gene due to promoter DNA methylation alter the expression pattern of several other genes. E-cadherin (CDH1) forms the core of adherent junctions between surrounding epithelial cells, link with actin cytoskeleton and affects cell signaling. CDH1 gene is down regulated by promoter DNA methylation during cancer progression. In this investigation, we attempt to elucidate the correlation of miR-152 and CDH1 function, as it is well known that the loss of CDH1 function is one of the major reasons for cancer metastasis and aggressiveness of spreading. For the first time we have shown that loss of CDH1 expression is directly proportional to the loss of miR-152 function in breast cancer cells. mRNA and protein expression profile of DNMT1 implicate that miR-152 targets DNMT1 mRNA and inhibits its protein expression. Tracing the molecular marks on DNA and histone 3 for understanding the mechanism of gene regulation by ChIP analyses leads to a paradoxical result that shows DNA methylation adjacent to active histone marking (enrichment of H3K4me3) silence miR-152 gene. Further experiments revealed that DNMT1 plays crucial role for regulation of miR-152 gene. When DNMT1 protein function is blocked miR-152 expression prevails and destroys the mRNA of DNMT1; this molecular regulatory mechanism is creating a cyclic feedback loop, which is now focused as DNMT1/miR-152 switch for on/off of DNMT1 target genes. We discovered modulation of CDH1 gene expression by DNMT1/miR-152 switches. We have demonstrated further that DNMT1 down regulation mediated upregulation of CDH1 (hereafter, DNMT1/CDH1 loop) in

  19. Deoxyribonucleic acid (DNA) methyltransferase contributes to p16 promoter CpG island methylation in lung adenocarcinoma with smoking.

    Science.gov (United States)

    Sun, Rongju; Liu, Jiahong; Wang, Bo; Ma, Lingyun; Quan, Xiaojiao; Chu, Zhixiang; Li, Tanshi

    2015-01-01

    In this study, the relationship between CpG island methylation and smoking and DNA methyltransferase in the occurrence and development of lung adenocarcinoma was explored by detecting p16 promoter methylation status. Protein and mRNA levels of p16 were detected by immunohistochemistry and in situ hybridization assays. p16 gene promoter and exon 1 CpG island locus Hap II sites methylation status was analyzed with the methylation-specific PCR. Only 4 of 40 p16-positive cases were detected to methylate on CpG islands with 10% methylating rate whereas 18 of p16-negative cases were methylated up to 36.73% of methylating rate. The methylating rates of both p16-positive and p16-negative groups were significantly different. 17 of 50 cases with smoking from total 89 lung adenocarcinoma cases were detected to methylate on CpG islands while only 5 of the remaining 39 non-smokers to methylate. The difference of the methylating rates in both smokers and non-smokers was significant to suggest the closely association of CpG island methylation of p16 with smoking. Furthermore, p16 promoter CpG islands were detected to methylate in 15 of 35 cases with higher DNA methyltransferase activity whereas only 7 detected to methylate in the remaining 54 cases with lower DNA methyltransferase activity. p16 promoter CpG island methylation likely made p16 expressing silence thus contributed to the tumorigenesis of lung adenocarcinoma. Smoking is likely to promote p16 CpG island methylation or by its effect of the activity and metabolism of DNA methyltransferase 1 (DNMT) on CpG island methylation status.

  20. 唾液腺腺样囊性癌中RASSF1A表达及与启动子甲基化之间的关系%The relationship between RASSF1A expression and promoter methylation in salivary adenoid cystic carcinoma

    Institute of Scientific and Technical Information of China (English)

    李蕾; 夏荣辉; 张春叶; 李江

    2014-01-01

    目的:研究唾液腺腺样囊性癌(adenoid cystic carcinoma,ACC)中RASSF1A表达及其与启动子区甲基化之间的关系.方法:收集167例原发性唾液腺ACC,亚硫酸盐测序聚合酶链反应(bisulfite sequencing polymerase chain reaction,BSP)和甲基化特异性聚合酶链反应(methylation-specific polymerase chain reaction,MSP)方法检测RASSF1A基因启动子区甲基化状况,免疫组织化学方法检测RASSF1A蛋白表达情况.去甲基化药物decitabine处理ACC细胞系SACC-83后,检测处理前、后RASSF1A基因甲基化及表达情况.应用SPSS 18.0软件包对数据进行统计学分析.结果:59/167(35.3%)例病例中检测到RASSF1A基因启动子区甲基化.101/167(60.5%)例病例中RASSF1A蛋白呈低或不表达,66/167(39.5)病例中RASSF1A蛋白呈高表达.存在RASSF1A基因甲基化组,RASSF1A蛋白表达显著低于不存在甲基化组(P=0.012).去甲基化药物decitabine处理ACC细胞系后,RASSF1A mRNA及蛋白水平表达均升高.结论:唾液腺ACC中,启动子区甲基化是RASSF1A基因失活的主要原因,可作为该肿瘤的潜在治疗靶点.

  1. Inheritance and variation of Cytosine methylation in three populus allotriploid populations with different heterozygosity.

    Directory of Open Access Journals (Sweden)

    Yujing Suo

    Full Text Available DNA methylation is an epigenetic mechanism with the potential to regulate gene expression and affect plant phenotypes. Both hybridization and genome doubling may affect the DNA methylation status of newly formed allopolyploid plants. Previous studies demonstrated that changes in cytosine methylation levels and patterns were different among individual hybrid plant, therefore, studies investigating the characteristics of variation in cytosine methylation status must be conducted at the population level to avoid sampling error. In the present study, an F1 hybrid diploid population and three allotriploid populations with different heterozygosity [originating from first-division restitution (FDR, second-division restitution (SDR, and post-meiotic restitution (PMR 2n eggs of the same female parent] were used to investigate cytosine methylation inheritance and variation relative to their common parents using methylation-sensitive amplification polymorphism (MSAP. The variation in cytosine methylation in individuals in each population exhibited substantial differences, confirming the necessity of population epigenetics. The total methylation levels of the diploid population were significantly higher than in the parents, but those of the three allotriploid populations were significantly lower than in the parents, indicating that both hybridization and polyploidization contributed to cytosine methylation variation. The vast majority of methylated status could be inherited from the parents, and the average percentages of non-additive variation were 6.29, 3.27, 5.49 and 5.07% in the diploid, FDR, SDR and PMR progeny populations, respectively. This study lays a foundation for further research on population epigenetics in allopolyploids.

  2. 5-氮杂-2'-脱氧胞苷对 HepG2中 SLIT2甲基化和细胞恶性表型影响的实验研究%EFFECTS OF 5-AZA-2DC ON SLIT2 GENE METHYLATION STATUS AND MALIGNANT PHENOTYPE ON HEPATOMA HEPG2 CELLS

    Institute of Scientific and Technical Information of China (English)

    戴欣; 耿小平; 李晓明; 朱立新; 赵红川; 刘付宝; 王国斌; 赵义军; 黄帆

    2011-01-01

    目的 评估5-氮杂-2'-脱氧胞苷(5-aza-2dc)对人肝癌细胞系 HepG2 中抑癌基因 SLIT2 甲基化状态和细胞增殖、迁移、侵袭和凋亡的影响,探讨SLIT2基因在肝癌中的作用及5-aza-2dc的体外抑制肝癌的效应.方法 实验分为对照组和加药组,分别应用MSP、MTT、划痕实验、侵袭实验、Annexin V-FITC/PI 双染实验检测细胞中 SLIT2 基因的甲基化状态和细胞的增殖、迁移、侵袭、凋亡的变化.结果 5-aza-2dc 处理后,HepG2 细胞株中 SLIT2 基因甲基化程度得到一定程度逆转,并呈现出剂量依赖性;划痕实验48 h后,对照组肝癌细胞明显向划痕的中央迁移,而药物处理组肝癌细胞迁移受到抑制;侵袭实验24 h后,加药组细胞的侵袭能力较对照组降低(P<0.05); Annexin V-FITC/PI双染实验显示加药组细胞凋亡率显著增加(P<0.01).结论 5-aza-2dc对HepG2 细胞有抑制细胞增殖、促进凋亡及抑制细胞迁移和侵袭能力等作用,其中对抑制细胞迁移和侵袭的作用可能是通过逆转 SILT2 基因甲基化、恢复其表达来实现的.%Objactive To evaluate 5-aza-2dc's effect on SLJT2 gene methylation status and cell growth, migration, invasion and apoptosis on hepatoma cell line HepG2, explore the possible effect of SLIT2 gene in hepatocarcinoraa and the depression effect of 5-aza-2dc on hepatocarcinoma in vitro. Methods Two groups were designed in this experiment: control and drug treatment group. The changes of 5-aza-2dc on SLJT2 gene methylation status and the proliferation, migration, invasion and apoptosis of HepG2 cells was detected by MSP, MTT, wound healing assay, cell migration assay and Annexin V-FITC /PI assay, respectively. Results After treated with 5-aza-2dc, the methylation status of SLIT2 gene had been reversed to some extent and showed a dose-dependent; After 48 h in wound healing assay, the control group of HCC cells moved to the center of the scratch, and drug treatment group was

  3. Colorectal carcinomas with KRAS mutation are associated with distinctive morphological and molecular features.

    Science.gov (United States)

    Rosty, Christophe; Young, Joanne P; Walsh, Michael D; Clendenning, Mark; Walters, Rhiannon J; Pearson, Sally; Pavluk, Erika; Nagler, Belinda; Pakenas, David; Jass, Jeremy R; Jenkins, Mark A; Win, Aung Ko; Southey, Melissa C; Parry, Susan; Hopper, John L; Giles, Graham G; Williamson, Elizabeth; English, Dallas R; Buchanan, Daniel D

    2013-06-01

    KRAS-mutated carcinomas comprise 35-40% of all colorectal carcinomas but little is known about their characteristics. The aim of this study was to examine the pathological and molecular features of KRAS-mutated colorectal carcinomas and to compare them with other carcinoma subgroups. KRAS mutation testing was performed in 776 incident tumors from the Melbourne Collaborative Cohort Study. O(6)-methylguanine DNA methyltransferase (MGMT) status was assessed using both immunohistochemistry and MethyLight techniques. Microsatellite instability (MSI) phenotype and BRAF V600E mutation status were derived from earlier studies. Mutation in KRAS codon 12 or codon 13 was present in 28% of colorectal carcinomas. Compared with KRAS wild-type carcinomas, KRAS-mutated carcinomas were more frequently observed in contiguity with a residual polyp (38 vs 21%; Pcarcinomas showed more frequent location in the proximal colon (41 vs 27%; P=0.001), mucinous differentiation (46 vs 25%; Pcarcinomas were distributed in a bimodal pattern along the proximal-distal axis of the colorectum. Compared with male subjects, female subjects were more likely to have KRAS-mutated carcinoma in the transverse colon and descending colon (39 vs 15%; P=0.02). No difference in overall survival was observed in patients according to their tumor KRAS mutation status. In summary, KRAS-mutated carcinomas frequently develop in contiguity with a residual polyp and show molecular features distinct from other colorectal carcinomas, in particular from tumors with neither BRAF nor KRAS mutation.

  4. Comprehensive Molecular Characterization of Papillary Renal Cell Carcinoma

    Science.gov (United States)

    Linehan, W. Marston; Spellman, Paul T.; Ricketts, Christopher J.; Creighton, Chad J.; Fei, Suzanne S.; Davis, Caleb; Wheeler, David A.; Murray, Bradley A.; Schmidt, Laura; Vocke, Cathy D.; Peto, Myron; Al Mamun, Abu Amar M.; Shinbrot, Eve; Sethi, Anurag; Brooks, Samira; Rathmell, W. Kimryn; Brooks, Angela N.; Hoadley, Katherine A.; Robertson, A. Gordon; Brooks, Denise; Bowlby, Reanne; Sadeghi, Sara; Shen, Hui; Weisenberger, Daniel J.; Bootwalla, Moiz; Baylin, Stephen B.; Laird, Peter W.; Cherniack, Andrew D.; Saksena, Gordon; Haake, Scott; Li, Jun; Liang, Han; Lu, Yiling; Mills, Gordon B.; Akbani, Rehan; Leiserson, Mark D.M.; Raphael, Benjamin J.; Anur, Pavana; Bottaro, Donald; Albiges, Laurence; Barnabas, Nandita; Choueiri, Toni K.; Czerniak, Bogdan; Godwin, Andrew K.; Hakimi, A. Ari; Ho, Thai; Hsieh, James; Ittmann, Michael; Kim, William Y.; Krishnan, Bhavani; Merino, Maria J.; Mills Shaw, Kenna R.; Reuter, Victor E.; Reznik, Ed; Shelley, Carl Simon; Shuch, Brian; Signoretti, Sabina; Srinivasan, Ramaprasad; Tamboli, Pheroze; Thomas, George; Tickoo, Satish; Burnett, Kenneth; Crain, Daniel; Gardner, Johanna; Lau, Kevin; Mallery, David; Morris, Scott; Paulauskis, Joseph D.; Penny, Robert J.; Shelton, Candace; Shelton, W. Troy; Sherman, Mark; Thompson, Eric; Yena, Peggy; Avedon, Melissa T.; Bowen, Jay; Gastier-Foster, Julie M.; Gerken, Mark; Leraas, Kristen M.; Lichtenberg, Tara M.; Ramirez, Nilsa C.; Santos, Tracie; Wise, Lisa; Zmuda, Erik; Demchok, John A.; Felau, Ina; Hutter, Carolyn M.; Sheth, Margi; Sofia, Heidi J.; Tarnuzzer, Roy; Wang, Zhining; Yang, Liming; Zenklusen, Jean C.; Zhang, Jiashan (Julia); Ayala, Brenda; Baboud, Julien; Chudamani, Sudha; Liu, Jia; Lolla, Laxmi; Naresh, Rashi; Pihl, Todd; Sun, Qiang; Wan, Yunhu; Wu, Ye; Ally, Adrian; Balasundaram, Miruna; Balu, Saianand; Beroukhim, Rameen; Bodenheimer, Tom; Buhay, Christian; Butterfield, Yaron S.N.; Carlsen, Rebecca; Carter, Scott L.; Chao, Hsu; Chuah, Eric; Clarke, Amanda; Covington, Kyle R.; Dahdouli, Mahmoud; Dewal, Ninad; Dhalla, Noreen; Doddapaneni, HarshaVardhan; Drummond, Jennifer; Gabriel, Stacey B.; Gibbs, Richard A.; Guin, Ranabir; Hale, Walker; Hawes, Alicia; Hayes, D. Neil; Holt, Robert A.; Hoyle, Alan P.; Jefferys, Stuart R.; Jones, Steven J.M.; Jones, Corbin D.; Kalra, Divya; Kovar, Christie; Lewis, Lora; Li, Jie; Ma, Yussanne; Marra, Marco A.; Mayo, Michael; Meng, Shaowu; Meyerson, Matthew; Mieczkowski, Piotr A.; Moore, Richard A.; Morton, Donna; Mose, Lisle E.; Mungall, Andrew J.; Muzny, Donna; Parker, Joel S.; Perou, Charles M.; Roach, Jeffrey; Schein, Jacqueline E.; Schumacher, Steven E.; Shi, Yan; Simons, Janae V.; Sipahimalani, Payal; Skelly, Tara; Soloway, Matthew G.; Sougnez, Carrie; Tam, Angela; Tan, Donghui; Thiessen, Nina; Veluvolu, Umadevi; Wang, Min; Wilkerson, Matthew D.; Wong, Tina; Wu, Junyuan; Xi, Liu; Zhou, Jane; Bedford, Jason; Chen, Fengju; Fu, Yao; Gerstein, Mark; Haussler, David; Kasaian, Katayoon; Lai, Phillip; Ling, Shiyun; Radenbaugh, Amie; Van Den Berg, David; Weinstein, John N.; Zhu, Jingchun; Albert, Monique; Alexopoulou, Iakovina; Andersen, Jeremiah J; Auman, J. Todd; Bartlett, John; Bastacky, Sheldon; Bergsten, Julie; Blute, Michael L.; Boice, Lori; Bollag, Roni J.; Boyd, Jeff; Castle, Erik; Chen, Ying-Bei; Cheville, John C.; Curley, Erin; Davies, Benjamin; DeVolk, April; Dhir, Rajiv; Dike, Laura; Eckman, John; Engel, Jay; Harr, Jodi; Hrebinko, Ronald; Huang, Mei; Huelsenbeck-Dill, Lori; Iacocca, Mary; Jacobs, Bruce; Lobis, Michael; Maranchie, Jodi K.; McMeekin, Scott; Myers, Jerome; Nelson, Joel; Parfitt, Jeremy; Parwani, Anil; Petrelli, Nicholas; Rabeno, Brenda; Roy, Somak; Salner, Andrew L.; Slaton, Joel; Stanton, Melissa; Thompson, R. Houston; Thorne, Leigh; Tucker, Kelinda; Weinberger, Paul M.; Winemiller, Cythnia; Zach, Leigh Anne; Zuna, Rosemary

    2016-01-01

    Background Papillary renal cell carcinoma, accounting for 15% of renal cell carcinoma, is a heterogeneous disease consisting of different types of renal cancer, including tumors with indolent, multifocal presentation and solitary tumors with an aggressive, highly lethal phenotype. Little is known about the genetic basis of sporadic papillary renal cell carcinoma; no effective forms of therapy for advanced disease exist. Methods We performed comprehensive molecular characterization utilizing whole-exome sequencing, copy number, mRNA, microRNA, methylation and proteomic analyses of 161 primary papillary renal cell carcinomas. Results Type 1 and Type 2 papillary renal cell carcinomas were found to be different types of renal cancer characterized by specific genetic alterations, with Type 2 further classified into three individual subgroups based on molecular differences that influenced patient survival. MET alterations were associated with Type 1 tumors, whereas Type 2 tumors were characterized by CDKN2A silencing, SETD2 mutations, TFE3 fusions, and increased expression of the NRF2-ARE pathway. A CpG island methylator phenotype (CIMP) was found in a distinct subset of Type 2 papillary renal cell carcinoma characterized by poor survival and mutation of the fumarate hydratase (FH) gene. Conclusions Type 1 and Type 2 papillary renal cell carcinomas are clinically and biologically distinct. Alterations in the MET pathway are associated with Type 1 and activation of the NRF2-ARE pathway with Type 2; CDKN2A loss and CIMP in Type 2 convey a poor prognosis. Furthermore, Type 2 papillary renal cell carcinoma consists of at least 3 subtypes based upon molecular and phenotypic features. PMID:26536169

  5. Parotid carcinoma

    DEFF Research Database (Denmark)

    Godballe, Christian; Schultz, Joyce H; Krogdahl, Annelise

    2003-01-01

    %) were excluded from the analysis, leaving 75 individuals for the study. RESULTS: Twenty-four percent of patients were classified as T1, 32% as T2, 15% as T3, and 28% as T4. One patient (1%) was not classifiable (TX). Regional lymph node metastases were found in 17 cases (23%). Two of these were......-year recurrence-free survival of the entire study group was 63%, disease-specific survival was 69%, and crude survival was 52%. In univariate analysis, tumor size, histological appearance, T status, stage, the presence of lymph node metastases, distant metastases, pain, and facial nerve dysfunction had...

  6. KRAS gene mutations are more common in colorectal villous adenomas and in situ carcinomas than in carcinomas

    Science.gov (United States)

    Zauber, Peter; Marotta, Stephen; Sabbath-Solitare, Marlene

    2013-01-01

    We have evaluated the frequency of KRAS gene mutations during the critical transition from villous adenoma to colorectal carcinoma to assess whether the adenomas contain a KRAS mutation more frequently than carcinomas. We analyzed sporadic villous and tubulovillous adenomas, in situ carcinomas, and primary colorectal carcinomas from multiple patients. The cancers were further evaluated for mucinous status and microsatellite instability. Standard PCR molecular techniques were used for KRAS and microsatellite analyses. A KRAS mutation was found in 61.9% of 134 adenomas, 67.8% of 84 in situ carcinomas, and just 31.6% of 171 carcinomas. Our study clearly demonstrates that tubulovillous and villous adenomas, as well as both the benign and malignant parts of in situ carcinomas, are statistically more likely to contain a somatic KRAS gene mutation than colorectal carcinomas. This difference is confined to the non-mucinous and the microsatellite stable tumors. Our data support the possibility that non-mucinous and microsatellite stable carcinomas with wild-type KRAS gene may have had a mutation in the KRAS gene during their earlier stages, with the mutation lost during further growth. PMID:23565319

  7. APC2 and CYP1B1 methylation changes in the bone marrow of acute myeloid leukemia patients during chemotherapy

    OpenAIRE

    XIA, YONGMING; Hong, Qingxiao; Chen, Xiaoying; YE, HUADAN; Fang, Lili; Zhou, Annan; GAO, YUTING; Jiang, Danjie; Duan, Shiwei

    2016-01-01

    Aberrant promoter DNA methylation is a major mechanism of leukemogenesis in hematologic malignancies, including acute myeloid leukemia (AML). However, the association between promoter methylation with chemotherapeutic outcomes remains unknown. In the present study, bone marrow samples were collected prior to and following chemotherapy in 30 AML patients. Methylation-specific polymerase chain reaction technology was used to examine the promoter methylation status of adenomatous polyposis col 2...

  8. Epigenetic regulation of Wnt signaling pathway gene SRY-related HMG-box 17 in papillary thyroid carcinoma

    Institute of Scientific and Technical Information of China (English)

    LI Jing-yi; HAN Chao; ZHENG Li-li; GUO Ming-zhou

    2012-01-01

    Background SRY-related HMG-box 17 (SOX17) encodes a member of the SOX (SRY-related HMG-box) family of transcription factors involved in the regulation of embryonic development and in the determination of the cell fate.Recently,it was considered as a tumor suppressor gene to inhibit canonical Wnt/β-catenin signaling pathway in several malignancies.However,the function of SOX17 in thyroid cancer was unknown.Therefore,we investigated the epigenetic changes and the function of SOX17 in thyroid cancer.Methods The methylation status of the promoter region of SOX17 was detected using methylation-specific PCR in 63 papillary thyroid carcinoma (PTC) tissue,10 normal thyroid tissue,and two thyroid cancer cell lines.Semi-quantitative RT-PCR was used to assess mRNA expression of SOX17 before and after 5-aza-2′-deoxycytidine treatment in thyroid cancer cell lines.Expression of SOX17 and β-catenin were detected by immunohistochemistry in PTC and adjacent tissue.Luciferase reporter assay,colony formation,transfection,and Western blotting were employed to analyze the effect of SOX17 on thyroid cancer cell proliferation and the function of SOX17 in the Wnt signal pathway.Results Loss of SOX17 expression was correlated to the promoter region hypermethylation in thyroid cancer cell lines.Re-expression of SOX17 was found in TPC-1 cell line after 5-aza-2′-deoxycytidine treatment.In primary thyroid cancer,60.3% (38/63) were methylated and 39.7% (25/63) unmethylated.But no methylation was found in noncancerous thyroid tissues.Methylation of SOX17 was associated reversely with β-catenin expression in the cytoplasm or nucleus significantly in the PTC (P <0.05).Colony formation was inhibited by re-expression of SOX17 in TPC-1 cells.SOX17 suppressed the Wnt signaling pathway and the HMG domain was essential for this effect.Conclusions SOX17 was frequently methylated in human PTC.Loss of SOX17 expression was induced by promoter region hypermethylation.SOX17 inhibited thyroid

  9. Polyploidy and DNA methylation: new tools available.

    Science.gov (United States)

    Salmon, Armel; Ainouche, Malika L

    2010-01-01

    Most plant species are recent or ancient polyploids (displaying at least one round of genome duplication in their history). Cultivated species (e.g. wheat, cotton, canola, sugarcane, coffee) and invasive species are often relatively recent polyploids, and frequently of hybrid origin (i.e. allopolyploids). Despite the genetic bottleneck occurring during the allopolyploid speciation process, the formation of such species from two divergent lineages leads to fixed heterozygosity decisive to their success. New phenotypes and new niche occupation are usually associated with this mode of speciation, as a result of both genomic rearrangements and gene expression changes of different magnitudes depending on the different polyploid species investigated. These gene expression changes affecting newly formed polyploid species may result from various, interconnected mechanisms, including (i) functional interactions between the homoeologous copies and between their products, that are reunited in the same nucleus and cell; (ii) the fate of duplicated copies, selective pressure on one of the parental copy being released which could lead to gene loss, pseudogenization, or alternatively, to subfunctionalization or neofunctionalization; and (iii) epigenetic landscape changes that in turn affect gene expression. As one of the interrelated processes leading to epigenetic regulation of gene expression, the DNA methylation status of newly formed species appears to be consistently affected following both hybridization and genome doubling. In this issue, Verhoeven et al. have investigated the fate of DNA methylation patterns that could affect naturally occurring new asexual triploid lineages of dandelions. As a result of such a ploidy level change, the authors demonstrate stably transmitted DNA methylation changes leading to unique DNA methylation patterns in each newly formed lineage. Most studies published to date on plant DNA methylation polymorphism were performed using restriction

  10. The Correlation of MGMT Promoter Methylation and Clinicopathological Features in Gastric Cancer: A Systematic Review and Meta-Analysis.

    Science.gov (United States)

    Ding, Yong; Yang, Qihua; Wang, Bojun; Ye, Guoliang; Tong, Xiaoqiong

    2016-01-01

    The silencing of the tumor suppressor gene O-6-methylguanine-DNA methyltransferase (MGMT) by promoter methylation commonly occurs in human cancers. The relationship between MGMT promoter methylation and gastric cancer (GC) remains inconsistent. This study aimed to evaluate the potential value of MGMT promoter methylation in GC patients. Electronic databases were searched to identify eligible studies. The pooled odds ratio (OR) and the corresponding 95% confidence interval (95% CI) were used to evaluate the effects of MGMT methylation on GC risk and clinicopathological characteristics. In total, 31 eligible studies including 2988 GC patients and 2189 nonmalignant controls were involved in meta-analysis. In the pooled analysis, MGMT promoter methylation was significantly associated with GC risk (OR = 3.34, P MGMT methylation showed a trend associated with gender, and methylation is lower in males compared with females (OR = 0.76, 95% CI = 0.56-1.03). We did not find a significant association in relation to tumor types, clinical stage, age status or H. pylori status in cancer (all P > 0.1). MGMT promoter methylation may be correlated with the prognosis of GCs in disease free survival (DFS) or overall survival (OS) for univariate analysis. MGMT promoter methylation may play a crucial role in the carcinogenesis and prognosis of GC. MGMT methylation was not correlated with tumor types, clinical stage, age status, H. pylori status. However, the result of the association of MGMT methylation and gender should be considered with caution.

  11. Biological and clinical significance of NAC1 expression in cervical carcinomas: a comparative study between squamous cell carcinomas and adenocarcinomas/adenosquamous carcinomas.

    Science.gov (United States)

    Yeasmin, Shamima; Nakayama, Kentaro; Rahman, Mohammed Tanjimur; Rahman, Munmun; Ishikawa, Masako; Katagiri, Atsuko; Iida, Kouji; Nakayama, Naomi; Otuski, Yoshiro; Kobayashi, Hiroshi; Nakayama, Satoru; Miyazaki, Kohji

    2012-04-01

    This study examined the biological and clinical significance of NAC1 (nucleus accumbens associated 1) expression in both cervical squamous cell carcinomas and adenocarcinomas/adenosquamous carcinomas. Using immunohistochemistry, the frequency of positive NAC1 expression in adenocarcinomas/adenosquamous carcinomas (31.0%; 18/58) was significantly higher than that in squamous cell carcinomas (16.2%; 12/74) (P = .043). NAC1 gene amplification was identified by fluorescence in situ hybridization in 5 (7.2%) of 69 squamous cell carcinomas. NAC1 amplification was not identified in the adenocarcinomas (0%; 0/58). Positive NAC1 expression was significantly correlated with shorter overall survival in squamous cell carcinomas (P NAC1 expression in squamous cell carcinomas was an independent prognostic factor for overall survival after standard radiotherapy (P = .0003). In contrast to squamous cell carcinomas, positive NAC1 expression did not correlate with shorter overall survival in adenocarcinomas/adenosquamous carcinomas (P = .317). Profound growth inhibition, increased apoptosis, decreased cell proliferation, and decreased cell migration and invasion were observed in silencing RNA-treated cancer cells with NAC1 overexpression compared with cancer cells without NAC1 expression. NAC1 overexpression stimulated proliferation, migration, and invasion in the cervical cancer cell lines TCS and Hela P3, which normally lack NAC1 expression. These findings indicate that NAC1 overexpression is critical to the growth and survival of cervical carcinomas irrespective of histologic type. Furthermore, they suggest that NAC1 silencing RNA-induced phenotypes depend on the expression status of the targeted cell line. Therefore, cervical carcinoma patients with NAC1 expression may benefit from a targeted therapy irrespective of histologic type.

  12. Disclosing bias in bisulfite assay: MethPrimers underestimate high DNA methylation.

    Directory of Open Access Journals (Sweden)

    Andrea Fuso

    Full Text Available Discordant results obtained in bisulfite assays using MethPrimers (PCR primers designed using MethPrimer software or assuming that non-CpGs cytosines are non methylated versus primers insensitive to cytosine methylation lead us to hypothesize a technical bias. We therefore used the two kinds of primers to study different experimental models and methylation statuses. We demonstrated that MethPrimers negatively select hypermethylated DNA sequences in the PCR step of the bisulfite assay, resulting in CpG methylation underestimation and non-CpG methylation masking, failing to evidence differential methylation statuses. We also describe the characteristics of "Methylation-Insensitive Primers" (MIPs, having degenerated bases (G/A to cope with the uncertain C/U conversion. As CpG and non-CpG DNA methylation patterns are largely variable depending on the species, developmental stage, tissue and cell type, a variable extent of the bias is expected. The more the methylome is methylated, the greater is the extent of the bias, with a prevalent effect of non-CpG methylation. These findings suggest a revision of several DNA methylation patterns so far documented and also point out the necessity of applying unbiased analyses to the increasing number of epigenomic studies.

  13. METHYLATION PATTERN OF LRP15 GENE IN LEUKEMIA

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Objective To investigate the methylation status of LRP15 gene in acute leukemia (AL) patients and its role in the tumorigenesis.Methods The methylation of LRP15 promoter and first exon of bone marrow mononuclear cells in 73 patients with AL, 10 with chronic leukemia (CL), 9 with hematological benign diseases, and 20 healthy transplantation donors was analyzed by using methylation specific polymerase chain reaction. The methylation of LRP15 gene promoter and first exon in COS7, K562, and HL60 cell lines was also assayed.Results No LRP15 gene promoter methylation was detected in COS7 cell line. LRP15 gene promoter was methylated in K562 and HL60 cell lines. No deletion of LRP15 gene was detected in all samples. In nearly all French-American-British leukemia subtypes, we found that frequency of LRP15 methylation in adult patients with AL was 71.23%( 52/73 ). There was no detectable methylation in any of the 20 healthy donors and 8 chronic myeloid leukemia patients.The difference in frequency of LRP15 methylation between AL patients and healthy donors or CL patients ( 10. 00%,1/10) was significant (P < 0. 01 ). Hypermethylation of LRP15 gene was found in 57. 14% (16/28) of newly diagnosed AL patients, 83.33% of relapsed AL patients respectively, which was significantly different ( P < 0. 05). We also demonstrated LRP15 methylation in 55.56% (5/9) adults with benign hematological diseases.Conclusions LRP15 methylation changes are common abnormalities in leukemia. LRP15 is postulated to be a tumor suppressor gene.

  14. Correlation of DNA methylation status of imprinted gene H19 ICR with oligozoospermia and asthenozoospermia%父源印记基因H19印记控制区域DNA甲基化与少、弱精子症相关性分析

    Institute of Scientific and Technical Information of China (English)

    李建波; 李博; 梁新新; 王珺; 马夜肥; 张永琦; 刘正; 闵保华; 马旭辉

    2013-01-01

    目的:研究印记基因H19印记控制区域的DNA甲基化程度与男性少、弱精子症的相关性. 方法:通过染色体核型分析和Y染色体微缺失检测排除染色体因素干扰,进一步借助精液常规参数、伊红染色以及精子形态等指标,筛选出18例单一因素少精子症患者(浓度< 15×106/ml,其余指标均正常)和20例单一因素弱精子症患者(前向运动精子百分率<32%,其余指标均正常)用于DNA甲基化检测;提取精子样本全基因组DNA,进行亚硫酸氢盐处理、PCR扩增目的基因片段并测序;通过BIQ Analyzer软件对测序结果进行质控和DNA甲基化程度分析.20例正常生育男性精液标本作为对照组. 结果:与对照组甲基化丢失率[(0.30±0.06)%]相比,少精子症患者的H19印记控制区域的DNA甲基化丢失程度显著增高[(9.19±2.45)%],尤其当精子浓度<3×106/ml时,差异达到极显著水平(P<0.0l).在弱精子症患者中H19印记控制区域的DNA甲基化丢失程度[(0.30±0.07)%]和模式均与对照组无显著差异(P=0.62).进一步重点分析了CTCF6位点的DNA甲基化状态,少精子症患者的DNA甲基化丢失程度[(2.67±0.75)%]显著高于对照组[(0.05±0.03)%]和弱精子症组[(0.03±0.02)%],而后两者之间无显著差异(P=0.35). 结论:H19印记控制区域的DNA甲基化程度的降低与少精子症密切相关,且降低程度与精子浓度呈显著负相关,而与精子活力无关.%Objective:To study the correlation of the DNA methylation status of the imprinted gene H19 imprinting control region (ICR) with oligozoospermia and asthenozoosperrnia.Methods:We eliminated chromosomal abnormality as the cause of male infertility in the subjects by karyotype analysis and detection of Y-chromosome microdeletions,and identified 18 cases of single factorinduced oligozoospermia (sperm concentration < 15 × 106/ml) and 20 cases of single factor-induced asthenozoospermia (progressively motile

  15. Squamoid eccrine ductal carcinoma*

    Science.gov (United States)

    Saraiva, Maria Isabel Ramos; Vieira, Marcella Amaral Horta Barbosa; Portocarrero, Larissa Karine Leite; Fraga, Rafael Cavanellas; Kakizaki, Priscila; Valente, Neusa Yuriko Sakai

    2016-01-01

    Squamoid eccrine ductal carcinoma is an eccrine carcinoma subtype, and only twelve cases have been reported until now. It is a rare tumor and its histopathological diagnosis is difficult. Almost half of patients are misdiagnosed as squamous cell carcinoma by the incisional biopsy. We report the thirteenth case of squamoid eccrine ductal carcinoma. Female patient, 72 years old, in the last 6 months presenting erythematous, keratotic and ulcerated papules on the nose. The incisional biopsy diagnosed squamoid eccrine ductal carcinoma. After excision, histopathology revealed positive margins. A wideningmargins surgery and grafting were performed, which again resulted in positive margins. The patient was then referred for radiotherapy. After 25 sessions, the injury reappeared. After another surgery, although the intraoperative biopsy showed free surgical margins, the product of resection revealed persistent lesion. Distinction between squamoid eccrine ductal carcinoma and squamous cell carcinoma is important because of the more aggressive nature of the first, which requires wider margins surgery to avoid recurrence. PMID:28099603

  16. Squamoid eccrine ductal carcinoma.

    Science.gov (United States)

    Saraiva, Maria Isabel Ramos; Vieira, Marcella Amaral Horta Barbosa; Portocarrero, Larissa Karine Leite; Fraga, Rafael Cavanellas; Kakizaki, Priscila; Valente, Neusa Yuriko Sakai

    2016-01-01

    Squamoid eccrine ductal carcinoma is an eccrine carcinoma subtype, and only twelve cases have been reported until now. It is a rare tumor and its histopathological diagnosis is difficult. Almost half of patients are misdiagnosed as squamous cell carcinoma by the incisional biopsy. We report the thirteenth case of squamoid eccrine ductal carcinoma. Female patient, 72 years old, in the last 6 months presenting erythematous, keratotic and ulcerated papules on the nose. The incisional biopsy diagnosed squamoid eccrine ductal carcinoma. After excision, histopathology revealed positive margins. A wideningmargins surgery and grafting were performed, which again resulted in positive margins. The patient was then referred for radiotherapy. After 25 sessions, the injury reappeared. After another surgery, although the intraoperative biopsy showed free surgical margins, the product of resection revealed persistent lesion. Distinction between squamoid eccrine ductal carcinoma