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1

Targeting Hela cancer cells with avidin-dendrimer  

International Nuclear Information System (INIS)

Dendrimers can target tumor taking advantage of enhanced permeation and retention effect (EPR). In this study,partially acetylated generation 5 (G5) polyamidoamine (PAMAM) dendrimer was conjugated with the active targeting moiety (avidin). The nanoparticles which have passive and active tumor-targeting were radiolabel with 99mTc. The nano-particle conjugated with avidin (avidin-G5-1B4M-99mTc) exhibited much higher cellular uptake into Hela cells than those without avidin conjugate. Using this conjugate, excellent SPECT imaging results were obtained in tumor-bearing nude. This radiolabelling nano-particle has the potential of early stage cancer diagnosis, and cancer therapy. (authors)

2011-01-01

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Amygdalin induces apoptosis in human cervical cancer cell line HeLa cells.  

UK PubMed Central (United Kingdom)

Amygdalin, a naturally occurring substance, has been suggested to be efficacious as an anticancer substance. The effect of amygdalin on cervical cancer cells has never been studied. In this study, we found that the viability of human cervical cancer HeLa cell line was significantly inhibited by amygdalin. 4,6-Diamino-2-phenyl indole (DAPI) staining showed that amygdalin-treated HeLa cells developed typical apoptotic changes. The development of apoptosis in the amygdalin-treated HeLa cells were confirmed by double staining of amygdalin-treated HeLa cells with annexin V-FITC and propidium iodide (PI) along with increase in caspase-3 activity in these cells. Further studies indicated that antiapoptotic protein Bcl-2 was downregulated whereas proapoptotic Bax protein was upregulated in the amygdalin-treated HeLa cells implying involvement of the intrinsic pathway of apoptosis. In vivo, amygdalin administration inhibited the growth of HeLa cell xenografts through a mechanism of apoptosis. The results in the present study suggest that amygdalin may offer a new therapeutic option for patients with cervical cancer.

Chen Y; Ma J; Wang F; Hu J; Cui A; Wei C; Yang Q; Li F

2013-02-01

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Effect of quercetin on radiosensitivity of human uterine cervix cancer HeLa cells  

International Nuclear Information System (INIS)

[en] In order to investigate the effects of Quercetin on radiosensitivity of human Uterine Cervix Cancer HeLa cells, MTT assay and clonogenic assay were performed to evaluate the cytotoxicity of Quercetin on the cells. Clonogenic assay was used to observe its effects on the radiosensitivity of the cells. MTT result shows that the inhibition of Quercetin on the cells is in the dose-dependent and time-dependent. And the clonogenic assay result shows that the effect of Quercetin on HeLa cells can be divided into two parts, one for the inhibition of HeLa cells and another for the induction of HeLa cell death. The other clonogenic assay result also shows Quercetin can decrease clonogenic survival rate of HeLa cells exposed to X rays. The study shows Quercetin might enhance the radiosensitivity of the HeLa cell line. And it may provide a useful evaluation to combination of ionizing radiation and Quercetin for cancer patients. (authors)

2009-01-01

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Oral cancer overexpressed 1 (ORAOV1) regulates cell cycle and apoptosis in cervical cancer HeLa cells  

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Full Text Available Abstract Background Oral Cancer Overexpressed 1 (ORAOV1) is a candidate protooncogene locating on 11q13. Recent studies show that ORAOV1 acts as a primary driving force behind 11q13 gene amplification and plays a functional role in the tumorigenesis in a variety of human squamous cell carcinomas (SCCs). According to the results of molecular cytogenetic methods, 11q13 was characterized to be a high-level and recurrent amplification chromosomal site in cervical cancers. Up till now, the role of ORAOV1 in cervical cancer is unknown. The purpose of this study is to elucidate the function of ORAOV1 in cervical cancer cell growth by studying its roles in HeLa cells using small interfering RNA. Results Functional analyses revealed that ORAOV1 was involved in the regulation of HeLa cell growth through its effect on cell cycle and apoptosis. Silence of ORAOV1 in HeLa cells downregulated the expression of Cyclin A, Cyclin B1 and Cdc2, and led to a distinct S cell cycle arrest. Moreover, knockdown of ORAOV1 expression activated both extrinsic and intrinsic apoptotic pathways and led to apoptosis in HeLa cells through its effect on the expression of several apoptosis related proteins such as P53, Bcl-2, Caspase-3, Caspase-8, Caspase-9 and cytochrome c. Interestingly, the expression of Cyclin D1, a pivotal gene for cervical cancer tumorigenesis, was also found to be reduced in ORAOV1 silenced HeLa cells. Conclusion Our findings indicate that ORAOV1 has an important role in regulating cell growth of cervical cancer HeLa cells through regulating the cell cycle and apoptosis. Thus, it may be a crucial protooncogene and a novel candidate therapeutic target for cervical cancer.

Jiang Lu; Zeng Xin; Wang Zhi; Ji Ning; Zhou Yu; Liu Xianting; Chen Qianming

2010-01-01

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[Inhibition of Jumi extraction on growth of human cervical cancer cell line HeLa].  

UK PubMed Central (United Kingdom)

OBJECTIVE: To explore the inhibition of Jumi (traditional Chinese medicine) extraction on the growth of human cervical cancer cell line HeLa. METHODS: Nude mouse model of human cervical cancer HeLa cell transplantation was established. The nude mice bearing cancer were randomly divided into control group and Jumi treated groups with different concentration (0.001, 0.002, 0.005, 0.01 mg/ml). The growth of cervical cancer cell in experimental mice were measured. Cultured HeLa cells were incubated in culture media with or without Jumi extract for 48 hours. Cell proliferation rate, cell apoptosis, caspase-3/7 and caspase-6 activity were determined by MTT colorimetric assay, flow cytometry analysis and spectrophotometric detection, respectively. RESULTS: With the increase of the concentration of Jumi extract, tumor-bearing mice tumor inhibition rate gradually increased. The proliferation of cultured HeLa cells were significantly inhibited by Jumi extract in a dose-dependent manner. IC50 was 0.004 mg/ml. Apoptosis rates in the cells treated with Jumi extract were higher than those of the control group. Compared with the control group, except for lower Jumi treated group (0.001 mg/ml), caspase-3/7 and caspase-6 activity were significantly increased in the all Jumi treated groups. CONCLUSION: Jumi extract can inhibit the proliferation of human cervical cancer cell line HeLa in vitro in a dose-dependent manner and promote cell apoptosis through caspase-3, caspase-7 and caspase-6 pathway.

Kuang W; Chen HL; Jiang JP

2013-05-01

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The haplotype-resolved genome and epigenome of the aneuploid HeLa cancer cell line.  

UK PubMed Central (United Kingdom)

The HeLa cell line was established in 1951 from cervical cancer cells taken from a patient, Henrietta Lacks. This was the first successful attempt to immortalize human-derived cells in vitro. The robust growth and unrestricted distribution of HeLa cells resulted in its broad adoption--both intentionally and through widespread cross-contamination--and for the past 60?years it has served a role analogous to that of a model organism. The cumulative impact of the HeLa cell line on research is demonstrated by its occurrence in more than 74,000 PubMed abstracts (approximately 0.3%). The genomic architecture of HeLa remains largely unexplored beyond its karyotype, partly because like many cancers, its extensive aneuploidy renders such analyses challenging. We carried out haplotype-resolved whole-genome sequencing of the HeLa CCL-2 strain, examined point- and indel-mutation variations, mapped copy-number variations and loss of heterozygosity regions, and phased variants across full chromosome arms. We also investigated variation and copy-number profiles for HeLa S3 and eight additional strains. We find that HeLa is relatively stable in terms of point variation, with few new mutations accumulating after early passaging. Haplotype resolution facilitated reconstruction of an amplified, highly rearranged region of chromosome 8q24.21 at which integration of the human papilloma virus type 18 (HPV-18) genome occurred and that is likely to be the event that initiated tumorigenesis. We combined these maps with RNA-seq and ENCODE Project data sets to phase the HeLa epigenome. This revealed strong, haplotype-specific activation of the proto-oncogene MYC by the integrated HPV-18 genome approximately 500?kilobases upstream, and enabled global analyses of the relationship between gene dosage and expression. These data provide an extensively phased, high-quality reference genome for past and future experiments relying on HeLa, and demonstrate the value of haplotype resolution for characterizing cancer genomes and epigenomes.

Adey A; Burton JN; Kitzman JO; Hiatt JB; Lewis AP; Martin BK; Qiu R; Lee C; Shendure J

2013-08-01

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Knock-down of NDRG2 sensitizes cervical cancer Hela cells to cisplatin through suppressing Bcl-2 expression  

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Full Text Available Abstract Background NDRG2, a member of N-Myc downstream regulated gene family, plays some roles in cellular stress, cell differentiation and tumor suppression. We have found that NDRG2 expression in cervical cancer Hela cells increases significantly upon stimulation with cisplatin, the most popular chemotherapeutic agent currently used for the treatment of advanced cervical cancer. This interesting phenomenon drove us to evaluate the role of NDRG2 in chemosensitivity of Hela cells. Methods In the present study, RNA interference was employed to down-regulate NDRG2 expression in Hela cells. RT-PCR and Western blot were used to detect expression of NDRG2, Bcl-2 and Bax in cancer cells. Real-time PCR was applied to detect miR-15b and miR-16 expression levels. Drug sensitivity was determined with MTT assay. Cell cloning efficiency was evaluated by Colony-forming assay. Apoptotic cells were detected with annexin V staining and flow cytometry. Results In vitro drug sensitivity assay revealed that suppression of NDRG2 could sensitize Hela cells to cisplatin. Down-regulation of NDRG2 didn’t influence the colony-forming ability but promoted cisplatin-induced apoptosis of Hela cells. Inhibition of NDRG2 in Hela cells was accompanied by decreased Bcl-2 protein level. However, Bcl-2 mRNA level was not changed in Hela cells with down-regulation of NDRG2. Further study indicated that miR-15b and miR-16, two microRNAs targetting Bcl-2, were significantly up-regulated in NDRG2-suppressed Hela cells. Conclusions These data suggested that down-regulation of NDRG2 could enhance sensitivity of Hela cells to cisplatin through inhibiting Bcl-2 protein expression, which might be mediated by up-regulating miR-15b and miR-16.

Liu Junye; Yang Le; Zhang Jian; Zhang Jing; Chen Yongbin; Li Kangchu; Li Yurong; Li Yan; Yao Libo; Guo Guozhen

2012-01-01

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Valproic acid inhibits the growth of HeLa cervical cancer cells via caspase-dependent apoptosis.  

UK PubMed Central (United Kingdom)

Valproic acid (VPA) as a histone deacetylase (HDAC) inhibitor has an anticancer effect. In the present study, we evaluated the effects of VPA on the growth and death of HeLa cervical cancer cells in relation to reactive oxygen species (ROS) and glutathione (GSH). Dose- and time-dependent growth inhibition was observed in HeLa cells with an IC50 of approximately 10 mM at 24 h. DNA flow cytometric analysis indicated that 10 mM VPA induced a G2/M phase arrest of the cell cycle. This agent also induced apoptosis, which was accompanied by the cleavage of PARP, the activation of caspase-3, -8 and -9, and the loss of mitochondrial membrane potential (MMP; ??m). All the tested caspase inhibitors significantly prevented HeLa apoptotic cell death induced by VPA, whereas TNF-? intensified the apoptotic cell death. With respect to ROS and GSH levels, VPA increased ROS levels and induced GSH depletion. However, N-acetyl cysteine (NAC; an antioxidant) and L-buthionine sulfoximine (BSO; a GSH synthesis inhibitor) did not significantly affect cell death in VPA-treated HeLa cells. In conclusion, VPA inhibits the growth of HeLa cervical cancer cells via caspase-dependent apoptosis and the growth inhibition is independent of ROS and GSH level changes.

Han BR; You BR; Park WH

2013-09-01

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Anticancer Activity of Natural Compound (Zerumbone) Extracted from Zingiber zerumbet in Human HeLa Cervical Cancer Cells  

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Full Text Available A natural compound, zerumbone was extracted, isolated and purified from the rhizomes of edible plant Zingiber zerumbet using methanol extraction and Column Chromatography (CC) method. The isolated and purified zerumbone crystals were subjected to High Performance Liquid Chromatography (HPLC), Liquid Chromatography Mass Spectrometry (LCMS) and 13C NMR and 1H NMR analysis to confirm the purity, molecular weight and molecular structure. The study investigated the purified zerumbone crystals for its anti-cancer properties on human cervical cancer cell line (HeLa). Cisplatin, was used as a positive control in this study. The cytotoxicity of zerumbone and cisplatin were investigated using the MTT assay and caspases-3 was estimated with colorimetric assay in zerumbone treated HeLa cells. Morphological analysis showed that there were changes observed on HeLa cancer cells after treatment with zerumbone and cisplatin. The MTT assay results demonstrated that the IC50 value ( ± SEM) of zerumbone was determined to be 11.3 ?M (2.5 ?g mL-1) whilst the IC50 value of cisplatin was at 7.5 ?M (1.6 ?g mL-1). Prominent growth retardation was identified to the HeLa cancer cells, after treatment with both compounds, while caspase-3 was observed to be significantly increased in zerumbone treated cells as compared to untreated control cells. This study showed promising avenues towards zerumbone to be developed as a new chemo-natural drug for treatment of cervical cancer.

A.B.H. Abdul; A.S. Al-Zubairi; N.D. Tailan; S.I.A. Wahab; Z.N.M. Zain; S. Ruslay; M.M. Syam

2008-01-01

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Growth inhibition and induction of apoptosis in human cancerous HeLa cells by Maytenus procumbens.  

UK PubMed Central (United Kingdom)

The possible biochemical activities of the acetonic/ethanolic extract of the leaves of Maytenus procumbens (L.M.P), and its isolated compounds were investigated in the present study. In cytotoxicity assay, L.M.P showed IC(50) of 68.79, 51.22, 78.49, 76.59, and 76.64?g/ml on Caco-2, HeLa, HT29, NIH3T3, and T47D cells, respectively. Bioassay guided fractionation led to the isolation and identification of a new triterpene: '30-hydroxy-11?-methoxy-18?-olean-12-en-3-one' (HMO) in addition to a known terpenoid: 'asiatic acid' (AA). HMO exhibited the most cytotoxicity against HeLa cells and was further investigated for its ability to induce apoptosis in HeLa cells. HMO induced apoptosis up to 20.41% in HeLa cells versus control group (0.40%). Antioxidant/oxidative properties of L.M.P and HMO were investigated using extracellular (DPPH), and intracellular (ROS) assays. Experimental samples represented a time and concentration-dependent formation of ROS in Hela cells. Generation of ROS seems one of the mechanisms by which HMO induces apoptosis in Hela cells. Conclusion is that the active components in L.M.P might serve as a mediator of the ROS scavenging system and have the potential to act as prooxidant or antioxidant depending on the biological environment of the cells.

Momtaz S; Hussein AA; Ostad SN; Abdollahi M; Lall N

2013-01-01

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Radiation enhancing effects with the combination of sanazole and irinotecan in hypoxic HeLa human cervical cancer cell line.  

UK PubMed Central (United Kingdom)

Purpose: This study was conducted to determine the synergistic radiation sensitizing effects of the combination of sanazole and irinotecan in hypoxic cervical cancer HeLa human tumor cell line. Methods: The 3-(4,5 dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay was used to evaluate the number of surviving cells. Cell cycle was determined by flow cytometry. Surviving cell fractions were determined by the standard in vitro colony formation assay. Results: The MTT assay showed that the presence of irinotecan with or without sanazole reduced significantly the cells' viability. Flow cytometry demonstrated that the combination of sanazole and irinotecan led to more HeLa cells blocked in G2 phase. Cell colony formation assay indicated that the radiosensitivity of hypoxic HeLa cells was enhanced by sanazole and/or irinotecan. Conclusion: This study showed that the radiation enhancing effects produced by the combination sanazole and irinotecan was significant in hypoxic HeLa cells, which were arrested in the G2 phase of the cell cycle. This study may provide a new combination modality of radiosensitizers in the radiotherapy of cervical cancer.

Zeng YC; Yu L; Xiao YP; Xin Y; Wang NQ; Zhang XY; Zhao L

2013-07-01

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Combination of phenylpropanoids with 5-fluorouracil as anti-cancer agents against human cervical cancer (HeLa) cell line.  

UK PubMed Central (United Kingdom)

Combination therapy is the most effective treatment strategy in cancer to overcome drug toxicity and drug induced resistance. The effect of eight phenylpropanoids in combination with 5-fluorouracil against the cervical cancer cells (HeLa) is reported here. The cytotoxic activity of these phenylpropanoids against HeLa cells is in the order of eugenol>ferulic>cinnamic>caffeic>chlorogenic>p-coumaric>3,4-dimethoxycinnamic>2,4,5-trimethoxycinnamic acids. Eugenol, ferulic and caffeic acids interacted synergistically with the drug, in bringing about a reduction in the amount of the latter. Flow cytometry results indicated that the combination of eugenol and 5-fluorouracil increased the number of cells in the S and G2/M phases when compared to treatment with the individual compounds alone. This indicates that they possess different cell cycle targets and induce apoptosis in the cancer cells. In vitro hemolytic activity of phenylpropanoids on human erythrocytes showed that the compounds possessed minimum amount of hemolytic activity, indicating that they can be used as drugs without causing adverse toxicity. 3D-quantitative structure activity relationship studies indicate the importance of electrostatic region near the substitutions present in the benzene ring and near the double bond of the compounds for anticancer and hemolytic activities, respectively. The models derived had good statistical predictive capability.

Hemaiswarya S; Doble M

2013-01-01

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Apoptosis induced by yessotoxins in Hela human cervical cancer cells in vitro.  

UK PubMed Central (United Kingdom)

To investigate the apoptotic effects of an extract of red-tide algae, Protoceratium reticulatum, on HeLa cells, we used liquid chromatography-tandem mass spectrometry, optical microscopy, Hoechst 33342/propidine iodide staining, and DNA gel electrophoresis to analyze its constituents and toxicity, as well as rhodamine 123 staining to investigate changes in mitochondrial membrane potential. Analysis showed that the P. reticulatum extract contained the yessotoxins (YTXs) homo-YTX, 45-OH-YTX and 45-OH-homo-YTX. The results indicated that P. reticulatum extract negatively influences HeLa cells and induces their apoptosis.

Pang M; Gao CL; Wu ZX; Lv N; Wang ZL; Tang XX; Qu P

2010-07-01

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Extracellular Caspase-8 Dependent Apoptosis on HeLa Cancer Cells and MRC-5 Normal Cells by ICD-85 (Venom Derived Peptides)  

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Full Text Available Abstract Background: Our previous studies revealed an inhibitory effect of ICD-85 (venom derived peptides) on MDA-MB231 and HL-60 cell lines, through induction of apoptosis. The purpose of this study was to investigate apoptosis-induced mechanism on HeLa and MRC-5 cells by ICD-85 through activation of caspase-8.Methods: Cell viability, cytosolic enzyme Lactate Dehydrogenase (LDH) and cell morphology were assessed under unexposed and ICD-85 exposed conditions.Caspase-8 activity was assayed by caspase-8 colorimetric assay Kit. Results: The results show that Inhibitory Concentration 50% (IC50) value of ICD-85 for HeLa cells at 24 h was estimated and found to be 25.32±2.15µg/ml. Furthermore, treatment of HeLa cells with ICD-85 at concentrations of 1.6×10 and 2.6×10µg/ml did not significantly increase LDH release. Morphological changes in HeLa cells on treatment with ICD-85 compared with untreated HeLa cells consistent with an apoptotic mechanism of cell death, such as cell shrinkage which finally results in the generation of apoptotic bodies. However, when MRC-5 cells were exposed to ICD-85, no significant changes in cell morphology and LDH were observed at concentrations below 2.6×10µg/ml. Also, the apoptosis-induction mechanism by ICD-85 on HeLa cells was found through activation of caspase-8 and the activity of caspase-8 in HeLa cells was 1.5 folds more than its activity on MRC-5 cells. Conclusion: Therefore, the apoptosis-induced mechanisms by ICD-85 are through activation of caspase-8 and concerning the least cytotoxic effect on MRC-5 cells, ICD-85 may be used as anticancer compound to inhibit growth of cancer cells.

Abbas Zare Mirakabadi; Ali Sarzaeem

2012-01-01

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[Molecular mechanism of ophiopogonin B induced cellular autophagy of human cervical cancer HeLa cells].  

UK PubMed Central (United Kingdom)

This study is to investigate the antitumor activity of ophiopogonin B (OP-B). MTT assay, flow cytometric analysis, acridine orange staining, Lyso-Tracker Red staining and HeLa-GFP-LC3 transfect cells assay were used to detect the proliferation activity, apoptosis and autophagy of HeLa cells. The results showed that OP-B exerted potent antiproliferative activity on HeLa cells, the cell growth inhibition effect of OP-B was not due to apoptosis and OP-B could induce autophagy of HeLa cells. OP-B also induced the protein expression up-regulation of Beclin-1 and promoted LC3 I transformation LC3 II, which were representative proteins of autophagy. Furthermore, 3-MA, an inhibitor of autophagy, not only inhibited OP-B-mediated autophagy but also almost completely reversed the antiproliferative effect of OP-B, suggesting that the growth inhibition effect of OP-B was autophagy dependent. Western blotting demonstrated that OP-B inhibited the phosphorylation of Akt and its' downstream vital protein, such as mTOR and p70S6K. In addition, OP-B also induced the protein expression up-regulation of PTEN, which is a negative regulation protein for Akt/mTOR signaling pathway. However, OP-B did not affect the protein expression of total Akt. Collectively, the antitumor effects of OP-B were autophagy-dependent via repression Akt/mTOR signaling pathway. Therefore, OP-B is a prospective inhibitor of Akt/mTOR and may be used as an alternative compound to treat cervical carcinoma.

Xu QJ; Hou LL; Hu GQ; Xie SQ

2013-06-01

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Nogo-B promotes the epithelial-mesenchymal transition in HeLa cervical cancer cells via Fibulin-5.  

UK PubMed Central (United Kingdom)

Cervical cancer is a common malignancy in women worldwide, and the occurrence of invasion and metastasis is the major cause for most cancer-related deaths. Epithelial-mesenchymal transition (EMT) has been implicated in the metastasis of primary tumors and provides molecular mechanisms for cervical cancer metastasis. We previously reported that Nogo-B mediates cell motility by binding Fibulin-5. Herein, we show that the increased expression of Nogo-B is correlated with the degree of cervical cancer metastasis. In HeLa cervical cancer cells, overexpression of Nogo-B induces the EMT and promotes cell migration and invasion, while inhibiting cell adhesion. Furthermore, we found that Nogo-B accumulates and co-localizes with Fibulin-5 in pseudopods, and the downstream effects of overexpression of Nogo-B on cell motility could be partially abolished by RNA interference against Fibulin-5. These results suggest that Nogo-B functions as an inducer of cervical cancer metastasis and that this effect is mediated, at least in part, through Fibulin-5.

Xiao W; Zhou S; Xu H; Li H; He G; Liu Y; Qi Y

2013-01-01

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Selective suppression of cervical cancer Hela cells by 2-O-?-D-glucopyranosyl-L-ascorbic acid isolated from the fruit of Lycium barbarum L.  

Science.gov (United States)

Lycium barbarum fruit has been used as a Chinese traditional medicine and dietary supplement for centuries. 2-O-?-D-Glucopyranosyl-L-ascorbic acid (AA-2?G), a novel stable vitamin C analog, is one of the main biologically active components of the fruit. In this report, we investigated the cytotoxic and antiproliferative effect of AA-2?G against cancer cells in vitro and identified the proteins with significantly differential expression in the cervical cancer cells (Hela) cultured in the presence of AA-2?G proteomic analysis. Our results demonstrated that the cytotoxic and antiproliferative activity of AA-2?G on cancer cell lines were in a cell type-, time-, and dose-dependent manner. Similar to vitamin C, the AA-2?G selectively induced cell death repressed the proliferation of Hela cells by the mechanism of cell apoptosis and cell cycle arrest induced by AA-2?G through a mechanism of stabilizing p53 protein. However, the biological activity of inhibition of cell proliferation in other malignant cancer cell lines or primary cells were varied, as demonstrated by either moderate inhibition or slight promotion following treatment with AA-2?G. Comparative analysis of the proteomic profiles and immunoblot analysis identified 15 proteins associated with repressing cell apoptosis and/or stimulating cell proliferation in Hela cells that were downregulated in the presence of AA-2?G or vitamin C. These data indicate that a mechanism of the AA-2?G and vitamin C mediated antitumor activity by downregulating the expression of proteins involved in cell apoptosis and proliferation and consequently inducing Hela cell apoptosis and cell cycle arrest, suggesting that AA-2?G and vitamin C may share a similar mechanism of inducing Hela cell apoptosis. These results also suggest that the L. barbarum fruit may be a potential dietary supplement and anticancer agent aimed at the prevention and treatment of cervical cancer. PMID:20717715

Zhang, Zhiping; Liu, Xiaoming; Wu, Tao; Liu, Junhong; Zhang, Xu; Yang, Xueyun; Goodheart, Michael J; Engelhardt, John F; Wang, Yujiong

2010-08-19

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Selective suppression of cervical cancer Hela cells by 2-O-?-D-glucopyranosyl-L-ascorbic acid isolated from the fruit of Lycium barbarum L.  

UK PubMed Central (United Kingdom)

Lycium barbarum fruit has been used as a Chinese traditional medicine and dietary supplement for centuries. 2-O-?-D-Glucopyranosyl-L-ascorbic acid (AA-2?G), a novel stable vitamin C analog, is one of the main biologically active components of the fruit. In this report, we investigated the cytotoxic and antiproliferative effect of AA-2?G against cancer cells in vitro and identified the proteins with significantly differential expression in the cervical cancer cells (Hela) cultured in the presence of AA-2?G proteomic analysis. Our results demonstrated that the cytotoxic and antiproliferative activity of AA-2?G on cancer cell lines were in a cell type-, time-, and dose-dependent manner. Similar to vitamin C, the AA-2?G selectively induced cell death repressed the proliferation of Hela cells by the mechanism of cell apoptosis and cell cycle arrest induced by AA-2?G through a mechanism of stabilizing p53 protein. However, the biological activity of inhibition of cell proliferation in other malignant cancer cell lines or primary cells were varied, as demonstrated by either moderate inhibition or slight promotion following treatment with AA-2?G. Comparative analysis of the proteomic profiles and immunoblot analysis identified 15 proteins associated with repressing cell apoptosis and/or stimulating cell proliferation in Hela cells that were downregulated in the presence of AA-2?G or vitamin C. These data indicate that a mechanism of the AA-2?G and vitamin C mediated antitumor activity by downregulating the expression of proteins involved in cell apoptosis and proliferation and consequently inducing Hela cell apoptosis and cell cycle arrest, suggesting that AA-2?G and vitamin C may share a similar mechanism of inducing Hela cell apoptosis. These results also suggest that the L. barbarum fruit may be a potential dietary supplement and anticancer agent aimed at the prevention and treatment of cervical cancer.

Zhang Z; Liu X; Wu T; Liu J; Zhang X; Yang X; Goodheart MJ; Engelhardt JF; Wang Y

2011-04-01

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Internalization pathways of anisotropic disc-shaped zeolite L nanocrystals with different surface properties in HeLa cancer cells.  

UK PubMed Central (United Kingdom)

Information about the mechanisms underlying the interactions of nanoparticles with living cells is crucial for their medical application and also provides indications of the putative toxicity of such materials. Here the uptake and intracellular delivery of disc-shaped zeolite L nanocrystals as porous aminosilicates with well-defined crystal structure, uncoated as well as with COOH-, NH2 -, polyethyleneglycol (PEG)- and polyallylamine hydrochloride (PAH) surface coatings are reported. HeLa cells are used as a model system to demonstrate the relation between these particles and cancer cells. Interactions are studied in terms of their fates under diverse in vitro cell culture conditions. Differently charged coatings demonstrated dissimilar behavior in terms of agglomeration in media, serum protein adsorption, nanoparticle cytotoxicity and cell internalization. It is also found that functionalized disc-shaped zeolite L particles enter the cancer cells via different, partly not yet characterized, pathways. These in vitro results provide additional insight about low-aspect ratio anisotropic nanoparticle interactions with cancer cells and demonstrate the possibility to manipulate the interactions of nanoparticles and cells by surface coating for the use of nanoparticles in medical applications.

Li Z; Hüve J; Krampe C; Luppi G; Tsotsalas M; Klingauf J; De Cola L; Riehemann K

2013-05-01

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Internalization pathways of anisotropic disc-shaped zeolite L nanocrystals with different surface properties in HeLa cancer cells.  

Science.gov (United States)

Information about the mechanisms underlying the interactions of nanoparticles with living cells is crucial for their medical application and also provides indications of the putative toxicity of such materials. Here the uptake and intracellular delivery of disc-shaped zeolite L nanocrystals as porous aminosilicates with well-defined crystal structure, uncoated as well as with COOH-, NH2 -, polyethyleneglycol (PEG)- and polyallylamine hydrochloride (PAH) surface coatings are reported. HeLa cells are used as a model system to demonstrate the relation between these particles and cancer cells. Interactions are studied in terms of their fates under diverse in vitro cell culture conditions. Differently charged coatings demonstrated dissimilar behavior in terms of agglomeration in media, serum protein adsorption, nanoparticle cytotoxicity and cell internalization. It is also found that functionalized disc-shaped zeolite L particles enter the cancer cells via different, partly not yet characterized, pathways. These in vitro results provide additional insight about low-aspect ratio anisotropic nanoparticle interactions with cancer cells and demonstrate the possibility to manipulate the interactions of nanoparticles and cells by surface coating for the use of nanoparticles in medical applications. PMID:23335435

Li, Zhen; Hüve, Jana; Krampe, Christina; Luppi, Gianluigi; Tsotsalas, Manuel; Klingauf, Jürgen; De Cola, Luisa; Riehemann, Kristina

2013-01-18

 
 
 
 
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Emodin induces apoptosis of human cervical cancer hela cells via intrinsic mitochondrial and extrinsic death receptor pathway.  

UK PubMed Central (United Kingdom)

BACKGROUND: Emodin is a natural anthraquinone derivative isolated from the Rheum palmatum L. Aim: The aim of the present study was to investigate the effect of emodin on the apoptosis of the human cervical cancer line HeLa and to identify the mechanisms involved. METHODS: Relative cell viability was assessed by MTT assay after treatment with emodin. Cell apoptosis was detected with TUNEL, Hoechst 33342 staining and quantified with flow cytometry using annexin FITC-PI staining. RESULTS: The percentage of apoptotic cells was 0.8, 8.2, 22.1, and 43.7%, respectively. The mRNA levels of Caspase-9, -8 and -3 detected by Real-time PCR after treatment with emodin were significantly increased. Emodin increased the protein levels of Cytochome c, Apaf-1, Fas, FasL, and FADD but decreased the protein levels of Pro-caspase-9, Pro-caspase-8 and Pro-caspase-3. CONCLUSION: We conclude that the emodin inhibited HeLa proliferation by inducing apoptosis through the intrinsic mitochondrial and extrinsic death receptor pathways.

Yaoxian W; Hui Y; Yunyan Z; Yanqin L; Xin G; Xiaoke W

2013-01-01

22

Cytotoxicity of neoplastic drugs Gefitinib, Cisplatin, 5-FU, Gemcitabine, and Vinorelbine on human cervical cancer cells (HeLa)  

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Full Text Available A recent study reports that more than 99% of cases of cervical cancer worldwide contain HPV DNA. Hence,treatment options for cervical cancers are difficult due to multiplicity of the disease. Chemotherapy uses strong anticancerchemicals to kill cancer cells but to kill the viruses clinicians administer combination of drugs or higher dosesof chemotherapy to control advanced cervical cancer and this practice causes severe side effects. Numerous cancerpatients fail standard chemotherapy or develop resistance to chemotherapy during the course of treatment. Hencethe aim of this investigation was to determine the chemo sensitivity of the five commonly used neoplastic drugs suchas Geftinib, Cisplatin, 5-FU, Gemcitabine and Vinorelbine in vitro, and compare its toxicity on cervical cancer cells(HeLa) using lymphocytes (nucleated cells) as controls. Cytotoxicity in vitro was determined using the MTT assay;LC50 for all the drugs was calculated by regression equation. The morphological change of cells was recorded usingInverted Microscopy. DNA damage studies by comet assay determined the extent of single strand breaks in the DNAand these results were statistically determined using Standard deviation and compared with various treatments incancer cells (HeLa) and control cells. All the results were statistically analyzed and recorded. From these studies wecould determine that cisplatin was the most toxic drug and vinorelbine was least toxic. The order of toxicity (LC50) ofthe neoplastic drugs was Cisplatin (13?M) > Gefitinib (20?M) > Gemcitabine (35?M) > 5-FU (40?M) > Vinorelbine(48?M). Further, we could determine the toxicity of the combination of drugs using sub-lethal doses of each drug.

M Ahmed

2011-01-01

23

Fisetin induces apoptosis in human cervical cancer HeLa cells through ERK1/2-mediated activation of caspase-8-/caspase-3-dependent pathway.  

UK PubMed Central (United Kingdom)

Fisetin is a naturally occurring flavonoid that has been reported to inhibit the proliferation and to induce apoptotic cell death in several tumor cells. However, the apoptosis-inducing effect of fisetin on tumor cell lines was investigated besides HeLa cells. In this study, we found that fisetin induced apoptosis of HeLa cells in a dose- and time-dependent manner, as evidenced by nuclear staining of 4'-6-Diamidino-2-phenylindole (DAPI), flow cytometry assay, and Annexin-V/PI double-labeling. In addition, fisetin triggered the activations of caspases-3 and -8 and the cleavages of poly (ADP-ribose) polymerase, resulting in apoptosis induction. Moreover, treatment of HeLa cells with fisetin induced a sustained activation of the phosphorylation of ERK1/2, and inhibition of ERK1/2 by PD98059 (MEK1/2 inhibitor) or transfection with the mutant ERK1/2 expression vector significantly abolished the fisetin-induced apoptosis through the activation of caspase-8/-3 pathway. The in vivo xenograft mice experiments revealed that fisetin significantly reduced tumor growth in mice with HeLa tumor xenografts. In conclusion, our results indicated that fisetin exhibited anti-cancer effect and induced apoptosis in HeLa cell lines both in vitro and in vivo.

Ying TH; Yang SF; Tsai SJ; Hsieh SC; Huang YC; Bau DT; Hsieh YH

2012-02-01

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Essential role of autophagy in fucoxanthin-induced cytotoxicity to human epithelial cervical cancer HeLa cells.  

UK PubMed Central (United Kingdom)

Aim:To investigate the effects and the molecular mechanisms of fucoxanthin, a major carotenoid found in edible seaweed, on HeLa cells.Methods:The cytotoxicity of fucoxanthin was evaluated using MTT assay. Cell cycle and apoptosis were evaluated using flow cytometric analysis. Autophagy was detected with acridine orange staining and transient transfection of the GFP-LC3 plasmid into the cells. Protein expression was detected with Western blotting.Results:Treatment of HeLa cells with fucoxanthin (10-80 ?mol/L) for 48 h caused dose-dependent cytotoxicity with an IC50 value of 55.1±7.6 ?mol/L. Fucoxanthin (10, 20, and 40 ?mol/L) dose-dependently induced G0/G1 arrest, but did not change the apoptosis of HeLa cells. The same concentrations of fucoxanthin dose-dependently increased the protein expression of LC3 II (the autophagosome marker) and Beclin 1 (the initiation factor for autophagosome formation) in HeLa cells. Moreover, fucoxanthin dose-dependently decreased the levels of phosphorylated Akt and its downstream proteins p53, p70S6K, and mTOR, and increases the expression of PTEN in HeLa cells. Pretreatment of HeLa cells with 3-methyladenine (5 mmol/L) blocked the cytotoxic effect of fucoxanthin as well as fucoxanthin-induced autophagy.Conclusion:Fucoxanthin exerts autophagy-dependent cytotoxic effect in HeLa cells via inhibition of Akt/mTOR signaling pathway.

Hou LL; Gao C; Chen L; Hu GQ; Xie SQ

2013-08-01

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Targeting Pro-Apoptotic TRAIL Receptors Sensitizes HeLa Cervical Cancer Cells to Irradiation-Induced Apoptosis  

International Nuclear Information System (INIS)

Purpose: To investigate the potential of irradiation in combination with drugs targeting the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptor (DR)4 and DR5 and their mechanism of action in a cervical cancer cell line. Methods and Materials: Recombinant human TRAIL (rhTRAIL) and the agonistic antibodies against DR4 and DR5 were added to irradiated HeLa cells. The effect was evaluated with apoptosis and cytotoxicity assays and at the protein level. Membrane receptor expression was measured with flow cytometry. Small-interfering RNA against p53, DR4, and DR5 was used to investigate their function on the combined effect. Results: rhTRAIL and the agonistic DR4 and DR5 antibodies strongly enhanced 10-Gy-induced apoptosis. This extra effect was 22%, 23%, and 29% for rhTRAIL, DR4, and DR5, respectively. Irradiation increased p53 expression and increased the membrane expression of DR5 and DR4. p53 suppression, as well as small-interfering RNA against DR5, resulted in a significant downregulation of DR5 membrane expression but did not affect apoptosis induced by irradiation and rhTRAIL. After small-interfering RNA against DR4, rhTRAIL-induced apoptosis and the additive effect of irradiation on rhTRAIL-induced apoptosis were abrogated, implicating an important role for DR4 in apoptosis induced through irradiation in combination with rhTRAIL. Conclusion: Irradiation-induced apoptosis is strongly enhanced by targeting the pro-apoptotic TRAIL receptors DR4 or DR5. Irradiation results in a p53-dependent increase in DR5 membrane expression. The sensitizing effect of rhTRAIL on irradiation in the HeLa cell line is, however especially mediated through the DR4 receptor.

2008-10-01

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Inhibition of clathrin by pitstop 2 activates the spindle assembly checkpoint and induces cell death in dividing HeLa cancer cells.  

UK PubMed Central (United Kingdom)

BACKGROUND: During metaphase clathrin stabilises the mitotic spindle kinetochore (K)-fibres. Many anti-mitotic compounds target microtubule dynamics. Pitstop 2™ is the first small molecule inhibitor of clathrin terminal domain and inhibits clathrin-mediated endocytosis. We investigated its effects on a second function for clathrin in mitosis. RESULTS: Pitstop 2 did not impair clathrin recruitment to the spindle but disrupted its function once stationed there. Pitstop 2 trapped HeLa cells in metaphase through loss of mitotic spindle integrity and activation of the spindle assembly checkpoint, phenocopying clathrin depletion and aurora A kinase inhibition. CONCLUSIONS: Pitstop 2 is therefore a new tool for investigating clathrin spindle dynamics. Pitstop 2 reduced viability in dividing HeLa cells, without affecting dividing non-cancerous NIH3T3 cells, suggesting that clathrin is a possible novel anti-mitotic drug target.

Smith CM; Haucke V; McCluskey A; Robinson PJ; Chircop M

2013-01-01

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Plexin-B1 is a target of miR-214 in cervical cancer and promotes the growth and invasion of HeLa cells.  

Science.gov (United States)

Plexin-B1, the receptor for Sema4D, has been reported to trigger multiple and sometimes opposing cellular responses in various types of tumor cells. It has been implicated in the regulation of tumor-cell survival, proliferation, angiogenesis, invasion and metastasis. However, the plexin-B1 gene expression and its regulatory mechanism in cervical cancer remain unclear. The present study shows that plexin-B1 is over-expressed in cervical tumor tissues compared to normal cervical tissues by immunohistochemistry, Western blotting and quantitative RT-PCR. The expression of plexin-B1 is significantly associated with cervical tumor metastasis and invasion according to the analysis of the clinicopathologic data. Plexin-B1 also promotes proliferation, migration and invasion in human cervical cancer HeLa cells. We also found that the plexin-B1 levels are inversely correlated with miR-214 amounts in both cervical cancer tissues and HeLa cells. And miR-214 expression level is also associated with metastasis and invasion of cervical tumor. Furthermore, we demonstrate that plexin-B1 is inhibited by miR-214 through a miR-214 binding site within the 3'UTR of plexin-B1 in HeLa cells. Ectopic expression of miR-214 could inhibit the proliferation capacity, migration and invasion ability of HeLa cells. Our findings suggest that plexin-B1, a target of miR-214, may function as an oncogene in human cervical cancer HeLa cells. PMID:21216304

Qiang, Ran; Wang, Fang; Shi, Li-Ying; Liu, Min; Chen, Shuang; Wan, Hai-Ying; Li, Yi-Xuan; Li, Xin; Gao, Song-Yuan; Sun, Bao-Cun; Tang, Hua

2011-01-07

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DYTOGENETIC ANALYSIS OF HELA AND CHANG CELLS  

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Full Text Available Based on the evaluation of two human cell lines, Hela and Chang, abeuploidy and several marker chromosomes were found in both cells. The morphological characteristic of marker chromosomes of Chang cells was distinctly different from HeLa. Certain submetacentric marker chromosome was frequently present among 80% of marker chromosomes of Chang cells which distinguished this line from HeLa, which showed the various identifiable marker chromosomes. This evidence clearly established the different etiology of these two human cell lines.

N.Lzadian; H.Sussman

1982-01-01

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Apoptosis induction and G2/M arrest of 2-methyl-1,3,6-trihydroxy-9,10-anthraquinone from Rubia yunnanensis in human cervical cancer Hela cells.  

Science.gov (United States)

2-Methyl-1,3,6-trihydroxy-9,10-anthraquinone (MTA), one of the major components isolated from the traditional Chinese medicine Rubia yunnanensis, exhibited inhibitory activity on the proliferation of several human cancer cell lines. The results from an annexin V-FITC (fluoresein-5-isothiocyanate) apoptosis assay and DNA content analysis showed that MTA exerted cytotoxicity via apoptosis induction and G2/M cell cycle arrest in human cervical carcinoma HeLa cells. Further, MTA was found to induce apoptosis of HeLa cells through the mitochondria-mediated pathway. It caused the translocation of Bax to the mitochondria and release of cytochrome c into the cytosol, which caused the cleavage of caspase and poly(ADP-ribose) polymerase and finally triggered the apoptosis. Furthermore, the p53/p21/Cdc2-cyclin B1 signaling was found related to the G2/M arrest caused by MTA. The over-expression of p21 and down-expression of cyclin B1 caused by MTA inactivated the Cdc2-cyclin B1 complex of G2/M checkpoint and finally caused the G2/M arrest in HeLa cells. This study demonstrated that MTA is a potential anti-cancer component of R. yunnanensis, a folk anti-cancer herb used in Yunnan, China. PMID:23700797

Zeng, Guang-Zhi; Fan, Jun-Ting; Xu, Jun-Ju; Li, Yan; Tan, Ning-Hua

2013-04-01

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Apoptosis induction and G2/M arrest of 2-methyl-1,3,6-trihydroxy-9,10-anthraquinone from Rubia yunnanensis in human cervical cancer Hela cells.  

UK PubMed Central (United Kingdom)

2-Methyl-1,3,6-trihydroxy-9,10-anthraquinone (MTA), one of the major components isolated from the traditional Chinese medicine Rubia yunnanensis, exhibited inhibitory activity on the proliferation of several human cancer cell lines. The results from an annexin V-FITC (fluoresein-5-isothiocyanate) apoptosis assay and DNA content analysis showed that MTA exerted cytotoxicity via apoptosis induction and G2/M cell cycle arrest in human cervical carcinoma HeLa cells. Further, MTA was found to induce apoptosis of HeLa cells through the mitochondria-mediated pathway. It caused the translocation of Bax to the mitochondria and release of cytochrome c into the cytosol, which caused the cleavage of caspase and poly(ADP-ribose) polymerase and finally triggered the apoptosis. Furthermore, the p53/p21/Cdc2-cyclin B1 signaling was found related to the G2/M arrest caused by MTA. The over-expression of p21 and down-expression of cyclin B1 caused by MTA inactivated the Cdc2-cyclin B1 complex of G2/M checkpoint and finally caused the G2/M arrest in HeLa cells. This study demonstrated that MTA is a potential anti-cancer component of R. yunnanensis, a folk anti-cancer herb used in Yunnan, China.

Zeng GZ; Fan JT; Xu JJ; Li Y; Tan NH

2013-04-01

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Anticancer effects of the engineered stem cells transduced with therapeutic genes via a selective tumor tropism caused by vascular endothelial growth factor toward HeLa cervical cancer cells.  

UK PubMed Central (United Kingdom)

The aim of the present study was to investigate the therapeutic efficacy of genetically engineered stem cells (GESTECs) expressing bacterial cytosine deaminase (CD) and/ or human interferon-beta (IFN-?) gene against HeLa cervical cancer and the migration factors of the GESTECs toward the cancer cells. Anticancer effect of GESTECs was examined in a co-culture with HeLa cells using MTT assay to measure cell viability. A transwell migration assay was performed so as to assess the migration capability of the stem cells to cervical cancer cells. Next, several chemoattractant ligands and their receptors related to a selective migration of the stem cells toward HeLa cells were determined by real-time PCR. The cell viability of HeLa cells was decreased in response to 5-fluorocytosine (5-FC), a prodrug, indicating that 5-fluorouracil (5-FU), a toxic metabolite, was converted from 5-FC by CD gene and it caused the cell death in a co-culture system. When IFN-? was additionally expressed with CD gene by these GESTECs, the anticancer activity was significantly increased. In the migration assay, the GESTECs selectively migrated to HeLa cervical cancer cells. As results of real-time PCR, chemoattractant ligands such as MCP-1, SCF, and VEGF were expressed in HeLa cells, and several receptors such as uPAR, VEGFR2, and c-kit were produced by the GESTECs. These GESTECs transduced with CD gene and IFN-? may provide a potential of a novel gene therapy for anticervical cancer treatments via their selective tumor tropism derived from VEGF and VEGFR2 expressions between HeLa cells and the GESTECs.

Kim HS; Yi BR; Hwang KA; Kim SU; Choi KC

2013-09-01

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Isolation of Melittin from Iranian Honey Bee Venom and Investigation of Its Effect on Proliferation of Cervical Cancer- HeLa Cell Line  

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Full Text Available Introduction: Cervical cancer is the second prevalent cancer in developing countries and the sixth prevalent cancer in USA. Since conventional treatment methods are associated with detrimental side effects, searching for new drugs using natural ingredients is very important. Previous studies have shown that melittin (main component of honey bee venom) has anticancer properties along with the effect on cell membrane and activation of apoptosis. In this study, inhibitory effects of melittin on the viability and proliferation of cervical cancer cell line (HeLa) was investigated. Methods: Melittin was purified from honeybee venom using reversed-phase HPLC method. Then, biological activity of melittin was examined by hemolytic activity analysis on the red blood cells. In order to investigate whether melittin inhibits proliferation of HeLa cell, MTT assay was performed. HeLa cells were plated in a 96-well plate and treated with serially diluted concentrations of melittin for 12 and 24 hours. The viability of the cells was measured via MTT assay at 540nm. Results: Melittin showed a strong hemolytic activity (HD50=0.5 µg/ml) which can be reduced by FBS(HD50=2 µg/ml). Results of MTT assay indicated that melittin shows cytotoxic effect on cervical cancer cells with IC50 = 1.2 ug/ml at 12h incubation period. Conclusion: In this study, biological activity of melittin and inhibitory effect of FBS on hemolysis were determined via hemolytic activity analysis. MTT assay indicated that melittin induced cytotoxic effects in a dose dependent manner on cervical cancer cells and it also revealed dependence on incubation time as well.

H Zarinnahad; A Mahmoodzadeh; K Pooshang Bagheri; M Mahdavi; D Shahbazzadeh; A Moradi

2013-01-01

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The Cytotoxicity Mechanism of 6-Shogaol-Treated HeLa Human Cervical Cancer Cells Revealed by Label-Free Shotgun Proteomics and Bioinformatics Analysis.  

UK PubMed Central (United Kingdom)

Cervical cancer is one of the most common cancers among women in the world. 6-Shogaol is a natural compound isolated from the rhizome of ginger (Zingiber officinale). In this paper, we demonstrated that 6-shogaol induced apoptosis and G2/M phase arrest in human cervical cancer HeLa cells. Endoplasmic reticulum stress and mitochondrial pathway were involved in 6-shogaol-mediated apoptosis. Proteomic analysis based on label-free strategy by liquid chromatography chip quadrupole time-of-flight mass spectrometry was subsequently proposed to identify, in a non-target-biased manner, the molecular changes in cellular proteins in response to 6-shogaol treatment. A total of 287 proteins were differentially expressed in response to 24 h treatment with 15 ?M 6-shogaol in HeLa cells. Significantly changed proteins were subjected to functional pathway analysis by multiple analyzing software. Ingenuity pathway analysis (IPA) suggested that 14-3-3 signaling is a predominant canonical pathway involved in networks which may be significantly associated with the process of apoptosis and G2/M cell cycle arrest induced by 6-shogaol. In conclusion, this work developed an unbiased protein analysis strategy by shotgun proteomics and bioinformatics analysis. Data observed provide a comprehensive analysis of the 6-shogaol-treated HeLa cell proteome and reveal protein alterations that are associated with its anticancer mechanism.

Liu Q; Peng YB; Qi LW; Cheng XL; Xu XJ; Liu LL; Liu EH; Li P

2012-01-01

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Standardized flavonoid-rich fraction of Artemisia princeps Pampanini cv. Sajabal induces apoptosis via mitochondrial pathway in human cervical cancer HeLa cells.  

UK PubMed Central (United Kingdom)

ETHNOPHARMACOLOGICAL RELEVANCE: Artemisia princeps Pampanini is widely used in Eastern traditional medicine for the treatment of circulatory disorders, such as, dysmenorrhea, hematuria, hemorrhoids, and inflammation, and is also used to treat chronic conditions, such as, cancers, ulcers, and digestive disorders. AIM OF THE STUDY: The purpose of this study is to investigate the effect of a standardized flavonoid-rich fraction of Artemisia princeps Pampanini cv. Sajabal (FRAP) on the induction of apoptosis and the molecular mechanism involved in human cervical cancer HeLa cells. MATERIALS AND METHODS: Human cervical cancer HeLa cells were treated with FRAP and apoptosis was detected by cell morphologic observation, annexin-V-PI staning and western blot analysis on the expression of protein associated with cell death. RESULTS: FRAP led to the cleavages of caspase-3, -8, and -9 and the cleavage of poly (ADP-ribose) polymerase (PARP) in HeLa cells. Caspase-3 inhibitor (z-DEVD-fmk), caspase-8 inhibitor (z-IETD-fmk), caspase-9 inhibitor (z-LEHD), and broad caspase inhibitor (z-VAD-fmk) significantly suppressed the FRAP-induced accumulation of annexin V positive cells. Furthermore, it was found that FRAP caused a loss of mitochondrial membrane potential (MMP) and the release of cytochrome c to the cytosol. Furthermore, the overexpression of Bcl-xL significantly prevented FRAP-induced apoptosis, MMP changes, and the activations of caspase-3, -8, and -9. Interestingly, pretreatment with caspase-8 inhibitor significantly reduced the FRAP-induced activation of caspase-3 but not that of caspase-9, whereas the caspase-3 inhibitor, z-DEVD-fmk, markedly attenuated the FRAP-induced activation of caspase-8. In BALB/c(nu/nu) mice bearing a HeLa xenograft, FRAP dosed at 25 or 50mg/kg significantly inhibited tumor growth. CONCLUSION: Our results indicate caspase-mediated activation of the mitochondrial death pathway plays a critical role in the FRAP-induced apoptosis of HeLa cells and that FRAP inhibits the in vivo tumor growth of HeLa xenograft mice.

Ju HK; Lee HW; Chung KS; Choi JH; Cho JG; Baek NI; Chung HG; Lee KT

2012-05-01

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Hiding in plain view: Genetic profiling reveals decades old cross-contamination of bladder cancer cell line KU7 with HeLa.  

UK PubMed Central (United Kingdom)

PURPOSE: KU7 is a popular urothelial carcinoma cell line that was isolated from the bladder of a patient at the Keio University (KU) in 1980. It has subsequently been widely used in laboratories around the world. Here we describe how routine cell line authentication has revealed that KU7 was cross-contaminated almost 30 years ago with HeLa, a cervical carcinoma cell line. MATERIALS AND METHODS: Presumed KU7 clones dating from 1984 to 1999 were provided by MD Anderson Cancer Center (MDACC), the Vancouver Prostate Centre (VPC), Kyoto University, Tokyo Medical University and KU, and HeLa was purchased from the ATCC. Genomic DNA was isolated and short tandem repeat (STR) analysis was performed by the Characterized Cell Line Core Facility at MDACC, the Fragment Analysis Facility at Johns Hopkins University and the RIKEN Bioresource Center (Japan). Comparative genomic hybridization (CGH) was performed on the Agilent platform at the VPC. RESULTS: The STR profile of all KU7 clones was an exact match with HeLa. The CGH of all samples revealed an abundance of shared chromosomal aberrations. Slight differences in some genomic areas are explained by genomic drift occurring in different KU7 clones separated by many years. CONCLUSIONS: Our analysis identified that a cross-contamination of KU7 with HeLa occurred prior to 1984 at the source institution. All KU7 clones in the urologic literature should be considered HeLa and the experimental results should be viewed in this light. Our results emphasize the need to authenticate cell lines in oncologic research.

Jäger W; Horiguchi Y; Shah J; Hayashi T; Awrey S; Gust KM; Hadaschik BA; Matsui Y; Anderson S; Bell RH; Ettinger S; So AI; Gleave ME; Lee IL; Dinney CP; Tachibana M; McConkey DJ; Black PC

2013-03-01

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Tuning the cellular uptake and cytotoxicity properties of oligonucleotide intercalator-functionalized mesoporous silica nanoparticles with human cervical cancer cells HeLa.  

UK PubMed Central (United Kingdom)

A series of organically functionalized, MCM-41 type mesoporous silica nanoparticle materials (PAP-LP-MSN and AP-PAP-MSN) with different pore sizes (5.7 nm and 2.5 nm, respectively) were synthesized and characterized. We selectively decorated the exterior particle surface of PAP-LP-MSN and the interior pore surface of AP-PAP-MSN with an oligonucleotide intercalating phenanthridinium functionality. While phenanthridinium itself is a cell membrane impermeable molecule, we demonstrated that both phenanthridinium-immobilized PAP-LP-MSN and AP-PAP-MSN materials could indeed be internalized by live human cervical cancer cells (HeLa). We discovered that the PAP-LP-MSN nanoparticles with the phenanthridium groups located on the exterior surface were able to bind to cytoplasmic oligonucleotides, such as messenger RNAs, of HeLa cells resulting in severe cell growth inhibition. In contrast, the cytotoxicity of AP-PAP-MSN, where the same oligonucleotide intercalating molecules were anchored inside the pores, was significantly lowered upon the endocytosis by HeLa cells. We envision that this approach of combining the selective functionalization of the two different surfaces (exterior particle and interior pore surfaces) with morphology control of mesoporous silica nanoparticles would lead to a new generation of nanodevices with tunable biocompatibility and cell membrane trafficking properties for many biomedical applications.

Vivero-Escoto JL; Slowing II; Lin VS

2010-02-01

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Tuning the cellular uptake and cytotoxicity properties of oligonucleotide intercalator-functionalized mesoporous silica nanoparticles with human cervical cancer cells HeLa.  

Science.gov (United States)

A series of organically functionalized, MCM-41 type mesoporous silica nanoparticle materials (PAP-LP-MSN and AP-PAP-MSN) with different pore sizes (5.7 nm and 2.5 nm, respectively) were synthesized and characterized. We selectively decorated the exterior particle surface of PAP-LP-MSN and the interior pore surface of AP-PAP-MSN with an oligonucleotide intercalating phenanthridinium functionality. While phenanthridinium itself is a cell membrane impermeable molecule, we demonstrated that both phenanthridinium-immobilized PAP-LP-MSN and AP-PAP-MSN materials could indeed be internalized by live human cervical cancer cells (HeLa). We discovered that the PAP-LP-MSN nanoparticles with the phenanthridium groups located on the exterior surface were able to bind to cytoplasmic oligonucleotides, such as messenger RNAs, of HeLa cells resulting in severe cell growth inhibition. In contrast, the cytotoxicity of AP-PAP-MSN, where the same oligonucleotide intercalating molecules were anchored inside the pores, was significantly lowered upon the endocytosis by HeLa cells. We envision that this approach of combining the selective functionalization of the two different surfaces (exterior particle and interior pore surfaces) with morphology control of mesoporous silica nanoparticles would lead to a new generation of nanodevices with tunable biocompatibility and cell membrane trafficking properties for many biomedical applications. PMID:19932923

Vivero-Escoto, Juan L; Slowing, Igor I; Lin, Victor S-Y

2009-11-24

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The Cytotoxic and Synergistic Effects of Flavonoid Derivatives on Doxorubicin Cytotoxicity in Hela, MDA-MB-231, and HT-29 Cancer Cells  

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Full Text Available Background: Flavonoids have a variety of biological activities, such as anti-allergic,anti-inflammatory, anti-oxidative, free radical scavenging, and anti-mutagenic.Methods: The cytotoxic effects of three synthesized flavonoid derivatives (K3, K4and K5) were evaluated against Hela , MDA-MB -231 and HT-29 cancer cells usingMTT assay.Results: The results showed that these flavonoids were not cytotoxic at any testedconcentrations (0.1, 0.05, 0.01, and 0.001 mM). To evaluate the possible synergisticeffect of synthetic flavonoids with chemotherapeutic agent, the compound (K4) wasexamined against Hela and MDA-MB-231 cells in combination with differentconcentrations of doxorubicin (0.01 and 0.001mM). This combination treatmentsignificantly increased the cytotoxicity of doxorubicin (P < 0.05).Conclusion: Synthesized flavonoids could be used either in combination therapywith other chemotherapeutic agents or used as antioxidants in food supplements.

Hojjat Sadeghi-Aliabadi; Hossein Mosavi; Mina Mirian; Saeed Kakhki; Afshin Zarghi

2012-01-01

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Anthocyanins from Vitis coignetiae Pulliat Inhibit Cancer Invasion and Epithelial-Mesenchymal Transition, but These Effects Can Be Attenuated by Tumor Necrosis Factor in Human Uterine Cervical Cancer HeLa Cells.  

UK PubMed Central (United Kingdom)

Recently we have demonstrated that anthocyanins from fruits of Vitis coignetiae Pulliat (AIMs) have anticancer effects. Here, we investigate the effects of AIMs on cell proliferation and invasion as well as epithelial-mesenchymal transition (EMT) which have been linked to cancer metastasis in human uterine cervical cancer HeLa cells. AIMs inhibited the invasion of HeLa cells in a dose-dependent manner. AIMs inhibited MMP-9 expression in a dose-dependent manner. AIMs inhibited the motility of HeLa cells in a wound healing test. AIMs still suppressed NF- ? B activation induced by TNF. AIMs also inhibited EMT in HeLa cells. AIMs suppressed vimentin, N-cadherin, and ?-catenin expression and induced E-cadherin. AIMs also suppressed expression of ?-catenin and Snail, which was regulated by GSK-3. These effects of AIMs were also limited in the HeLa cells treated with TNF. In conclusion, this study indicates that AIMs have anticancer effects by suppressing NF- ? B-regulated genes and EMT, which relates to suppression of I ? B ? phosphorylation and GSK-3 activity, respectively. However, the effects of AIMs were attenuated in the TNF-high condition.

Lu JN; Lee WS; Yun JW; Kim MJ; Kim HJ; Kim DC; Jeong JH; Choi YH; Kim GS; Ryu CH; Shin SC

2013-01-01

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Role of altholactone in inducing type II apoptosis signalling pathway and expression of cancer-related genes in cervical carcinoma HeLa cell line.  

UK PubMed Central (United Kingdom)

Goniothalamus species (Annonaceae) is a shrub that grows in the rainforest of tropical Asia. Several compounds have been isolated and exhibit the potential use for cancer treatment. In this work, altholactone isolated from Goniothalamus macrophyllus was investigated for its cytotoxicity, apoptosis signalling and the expression of cancer-related genes in the cervical carcinoma HeLa cells. Cytotoxicity was evaluated by MTT assay. Apoptotic characteristics were evaluated by morphological studies. Caspase-3 activity was detected using a fluorogenic substrate. Cytochrome c release from mitochondria and protein Bid were determined by Western blotting and cancer-related genes expression by RT-PCR. The results demonstrated that altholactone was cytotoxic to HeLa (IC50 ?=?9.6??g/mL), and apoptotic cell death was manifested by appearance of chromatin condensation and caspase-3 activation, which was inhibited by specific inhibitors of both caspase-8 and -9. Release into the cytosol of cytochrome and cleavage of Bid occurred. Altholactone also caused a decrease in bcl-2 and an increase in p53 expression. These unique properties of altholactone suggest a potential for cancer chemotherapy.

Uthaisang-Tanechpongtamb W; Sriyabhaya P; Wilairat P

2013-05-01

 
 
 
 
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The Potentized Homeopathic Drug, Lycopodium clavatum (5C and 15C) Has Anti-cancer Effect on HeLa Cells In Vitro.  

Science.gov (United States)

Cancer is a disease that needs a multi-faceted approach from different systems of medicine. The purpose of this study was to evaluate whether homeopathically-potentized ultra-high dilutions of Lycopodium Clavatum (LC-5C and LC-15C, respectively) have any anti-cancer effects on HeLa cells. Cells were exposed to either LC-5C (diluted below Avogadro's limit, i.e., 10(-10)) or LC-15C (diluted beyond Avogadro's limit, i.e., 10(-30)) (drug-treated) or to 30% succussed ethanol ("vehicle" of the drug). The drug-induced modulation in the percent cell viability, the onset of apoptosis, and changes in the expressions of Bax, Bcl2, caspase 3, and Apaf proteins in inter-nucleosomal DNA, in mitochondrial membrane potentials and in the release of cytochrome-c were analyzed by utilizing different experimental protocols. Results revealed that administration of LC-5C and LC-15C had little or no cytotoxic effect in normal peripheral blood mononuclear cells, but caused considerable cell death through apoptosis in cancer (HeLa) cells, which was evident from the induction of DNA fragmentation, the increases in the expressions of protein and mRNA of caspase 3 and Bax, and the decreases in the expressions of Bcl2 and Apaf and in the release of cytochrome-c. Thus, the highly-diluted, dynamized homeopathic remedies LC-5C and LC-15C demonstrated their capabilities to induce apoptosis in cancer cells, signifying their possible use as supportive medicines in cancer therapy. PMID:23972240

Samadder, Asmita; Das, Sreemanti; Das, Jayeeta; Paul, Avijit; Boujedaini, Naoual; Khuda-Bukhsh, Anisur Rahman

2013-04-28

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The Potentized Homeopathic Drug, Lycopodium clavatum (5C and 15C) Has Anti-cancer Effect on HeLa Cells In Vitro.  

UK PubMed Central (United Kingdom)

Cancer is a disease that needs a multi-faceted approach from different systems of medicine. The purpose of this study was to evaluate whether homeopathically-potentized ultra-high dilutions of Lycopodium Clavatum (LC-5C and LC-15C, respectively) have any anti-cancer effects on HeLa cells. Cells were exposed to either LC-5C (diluted below Avogadro's limit, i.e., 10(-10)) or LC-15C (diluted beyond Avogadro's limit, i.e., 10(-30)) (drug-treated) or to 30% succussed ethanol ("vehicle" of the drug). The drug-induced modulation in the percent cell viability, the onset of apoptosis, and changes in the expressions of Bax, Bcl2, caspase 3, and Apaf proteins in inter-nucleosomal DNA, in mitochondrial membrane potentials and in the release of cytochrome-c were analyzed by utilizing different experimental protocols. Results revealed that administration of LC-5C and LC-15C had little or no cytotoxic effect in normal peripheral blood mononuclear cells, but caused considerable cell death through apoptosis in cancer (HeLa) cells, which was evident from the induction of DNA fragmentation, the increases in the expressions of protein and mRNA of caspase 3 and Bax, and the decreases in the expressions of Bcl2 and Apaf and in the release of cytochrome-c. Thus, the highly-diluted, dynamized homeopathic remedies LC-5C and LC-15C demonstrated their capabilities to induce apoptosis in cancer cells, signifying their possible use as supportive medicines in cancer therapy.

Samadder A; Das S; Das J; Paul A; Boujedaini N; Khuda-Bukhsh AR

2013-08-01

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Apoptosis-inducing and Antitumor Activity of Neolignans Isolated from Magnolia officinalis in HeLa Cancer Cells.  

UK PubMed Central (United Kingdom)

Two neolignans, 4'-methoxymagndialdehyde (1) and magnaldehyde B (2), were isolated from the stem bark of Magnolia officinalis (Magnoliaceae), evaluated for apoptosis-inducing effects in human cervical epitheloid carcinoma HeLa cells. The apoptosis-inducing activity of compounds 1 and 2 were assessed by DNA content using flow cytometric analysis. In the immunoblotting analysis, the treatment with 1 and 2 resulted in the cleavage of procaspase-8 and -3 and poly(ADP-ribose)polymerase into active forms. In addition, in vivo, the administration of 2 to Lewis lung carcinoma-inoculated mice evidenced a significant inhibition of tumor growth (volume) with reduction of 28.7% at concentration of 20 mg/kg, as compared with the control mice. These findings suggest that 2 can inhibit the proliferation of tumor cells, and might be an anti-tumoric agent. Copyright © 2012 John Wiley & Sons, Ltd.

Youn UJ; Fatima N; Chen QC; Chae S; Hung TM; Min BS

2013-09-01

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Inhibin-betaA and -betaB subunits in normal and malignant glandular epithelium of uterine cervix and HeLa cervical cancer cell line.  

UK PubMed Central (United Kingdom)

INTRODUCTION: Inhibins, dimeric peptide hormones composed of an alpha-subunit and one of two possible beta-subunits (betaA or betaB), exhibit substantial roles in human reproduction and in endocrine-responsive tumors. However, it is still unclear if normal and cancerous cervical glandular epithelial cells as well as cervical cancer cell lines of glandular origin express the inhibin-betaA and -betaB subunits. MATERIALS AND METHODS: Normal cervical tissue samples and a total of 10 specimens of well-differentiated adenocarcinomas of the human cervix were analyzed for inhibin-betaA and -betaB subunit expression by immunohistochemical analysis. Additionally, the cervical carcinoma cell line HeLa was analyzed by immunofluorescence and RT-PCR analysis for the expression of inhibin subunits. RESULTS: Immunolabeling of normal and malignant glandular epithelium of human cervical tissue revealed a positive staining reaction for the inhibin-betaA and -betaB subunits. Additionally, the cancer cell line HeLa synthesized both inhibin subunits. When compared to the normal cervical glandular epithelium, the expression of the inhibin beta subunits became significantly reduced in cervical adenocarcinoma tissues. DISCUSSION: In conclusion, we demonstrated a strong, though differential expression pattern of inhibin-betaA and -betaB subunits in normal and malignant glandular epithelial cells of the human uterine cervix. Although the physiological role of inhibins is still quite unclear in cervical tissue, the expression of inhibin-beta-subunits might play an important role in cervical cancer carcinogenesis, since they are significantly down-regulated during pathogenesis in cervical adenocarcinomas.

Burges A; Shabani N; Brüning A; Mylonas I

2011-10-01

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Effects of artesunate combining with radiation on apoptosis in nude mice transplanted with HeLa cells of cervical cancer  

International Nuclear Information System (INIS)

Objective: To investigate the effect of Artesunate combining with radiation on apoptosis in transplanted tumors. Methods: HeLa cells were inoculated into the nude mice to develop a tumor model. Mice were randomized into four groups as the control group, the Artesunate group,the irradiation group and the combination group when average volume of tumor achieved about 5 mm x 5 mm x 5 mm. During the period of treatment, the volume of tumors was measured per 2 days. After 14 days treatment, the mice were killed and tumor tissues were harvest, the tumor size and weight were measured, tumor inhibitory rate calculated and TUNEL assay was used to analysis the apoptosis of tumor tissue. Results: The tumor weight in combination group was significantly lower than that than in the irradiation group [(0.64 ± 0.11) gvs (1.31 ± 0.58) g] (P

2011-01-01

46

The Aqueous Extract of Ficus religiosa Induces Cell Cycle Arrest in Human Cervical Cancer Cell Lines SiHa (HPV-16 Positive) and Apoptosis in HeLa (HPV-18 Positive)  

Science.gov (United States)

Natural products are being extensively explored for their potential to prevent as well as treat cancer due to their ability to target multiple molecular pathways. Ficus religiosa has been shown to exert diverse biological activities including apoptosis in breast cancer cell lines. In the present study, we report the anti-neoplastic potential of aqueous extract of F. religiosa (FRaq) bark in human cervical cancer cell lines, SiHa and HeLa. FRaq altered the growth kinetics of SiHa (HPV-16 positive) and HeLa (HPV-18 positive) cells in a dose-dependent manner. It blocked the cell cycle progression at G1/S phase in SiHa that was characterized by an increase in the expression of p53, p21 and pRb proteins with a simultaneous decrease in the expression of phospho Rb (ppRb) protein. On the other hand, in HeLa, FRaq induced apoptosis through an increase in intracellular Ca2+ leading to loss of mitochondrial membrane potential, release of cytochrome-c and increase in the expression of caspase-3. Moreover, FRaq reduced the migration as well as invasion capability of both the cervical cancer cell lines accompanied with downregulation of MMP-2 and Her-2 expression. Interestingly, FRaq reduced the expression of viral oncoproteins E6 and E7 in both the cervical cancer cell lines. All these data suggest that F. religiosa could be explored for its chemopreventive potential in cervical cancer.

Choudhari, Amit S.; Suryavanshi, Snehal A.; Kaul-Ghanekar, Ruchika

2013-01-01

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The Aqueous Extract of Ficus religiosa Induces Cell Cycle Arrest in Human Cervical Cancer Cell Lines SiHa (HPV-16 Positive) and Apoptosis in HeLa (HPV-18 Positive).  

UK PubMed Central (United Kingdom)

Natural products are being extensively explored for their potential to prevent as well as treat cancer due to their ability to target multiple molecular pathways. Ficus religiosa has been shown to exert diverse biological activities including apoptosis in breast cancer cell lines. In the present study, we report the anti-neoplastic potential of aqueous extract of F. religiosa (FRaq) bark in human cervical cancer cell lines, SiHa and HeLa. FRaq altered the growth kinetics of SiHa (HPV-16 positive) and HeLa (HPV-18 positive) cells in a dose-dependent manner. It blocked the cell cycle progression at G1/S phase in SiHa that was characterized by an increase in the expression of p53, p21 and pRb proteins with a simultaneous decrease in the expression of phospho Rb (ppRb) protein. On the other hand, in HeLa, FRaq induced apoptosis through an increase in intracellular Ca(2+) leading to loss of mitochondrial membrane potential, release of cytochrome-c and increase in the expression of caspase-3. Moreover, FRaq reduced the migration as well as invasion capability of both the cervical cancer cell lines accompanied with downregulation of MMP-2 and Her-2 expression. Interestingly, FRaq reduced the expression of viral oncoproteins E6 and E7 in both the cervical cancer cell lines. All these data suggest that F. religiosa could be explored for its chemopreventive potential in cervical cancer.

Choudhari AS; Suryavanshi SA; Kaul-Ghanekar R

2013-01-01

48

The Aqueous Extract of Ficus religiosa Induces Cell Cycle Arrest in Human Cervical Cancer Cell Lines SiHa (HPV-16 Positive) and Apoptosis in HeLa (HPV-18 Positive).  

Science.gov (United States)

Natural products are being extensively explored for their potential to prevent as well as treat cancer due to their ability to target multiple molecular pathways. Ficus religiosa has been shown to exert diverse biological activities including apoptosis in breast cancer cell lines. In the present study, we report the anti-neoplastic potential of aqueous extract of F. religiosa (FRaq) bark in human cervical cancer cell lines, SiHa and HeLa. FRaq altered the growth kinetics of SiHa (HPV-16 positive) and HeLa (HPV-18 positive) cells in a dose-dependent manner. It blocked the cell cycle progression at G1/S phase in SiHa that was characterized by an increase in the expression of p53, p21 and pRb proteins with a simultaneous decrease in the expression of phospho Rb (ppRb) protein. On the other hand, in HeLa, FRaq induced apoptosis through an increase in intracellular Ca(2+) leading to loss of mitochondrial membrane potential, release of cytochrome-c and increase in the expression of caspase-3. Moreover, FRaq reduced the migration as well as invasion capability of both the cervical cancer cell lines accompanied with downregulation of MMP-2 and Her-2 expression. Interestingly, FRaq reduced the expression of viral oncoproteins E6 and E7 in both the cervical cancer cell lines. All these data suggest that F. religiosa could be explored for its chemopreventive potential in cervical cancer. PMID:23922932

Choudhari, Amit S; Suryavanshi, Snehal A; Kaul-Ghanekar, Ruchika

2013-07-26

49

The effects of ionizing radiation combined with autophagy inducers or inhibitors or inhibitors on human cervical cancer hela cells  

International Nuclear Information System (INIS)

Objective: To detect the effects of ionizing radiation combined with autophagy inhibitors and inducers on the proliferation of human cervical cancer cell line. Methods: MTT and flowcytometry (FCM) were used to detect the surviving and proliferation of human cervical cancer cells,and analysis of the relationship of dose-effect and time-effect was made. Results: With the increase of irradiation doses (2, 4, 6, 8 and 10 Gy) and the elongation of irradiation time (24, 48 and 72 h), the inhibiting effect of ionizing radiation on the proliferation of human cervical cancer cells increased (P

2010-01-01

50

Irradiation And Papillomavirus E2 Proteins On Hela Cells  

International Nuclear Information System (INIS)

[en] Exposure to relatively high doses ionizing radiation activates cellular responses that impair cell survival. These responses, for which the p53 protein plays a central role, form the basis for cancer radiotherapy. However, the efficacy of radiation treatments on cell killing is often reduced as a consequence of the frequent inactivation of the p53 protein in cancer cells. Loss of p53 protein is associated with later stages of most human tumors and resistance to anticancer agents. Carcinomas are frequent malignant tumors in humans. The majority of cervical carcinomas are etiologically linked to the presence of HPV virus (Human Papillomavirus). In carcinoma tumor cells, as well as in their derived-cell lines such as HeLa cells, the p53 protein is generally not detected due to its degradation by the product of the HPV-associated oncogenic E6 gene. Another characteristic of HPV-positive cervical cancer cells is the loss of the regulatory viral E2 gene expression as a consequence of viral DNA integration into the cellular genome. Reintroduction of E2 expression in HeLa cells reactivates p53, due to a negative effect on the expression of E6 protein, with a concomitant arrest of cell proliferation at the phase G1 of the cell cycle and delay in cell division via the repression of E2F-target genes. To elucidate whether reactivation of p53 would improve the cell killing effect of ionizing radiation in cancer cells, we studied the combined effects of radiation and E2 expression on the cell cycle distribution in HeLa cells

2005-01-01

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Breast Cancer Resistance Protein-Mediated Efflux of Luteolin Glucuronides in HeLa Cells Overexpressing UDP-Glucuronosyltransferase 1A9.  

UK PubMed Central (United Kingdom)

PURPOSE: UDP-glucuronosyltransferases (UGTs) are responsible for the formation of glucuronides of polyphenolic flavonoids. This study investigated the UGT1A9-mediated glucuronidation of luteolin and the kinetics of luteolin glucuronide efflux. METHOD: HeLa cells overexpressing UGT1A9 (HeLa-UGT1A9) were used to determine the kinetics of breast cancer resistance protein (BCRP)-mediated transport of luteolin glucuronides. Human UGT isoforms were used to determine glucuronidation rates. RESULTS: UGT1A9 was found to catalyze the production of four luteolin glucuronides, including three known monoglucuronides and a novel 3', 4'-diglucuronide. Ko143, a potent specific inhibitor of BCRP, significantly inhibited efflux of luteolin monoglucuronides from HeLa1A9 cells and increased their intracellular levels in a dose-dependent manner. The formation of luteolin diglucuronide was observed when intracellular concentration of total monoglucuronides went above 0.07 nM. CONCLUSIONS: Intracellular accumulation of diglucuronide was detected at high monoglucuronide concentrations (>0.07 nM). Diglucuronide production is speculated to be a compensatory pathway for luteolin disposition.

Tang L; Li Y; Chen WY; Zeng S; Dong LN; Peng XJ; Jiang W; Hu M; Liu ZQ

2013-10-01

52

Comparative proteomic analysis Of indioside d-triggered cell death in HeLa cells  

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Medicinal plants represent a rich source of cancer drug leads. Indioside D, a furostanol glycoside isolated from Solanum mammosum, was found to possess antiproliferative activity toward a panel of human cancer cell lines. Proteomic analysis of indioside D-treated HeLa cells revealed profound protein...

Wong, CC; Wang, Y; Cheng, KW; Chiu, JF; He, QY; Chen, F

53

S39, a novel Aurora B kinase inhibitor, shows potent antineoplastic activity in human Hela cervical cancer cell line.  

UK PubMed Central (United Kingdom)

Aurora kinases, frequently detected to be over-expressing in human tumors, regulate many essential events during mitosis progression and have been regarded as potentially important targets for cancer therapy. S39 is a novel potent inhibitor of Aurora B kinase with the IC50 90.07 nM in the biochemical assay in an ATP competitive manner. S39 treatment on human tumor cells can inhibit the phosphorylation of Histone H3 (Ser10), a direct downstream substrate of Aurora B kinase, indicating S39 inhibits endogenous Aurora B kinase activity in cell-based level. Furthermore, S39 treatment blocks cell proliferation, inhibits colony formation and induces apoptosis in a wide range of human tumor cell lines. These results indicate that S39 is a potential lead compound to be an Aurora B inhibitor.

Li J; Lang Q; Zhang H; Huang Q; Yu L

2013-06-01

54

Effects of different concentrations of juglone on the proliferation and apoptosis of HeLa cells  

Directory of Open Access Journals (Sweden)

Full Text Available Objective ?To study the effect of different concentrations of juglone on the proliferation and apoptosis of human cervical cancer cells (HeLa), and explore the anti-tumor effect of juglone in vitro. Methods ?HeLa cells were cultured in vitrofor 24 h and then divided into control group and juglone-treated group. The cells in juglone-treated group were cultured again with 10, 20, 50, 100 and 200?mol/L juglone, respectively, for 24h. The morphological changes in HeLa cells were observed with inverted microscope. The proliferation of HeLa cells was assessed detected by methyl thiazolyl tetrazolium (MTT) assay. The cell cycle and apoptosis were observed by flow cytometry (FCM). The expression of caspase-3 in HeLa cells was determined by caspase-3 colorimetric assay kit. Results ?Under morphological observation, it was found that different concentrations of juglone led to change in cell shape. MTT assay showed that juglone significantly inhibited the growth of HeLa cells in a dose-dependent manner (P<0.05 or P<0.01). FCM assay indicated that the percentage of the cell cycles arresting at G2/M phase after treatment with above concentrations of juglone for 24 h were increased to 12.92%, 16.23%, 23.64%, 34.22% and 52.64% from 7.5%, respectively (P<0.05 or P<0.01). The caspase-3 activity in juglone-treated HeLa cells remarkably increased in a dose-dependent manner as compared with control group. Conclusion ?Various concentrations of juglone can inhibit the proliferation and induce the apoptosis of HeLa cells in a dose-dependent manner, implying that juglone may have anti-tumor effect.

Wei ZHANG; Xing-yu ZHAO; Yan LI; Jun-jie XU; Yan-xia JIANG; Shi-jie LV

2013-01-01

55

[Effect of methotrexate (MTX) on neoplastic (HeLa line) and normal (lymphocyte) cells  

UK PubMed Central (United Kingdom)

An equipment simulating the pharmacodynamics of anti-cancer drugs under time lapse microcinematography is described. It consists of an inverted microscope furnished with an incubating chamber, a motion-picture camera, an intervalometer, a gradient forming device and peristaltic pump. In this work, the equipment is used to study the effect of methotrexate (MTX) on cultures of HeLa cells and HeLa cell-sensitized lymphoblasts. MTX inhibits cell reproduction of both, HeLa cells and lymphoblasts, prolongs twice the generation time of the cancer cells, and produces a progressive cell degeneration. MTX produces also an early degeneration of lymphoblasts. All these data are studied by individual frame analysis of the films.

Ruano A; Alvarez-Rodríguez Y; Alvarez-Noves J; Valladares Y

1982-01-01

56

Daphnoretin-induced apoptosis in HeLa cells: a possible mitochondria-dependent pathway.  

UK PubMed Central (United Kingdom)

Daphnoretin is a bicoumarin compound isolated from a natural product, Wikstroemia indica, which has been used to treat many diseases. It has strong antiviral and anti-tumor activities. Taking the anti-tumor activity of daphnoretin as a starting point, the present study aimed to test the pro-apoptotic effect of daphnoretin and its underlying mechanism in HeLa cells. The inhibitory effects of daphnoretin on viability and proliferation of HeLa cells were determined by the MTT assay. Daphnoretin-induced apoptotic morphological changes were analyzed by mitochondrial membrane potential and Hoechst staining. The number and stage of apoptotic HeLa cells were determined by flow cytometry. Gene expression was determined by reverse-transcription polymerase chain reaction. Protein expression was determined by western blot. The caspase activity of HeLa cells was detected by a caspase-3 and caspase-9 colorimetric assay kit. We found that daphnoretin significantly inhibited HeLa cells' viability by the MTT assay and flow cytometry. The nuclei of the apoptotic cells exhibited strong, blue fluorescence in Hoechst staining. Bax mRNA and protein levels were increased while bcl-2 mRNA levels were decreased after daphnoretin treatment. Daphnoretin also activated both caspase-3 and caspase-9. These findings suggest that daphnoretin promotes apoptosis of HeLa cells in a mitochondria-mediated way. Daphnoretin therefore has potential to be a promising drug to treat uterine cervix cancer.

Yang ZY; Kan JT; Cheng ZY; Wang XL; Zhu YZ; Guo W

2013-10-01

57

Pardaxin-induced apoptosis enhances antitumor activity in HeLa cells.  

UK PubMed Central (United Kingdom)

Pardaxin, a pore-forming antimicrobial peptide that encodes 33 amino acids was isolated from the Red Sea Moses sole, Pardachirus mamoratus. In this study, we investigated its antitumor activity in human fibrosarcoma (HT-1080) cells and epithelial carcinoma (HeLa) cells. In vitro results showed that the synthetic pardaxin peptide had antitumor activity in these two types of cancer cells and that 15?g/ml pardaxin did not lyse human red blood cells. Moreover, this synthetic pardaxin inhibited the proliferation of HT1080 cells in a dose-dependent manner and induced programmed cell death in HeLa cells. DNA fragmentation and increases in the subG1 phase and caspase 8 activities suggest that pardaxin caused HeLa cell death by inducing apoptosis, but had a different mechanism in HT1080 cells.

Hsu JC; Lin LC; Tzen JT; Chen JY

2011-06-01

58

Comparative proteomic analysis of indioside D-triggered cell death in HeLa cells.  

Science.gov (United States)

Medicinal plants represent a rich source of cancer drug leads. Indioside D, a furostanol glycoside isolated from Solanum mammosum, was found to possess antiproliferative activity toward a panel of human cancer cell lines. Proteomic analysis of indioside D-treated HeLa cells revealed profound protein changes related to energy production and oxidative stress, suggesting that mitochondria dysfunction plays a role in indioside D-induced apoptosis. Indioside D caused a rapid dissipation of mitochondrial transmembrane potential (DeltaPsim) and the generation of reactive oxygen species (ROS), leading to the activation of caspase-dependent apoptotic cell death. The Fas death receptor pathway was also activated following indioside D treatment, and triggered the activation of caspase-8 and cleavage of Bid, which also acted through the mitochondrial apoptosis pathway. These results suggest that indioside D induced apoptosis in HeLa cells via both intrinsic and extrinsic cell death pathways. PMID:18376857

Wong, Chi Chun; Wang, Ying; Cheng, Ka-Wing; Chiu, Jen-Fu; He, Qing-Yu; Chen, Feng

2008-04-01

59

Dietary Flavonoids Sensitize HeLa Cells to Tumor Necrosis Factor-Related Apoptosis-Inducing Ligand (TRAIL)  

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Full Text Available TRAIL is a promising candidate for cancer therapeutics that preferentially induces apoptosis in cancer cells. The combined treatment flavonoids with TRAIL might be promising as a chemoprevention and/or new therapy against malignant tumors. We examined the cytotoxic effect of dietary flavonoids in combination with TRAIL on HeLa cells. It was found that treatment with noncytotoxic concentration of some flavonoids significantly sensititizes to TRAIL induced death in HeLa cells. Our study demonstrated that flavone, apigenin and genistein markedly augmented TRAIL mediated cytotoxicity against HeLa, whereas kaempferol and quercetin produced no effect.

Ewelina Szliszka; Zenon P. Czuba; Katarzyna Jernas; Wojciech Król

2008-01-01

60

Phorbol esters from Jatropha meal triggered apoptosis, activated PKC-?, caspase-3 proteins and down-regulated the proto-oncogenes in MCF-7 and HeLa cancer cell lines.  

UK PubMed Central (United Kingdom)

Jatropha meal produced from the kernel of Jatropha curcas Linn. grown in Malaysia contains phorbol esters (PEs). The potential benefits of PEs present in the meal as anticancer agent are still not well understood. Hence, this study was conducted to evaluate the cytotoxic effects and mode of actions of PEs isolated from Jatropha meal against breast (MCF-7) and cervical (HeLa) cancer cell lines. Isolated PEs inhibited cells proliferation in a dose-dependent manner of both MCF-7 and HeLa cell lines with the IC?? of 128.6 ± 2.51 and 133.0 ± 1.96 µg PMA equivalents/mL respectively, while the values for the phorbol 12-myristate 13-acetate (PMA) as positive control were 114.7 ± 1.73 and 119.6 ± 3.73 µg/mL, respectively. Microscopic examination showed significant morphological changes that resemble apoptosis in both cell lines when treated with PEs and PMA at IC?? concentration after 24 h. Flow cytometry analysis and DNA fragmentation results confirmed the apoptosis induction of PEs and PMA in both cell lines. The PEs isolated from Jatropha meal activated the PKC-? and down-regulated the proto-oncogenes (c-Myc, c-Fos and c-Jun). These changes probably led to the activation of Caspase-3 protein and apoptosis cell death occurred in MCF-7 and HeLa cell lines upon 24 h treatment with PEs and PMA. Phorbol esters of Jatropha meal were found to be promising as an alternative to replace the chemotherapeutic drugs for cancer therapy.

Oskoueian E; Abdullah N; Ahmad S

2012-01-01

 
 
 
 
61

Phorbol esters from Jatropha meal triggered apoptosis, activated PKC-?, caspase-3 proteins and down-regulated the proto-oncogenes in MCF-7 and HeLa cancer cell lines.  

Science.gov (United States)

Jatropha meal produced from the kernel of Jatropha curcas Linn. grown in Malaysia contains phorbol esters (PEs). The potential benefits of PEs present in the meal as anticancer agent are still not well understood. Hence, this study was conducted to evaluate the cytotoxic effects and mode of actions of PEs isolated from Jatropha meal against breast (MCF-7) and cervical (HeLa) cancer cell lines. Isolated PEs inhibited cells proliferation in a dose-dependent manner of both MCF-7 and HeLa cell lines with the IC?? of 128.6 ± 2.51 and 133.0 ± 1.96 µg PMA equivalents/mL respectively, while the values for the phorbol 12-myristate 13-acetate (PMA) as positive control were 114.7 ± 1.73 and 119.6 ± 3.73 µg/mL, respectively. Microscopic examination showed significant morphological changes that resemble apoptosis in both cell lines when treated with PEs and PMA at IC?? concentration after 24 h. Flow cytometry analysis and DNA fragmentation results confirmed the apoptosis induction of PEs and PMA in both cell lines. The PEs isolated from Jatropha meal activated the PKC-? and down-regulated the proto-oncogenes (c-Myc, c-Fos and c-Jun). These changes probably led to the activation of Caspase-3 protein and apoptosis cell death occurred in MCF-7 and HeLa cell lines upon 24 h treatment with PEs and PMA. Phorbol esters of Jatropha meal were found to be promising as an alternative to replace the chemotherapeutic drugs for cancer therapy. PMID:22964499

Oskoueian, Ehsan; Abdullah, Norhani; Ahmad, Syahida

2012-09-10

62

Phorbol Esters from Jatropha Meal Triggered Apoptosis, Activated PKC-?, Caspase-3 Proteins and Down-Regulated the Proto-Oncogenes in MCF-7 and HeLa Cancer Cell Lines  

Directory of Open Access Journals (Sweden)

Full Text Available Jatropha meal produced from the kernel of Jatropha curcas Linn. grown in Malaysia contains phorbol esters (PEs). The potential benefits of PEs present in the meal as anticancer agent are still not well understood. Hence, this study was conducted to evaluate the cytotoxic effects and mode of actions of PEs isolated from Jatropha meal against breast (MCF-7) and cervical (HeLa) cancer cell lines. Isolated PEs inhibited cells proliferation in a dose-dependent manner of both MCF-7 and HeLa cell lines with the IC50 of 128.6 ± 2.51 and 133.0 ± 1.96 µg PMA equivalents/mL respectively, while the values for the phorbol 12-myristate 13-acetate (PMA) as positive control were 114.7 ± 1.73 and 119.6 ± 3.73 µg/mL, respectively. Microscopic examination showed significant morphological changes that resemble apoptosis in both cell lines when treated with PEs and PMA at IC50 concentration after 24 h. Flow cytometry analysis and DNA fragmentation results confirmed the apoptosis induction of PEs and PMA in both cell lines. The PEs isolated from Jatropha meal activated the PKC-? and down-regulated the proto-oncogenes (c-Myc, c-Fos and c-Jun). These changes probably led to the activation of Caspase-3 protein and apoptosis cell death occurred in MCF-7 and HeLa cell lines upon 24 h treatment with PEs and PMA. Phorbol esters of Jatropha meal were found to be promising as an alternative to replace the chemotherapeutic drugs for cancer therapy.

Ehsan Oskoueian; Norhani Abdullah; Syahida Ahmad

2012-01-01

63

Resistance of cervical adenocarcinoma cells (HeLa) to venom from the scorpion Centruroides limpidus limpidus.  

UK PubMed Central (United Kingdom)

BACKGROUND: The venom of Centruroides limpidus limpidus (Cll) is a mixture of pharmacologically active principles. The most important of these are toxic proteins that interact both selectively and specifically with different cellular targets such as ion channels. Recently, anticancer properties of the venom from other scorpion species have been described. Studies in vitro have shown that scorpion venom induces cell death, inhibits proliferation and triggers the apoptotic pathway in different cancer cell lines. Herein, after treating human cervical adenocarcinoma (HeLa) cells with Cll crude venom, their cytotoxic activity and apoptosis induction were assessed. RESULTS: Cll crude venom induced cell death in normal macrophages in a dose-dependent manner. However, through viability assays, HeLa cells showed high survival rates after exposure to Cll venom. Also, Cll venom did not induce apoptosis after performing ethidium bromide/acridine orange assays, nor was there any evidence of chromatin condensation or DNA fragmentation. CONCLUSIONS: Crude Cll venom exposure was not detrimental to HeLa cell cultures. This may be partially attributable to the absence of specific HeLa cell membrane targets for molecules present in the venom of Centruroides limpidus limpidus. Although these results might discourage additional studies exploring the potential of Cll venom to treat human papilloma cervical cancer, further research is required to explore positive effects of crude Cll venom on other cancer cell lines.

Contreras-Ortiz JM; Vázquez-Chagoyán JC; Martínez-Castañeda JS; Estrada-Franco JG; Aparicio-Burgos JE; Acosta-Dibarrat J; Barbabosa-Pliego A

2013-09-01

64

Initiation of Protein Synthesis in HeLa Cells  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Initiation of protein synthesis in HeLa cells has been synchronized by exposure of the cells to fluoride. Double-labeling of such cells for short pulses with [35S]methionine and a tritiated amino acid, followed by Edman degradation of the puromycin-released nascent peptides, has shown that the perce...

Chatterjee, Nando K.; Kerwar, S. S.; Weissbach, Herbert

65

[Influence of Cx26/Cx32 gap junction channel on antineoplastic effect of etoposide in Hela cells].  

UK PubMed Central (United Kingdom)

OBJECTIVE: To observe the influence of Cx26/Cx32 gap junction channel on the antineoplastic effect of etoposide in Hela cervical cancer cells. METHODS: Fluorescence trace was used to assay the gap junction intercellular communication mediated by Cx26/Cx32 in Hela cells and its functional modulation by the pharmacological agents (oleamide, retinoid acid). A standard colony-forming assay was applied to determine the cell growth-inhibiting effect of etoposide in Hela cells with functional modulation of the gap junction. Hoechst 33258 staining was used to assess the changes in etoposide-induced apoptosis of Hela cells with altered gap junction functions. RESULTS: Oleamide markedly decreased while retinoid acid obviously increased the gap junction function in Hela cells. Standard colony-forming assay showed that etoposide produced a lowered antiproliferative effect in Hela cells with reduced gap junction and an increased antiproliferative effect in cells with enhanced gap junction function. In cells with a reduced gap junction function, etoposide induced a lowered apoptosis rate, which increased obviously in cells with an enhanced gap junction function. CONCLUSION: The antineoplastic effect of etoposide is reduced in Hela cells with a decreased gap junction intercellular communication mediated by Cx26/Cx32 and is enhanced in cells with an increased gap junction intercellular communication.

Tong XH; Dong SY; Jiang GJ; Fan GF

2012-03-01

66

Trichostatin A induces apoptotic cell death of HeLa cells in a Bcl-2 and oxidative stress-dependent manner.  

UK PubMed Central (United Kingdom)

Trichostatin A (TSA) as a HDAC inhibitor can regulate many biological properties including apoptosis and cell proliferation in various cancer cells. Here, we evaluated the effect of TSA on the growth and death of HeLa cervical cancer cells in relation to reactive oxygen species (ROS) and glutathione (GSH) levels. Dose- and time-dependent growth inhibition was observed in HeLa cells with an IC50 of approximately 20 nM at 72 h. This agent also induced apoptotic cell death, as evidenced by annexin V-FITC staining cells, caspase-3 activation and the loss of mitochondrial membrane potential (MMP; ??m). In addition, the administration of Bcl-2 siRNA intensified TSA-induced HeLa cell death. All of the tested caspase inhibitors significantly rescued some cells from TSA-induced HeLa cell death. TSA increased O2•- level and induced GSH depletion in HeLa cells. Caspase inhibitors significantly attenuated O2•- level and GSH depletion in TSA-treated HeLa cells. In addition, N-acetyl cysteine (NAC; a well known antioxidant) significantly prevented cell death and GSH depletion in TSA-treated HeLa cells via decreasing O2•- level. In conclusion, TSA inhibited the growth of HeLa cells via Bcl-2-mediated apoptosis, which was closely related to O2•- and GSH content levels.

You BR; Park WH

2013-01-01

67

Comprehensive profiling of N-linked glycosylation sites in HeLa cells using hydrazide enrichment.  

UK PubMed Central (United Kingdom)

The adenocarcinoma cell line HeLa serves as a model system for cancer research in general and cervical cancer in particular. In this study, hydrazide enrichment in combination with state-of-the art nanoLC-MS/MS analysis was used to profile N-linked glycosites in HeLa cells. N-Linked glycoproteins were selectively enriched in HeLa cells by the hydrazide capture method, which isolates all glycoproteins independent of their glycans. Nonglycosylated proteins were removed by extensive washing. N-Linked glycoproteins were identified with the specific NXT/S motif and deamidated asparagine (N). Deglycosylation was carried out in both H(2)(16)O and H(2)(18)O to confirm the deamidation. NanoLC-MS/MS analysis indicated that the method selectively enriched at least 100 fold N-linked glycosites in HeLa cells. When both the membrane and cytosolic fractions were used, a total of 268 unique N-glycosylation sites were identified corresponding to 106 glycoproteins. Bioinformatic analysis revealed that most of the glycoproteins identified are known to have an impact on cancer and have been proposed as biomarkers.

Malerod H; Graham RL; Sweredoski MJ; Hess S

2013-01-01

68

Antiproliferative and apoptosis inducing activity of Markhamia tomentosa leaf extract on HeLa cells.  

UK PubMed Central (United Kingdom)

ETHNOPHARMACOLOGICAL RELEVANCE: Markhamia tomentosa (Benth) K. Schum ex. Engl. (Bignoniaceae), a tree widely dispersed in West Tropical Africa, is used traditionally to treat various diseases as it possesses antimicrobial, antioxidant, analgesic, anticancer and anti-inflammatory activities. MATERIALS AND METHODS: This study evaluates the cytotoxic effect and underlying mechanisms of the ethanolic extract of Markhamia tomentosa on HeLa and MCF-7 cancer cell lines and non-cancerous Vero cell line. Brine shrimp lethality test was used for preliminary screening. Cytotoxicity was determined using the MTT assay and IC50 was calculated. Effect of Markhamia tomentosa on the cell cycle was monitored by flow cytometry and the apoptosis-induction capability confirmed by exposure of phosphatidylserine to the outer leaflet of the plasma membrane. Loss of mitochondrial membrane potential was analysed by flow cytometry using JC-1. RESULTS: Markhamia tomentosa was toxic to brine shrimps with LD50 of 31.62µg/ml. Cell viability and growth of HeLa cells was inhibited by the extract with an IC50 of 189.1±1.76µg/ml at 24h post treatment. However, no cytotoxic effect was observed in MCF-7 and Vero cell lines. The extract induced cell cycle arrest in HeLa cells in the G0/G1 phase resulting in cell death after 24h exposure. Induction of apoptosis in HeLa cells was substantiated by Annexin V-FITC/PI double staining showing phosphatidylserine translocation and depolarisation of the mitochondrial membrane potential by flow cytometry of JC-1 stained cells. CONCLUSION: The ethanolic extract of Markhamia tomentosa induces G0/G1 in HeLa cells followed by induction of the intrinsic pathway of apoptosis.

Ibrahim B; Sowemimo A; Spies L; Koekomoer T; van de Venter M; Odukoya O

2013-08-01

69

Biocompatibility of Porous Spherical Calcium Carbonate Microparticles on Hela Cells  

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Full Text Available Recently there has been a wide concern on inorganic nanoparticles as drug delivery carriers. CaCO3 particles have shown promising potential for the development of carriers for drugs, but little research had been performed regarding their safe dosage for maximizing the therapeutic activity without harming biosystems. In this study, we assessed the biological safety of porous spherical CaCO3 microparticles on Hela cells. The reactive oxygen species (ROS), glutathione (GSH), carbonyl content in proteins (CCP), DNA-protein crosslinks (DPC) and cell viability were measured. Results showed that with the exposure concentration increase, ROS and CCP in Hela cells presented a significant increase but GSH contents in Hela cells and cell viability showed a significant decrease respectively compared with the control. DPC coefficient ascended, but no statistically significant changes were observed. The results indicated that porous spherical CaCO3 microparticles may induce oxidative damage to Hela cells. But compared with other nanomaterials, porous spherical CaCO3 appeared to have good biocompatibility. The results implied that porous spherical calcium carbonate microparticles could be applied as relatively safe drug vehicles, but with the caveat that the effect of high dosages should not be ignored when attempting to maximize therapeutic activity by increasing the concentration.

Yaran Zhang; Ping Ma; Yao Wang; Juan Du; Qi Zhou; Zhihong Zhu; Xu Yang; Junlin Yuan

2012-01-01

70

In vitro Cytotoxic Activity of ?-chalcogen-substituted Michael-aldol Type Adducts Against Hela and RKO Cell Lines  

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Full Text Available There is a lack of biological studies using selenium and tellurium compounds, specially concern with cancer therapy. The aim of this study is to evaluate the cytotoxicity action of nine ?-chalcogen-substituted Michael-aldol-type adducts on HeLa and RKO cancer cell lines. The cytotoxic effect was assessed by MTT assay and was performed in HeLa and RKO cell lines cultured in RPMI-1640 medium in (95% O2+5% CO2) at 37°C. The IC50 values demonstrated that all compounds presented cytotoxic effect in 17-100 ?M range. The compounds 1 and 5 presented cytotoxic effect in 17 -40 ?M range to HeLa cells. In RKO cells compounds 2-5, 7 and 8 presented cytotoxicity between 46 -58 ?M. Compound 1 presented cytotoxicity for HeLa cells similar to those found to etoposide (11.35±2.73 ?M; p>0.05) and none of the compounds presented this similarity for RKO cells. It is important to notice that the compounds presented cell line selectivity: compound 1 to HeLa and compounds 7 and 8 to RKO cells. The RKO cells were more sensitivity to tellurites, which were not effective to HeLa cells. In conclusion, the compound 1 presented promissory anticancer potential and the cytotoxic specificity of tellurides for RKO cells demonstrated that there is an important biological role based on this chemical element.

Leticia R. Ferreira; Bruno A. Sousa; Fabio V. Santos; Fernando P. Varotti; Alcindo A. Dos Santos; Leandro A. Barbosa

2013-01-01

71

Expression of LGI1 Impairs Proliferation and Survival of HeLa Cells  

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Full Text Available The LGI1 gene was suggested to function as tumor suppressor for its ability to reduce malignant features of glioblastoma cells. In support to this proposal were the findings that overexpression of LGI1 in neuroblastoma cells inhibited proliferation and induced apoptosis. In this study we performed stable LGI1 expression in HeLa cells to examine whether the noxious effect of LGI1 might be extended to cancer cells of diverse origin. HeLa cell clones stably expressing LGI1 exhibited a significant impairment of proliferation and a consistent increase of cell death when compared with control cells lacking expression of LGI1. Expression of LGI1 increased the activity of apoptosis effectors caspase-3/7; furthermore it downregulated the antiapoptotic BCL2 gene and upregulated the proapoptotic BAX gene expression, suggesting that the cause of HeLa cells death might be an increased susceptibility to apoptosis induced by LGI1. The results suggested that LGI1 is capable to restrain growth and survival of adenocarcinoma cells such as HeLa.

Nadia Gabellini; Valentina Masola

2009-01-01

72

Calcium Mobilization in HeLa Cells Induced by Nitric Oxide.  

UK PubMed Central (United Kingdom)

Nitric oxide (NO) has been proposed to be involved in tumor growth and metastasis. However, the mechanism by which nitric oxide modulates cancer cell growth and metastasis on cellular and molecular level is still not fully understood. This work utilized confocal microscopy and fluorescence microplate reader to investigate the effects of exogenous NO on the mobilization of calcium, which is one of the regulators of cell migration, in HeLa cells. The results show that NO elevates calcium in concentration-dependent manner in HeLa cells. And the elevation of calcium induced by NO is due to calcium influx and calcium release from intracellular calcium stores. Moreover, calcium release from intracellular stores is dominant. Furthermore, calcium release from mitochondria is one of the modulation pathways of NO. These findings would contribute to recognizing the significance of NO in cancer cell proliferation and metastasis. SCANNING 9999:XX-XX, 2013. © 2013 Wiley Periodicals, Inc.

Huang Y; Zheng L; Yang H; Chen J; Wang Y; Li H; Xie S

2013-06-01

73

The Effect of Abraxane on Cell Kinetic Parameters of HeLa Cells.  

UK PubMed Central (United Kingdom)

Abraxane (nab-paclitaxel) is a member of the group of nano chemotherapeutics. It is approved for metastatic breast cancer and non small cell lung cancer. Trials for several cancer types including gynecological cancers, head and neck, and prostatic cancer are being studied. In this study, the antiproliferative and apoptotic effect of abraxane was evaluated on HeLa cell line originated from human cervix carcinoma. Three different doses (D1=10 nM, D2=50 nM, D3=100 nM) were administered to HeLa cells for 24, 48 and 72 h. The 50 nM dose of abraxane decreased DNA synthesis from 4.62-0.08%, mitosis from 3.36-1.89% and increased apoptosis from 10.6-30% at 72 h. Additionally, tripolar metaphase plates were seen in mitosis preparations. In this study, abraxane effected cell kinetic parameters significantly. This results are consistent with other studies in the literature.

Gurses N; Topcul M

2013-01-01

74

The Effect of Abraxane on Cell Kinetic Parameters of HeLa Cells.  

Science.gov (United States)

Abraxane (nab-paclitaxel) is a member of the group of nano chemotherapeutics. It is approved for metastatic breast cancer and non small cell lung cancer. Trials for several cancer types including gynecological cancers, head and neck, and prostatic cancer are being studied. In this study, the antiproliferative and apoptotic effect of abraxane was evaluated on HeLa cell line originated from human cervix carcinoma. Three different doses (D1=10 nM, D2=50 nM, D3=100 nM) were administered to HeLa cells for 24, 48 and 72 h. The 50 nM dose of abraxane decreased DNA synthesis from 4.62-0.08%, mitosis from 3.36-1.89% and increased apoptosis from 10.6-30% at 72 h. Additionally, tripolar metaphase plates were seen in mitosis preparations. In this study, abraxane effected cell kinetic parameters significantly. This results are consistent with other studies in the literature. PMID:23991981

Gurses, Nurcan; Topcul, Mehmet

2013-01-01

75

The effect of MAPK inhibitors and ROS modulators on cell growth and death of H?O?-treated HeLa cells.  

UK PubMed Central (United Kingdom)

Reactive oxygen species (ROS) influence the signaling of mitogen?activated protein kinases (MAPKs) involved in cell survival and death. In the present study, the toxicological effect of hydrogen peroxide (H2O2) on HeLa cervical cancer cells was evaluated following treatment with MAPK inhibitors [MAP kinase or ERK kinase (MEK), c?Jun N?terminal kinase (JNK) or p38], N?acetyl cysteine (NAC) and propyl gallate (PG) (well?known antioxidants), or L?buthionine sulfoximine [BSO; an inhibitor of glutathione (GSH) synthesis]. Treatment with 100 µM H2O2 inhibited the growth of HeLa cells and induced cell death, which was accompanied by loss of the mitochondrial membrane potential (MMP; ??m). H2O2 did not induce any specific phase arrests of the cell cycle. ROS levels increased, while GSH levels decreased in H2O2?treated HeLa cells after 1 and 24 h of treatment. The MAPK inhibitors enhanced H2O2?induced HeLa cell death, while only p38 inhibitor increased ROS levels. Both NAC and PG attenuated H2O2?induced HeLa cell growth inhibition and death together with the suppression of ROS levels. BSO increased ROS levels in H2O2?treated HeLa cells without increasing cell death. The levels of MMP (??m) loss and GSH depletion were not closely associated with the levels of apoptosis in HeLa cells treated with the MAPK inhibitors, NAC, PG or BSO, in the presence of H2O2. In conclusion, H2O2 induced HeLa cell growth inhibition and death. MAPK inhibitors generally enhanced H2O2?induced HeLa cell death. In particular, p38 inhibitor increased ROS levels in H2O2?treated HeLa cells, while NAC and PG attenuated H2O2?induced HeLa cell death by suppressing ROS levels.

Park WH

2013-08-01

76

Chemical composition of the essential oil from basil (Ocimum basilicum Linn.) and its in vitro cytotoxicity against HeLa and HEp-2 human cancer cell lines and NIH 3T3 mouse embryonic fibroblasts.  

UK PubMed Central (United Kingdom)

This study examines the chemical composition and in vitro anticancer activity of the essential oil from Ocimum basilicum Linn. (Lamiaceae), cultivated in the Western Ghats of South India. The chemical compositions of basil fresh leaves were identified by GC-MS: 11 components were identified. The major constituents were found to be methyl cinnamate (70.1%), linalool (17.5%), ?-elemene (2.6%) and camphor (1.52%). The results revealed that this plant may belong to the methyl cinnamate and linalool chemotype. A methyl thiazol tetrazolium assay was used for in vitro cytotoxicity screening against the human cervical cancer cell line (HeLa), human laryngeal epithelial carcinoma cell line (HEp-2) and NIH 3T3 mouse embryonic fibroblasts. The IC(50) values obtained were 90.5 and 96.3?µg?mL(-1), respectively, and the results revealed that basil oil has potent cytotoxicity.

Kathirvel P; Ravi S

2012-01-01

77

Chemical composition of the essential oil from basil (Ocimum basilicum Linn.) and its in vitro cytotoxicity against HeLa and HEp-2 human cancer cell lines and NIH 3T3 mouse embryonic fibroblasts.  

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This study examines the chemical composition and in vitro anticancer activity of the essential oil from Ocimum basilicum Linn. (Lamiaceae), cultivated in the Western Ghats of South India. The chemical compositions of basil fresh leaves were identified by GC-MS: 11 components were identified. The major constituents were found to be methyl cinnamate (70.1%), linalool (17.5%), ?-elemene (2.6%) and camphor (1.52%). The results revealed that this plant may belong to the methyl cinnamate and linalool chemotype. A methyl thiazol tetrazolium assay was used for in vitro cytotoxicity screening against the human cervical cancer cell line (HeLa), human laryngeal epithelial carcinoma cell line (HEp-2) and NIH 3T3 mouse embryonic fibroblasts. The IC(50) values obtained were 90.5 and 96.3?µg?mL(-1), respectively, and the results revealed that basil oil has potent cytotoxicity. PMID:21939371

Kathirvel, Poonkodi; Ravi, Subban

2011-09-22

78

Phosphatidylinositol anchor of HeLa cell alkaline phosphatase  

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Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either /sup 3/H-fatty acids or (/sup 3/H)ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the /sup 3/H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of (/sup 3/H)ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from /sup 3/H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the /sup 3/H-fatty acid and the (/sup 3/H)ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the (/sup 3/H)ethanolamine label from the purified alkaline phosphatase. The /sup 3/H radioactivity in alkaline phosphatase purified from (/sup 3/H)ethanolamine-labeled cells comigrated with authentic (/sup 3/H)ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the /sup 3/H-fatty acid and (/sup 3/H)ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase.

Jemmerson, R.; Low, M.G.

1987-09-08

79

Phosphatidylinositol anchor of HeLa cell alkaline phosphatase  

International Nuclear Information System (INIS)

Alkaline phosphatase from cancer cells, HeLa TCRC-1, was biosynthetically labeled with either 3H-fatty acids or [3H]ethanolamine as analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography of immunoprecipitated material. Phosphatidylinositol-specific phospholipase C (PI-PLC) released a substantial proportion of the 3H-fatty acid label from immunoaffinity-purified alkaline phosphatase but had no effect on the radioactivity of [3H]ethanolamine-labeled material. PI-PLC also liberated catalytically active alkaline phosphatase from viable cells, and this could be selectively blocked by monoclonal antibodies to alkaline phosphatase. However, the alkaline phosphatase released from 3H-fatty acid labeled cells by PI-PLC was not radioactive. By contrast, treatment with bromelain removed both the 3H-fatty acid and the [3H]ethanolamine label from purified alkaline phosphatase. Subtilisin was also able to remove the [3H]ethanolamine label from the purified alkaline phosphatase. The 3H radioactivity in alkaline phosphatase purified from [3H]ethanolamine-labeled cells comigrated with authentic [3H]ethanolamine by anion-exchange chromatography after acid hydrolysis. The data suggest that the 3H-fatty acid and [3H]ethanolamine are covalently attached to the carboxyl-terminal segment since bromelain and subtilisin both release alkaline phosphatase from the membrane by cleavage at that end of the polypeptide chain. The data are consistent with findings for other proteins recently shown to be anchored in the membrane through a glycosylphosphatidylinositol structure and indicate that a similar structure contributes to the membrane anchoring of alkaline phosphatase

1987-09-08

80

Kaempferitrin induces apoptosis via intrinsic pathway in HeLa cells and exerts antitumor effects.  

UK PubMed Central (United Kingdom)

ETHNOPHARMACOLOGICAL RELEVANCE: Justicia spicigera is used for the empirical treatment of cervical cancer in Mexico. Recently, we showed that Justicia spicigera extracts exerted cytotoxic and antitumoral effects and the major component of this extract was kaempferitrin (KM). MATERIALS AND METHODS: The cytotoxic and apoptotic effect of KM on human cancer cells and human nontumorigenic cells were evaluated using MTT and TUNEL assays, and Annexin V/Propidium iodide detection by flow cytometry. The effect of KM on cell cycle was analyzed by flow cytometry with propidium iodide. The apoptotic and cell cycle effects were also evaluated by western blot analysis. Also, different doses of KM were injected intraperitoneally daily into athymic mice bearing tumors of HeLa cells during 32 days. The growth and weight of tumors were measured. RESULTS: KM induces high cytotoxic effects in vitro and in vivo against HeLa cells. The general mechanisms by which KM induces cytotoxic effects include: cell cycle arrest in G1 phase and apoptosis via intrinsic pathway in a caspase dependent pathway. Also, KM exerts chemopreventive and antitumor effects. CONCLUSION: KM exerts cytotoxic and antitumor effects against HeLa cells.

Alonso-Castro AJ; Ortiz-Sánchez E; García-Regalado A; Ruiz G; Núñez-Martínez JM; González-Sánchez I; Quintanar-Jurado V; Morales-Sánchez E; Dominguez F; López-Toledo G; Cerbón MA; García-Carrancá A

2013-01-01

 
 
 
 
81

Inhibitory effects and underlying mechanism of 7-hydroxyflavone phosphate ester in HeLa cells.  

UK PubMed Central (United Kingdom)

Chrysin and its phosphate ester have previously been shown to inhibit cell proliferation and induce apoptosis in Hela cells; however, the underlying mechanism remains to be characterized. In the present study, we therefore synthesized diethyl flavon-7-yl phosphate (FP, C(19)H(19)O(6)P) by a simplified Atheron-Todd reaction, and explored its anti-tumor characteristics and mechanisms. Cell proliferation, cell cycle progression and apoptosis were measured by MTS, flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling techniques, respectively in human cervical cancer HeLa cells treated with 7-hydroxyflavone (HF) and FP. p21, proliferating cell nuclear antigen (PCNA) and cAMP levels in Hela cells were analyzed by western blot and radioimmunoassay. Both HF and FP inhibited proliferation and induced apoptosis in HeLa cells via induction of PCNA/p21 expression, cleaved caspase-3/poly (ADP-ribose) polymerase (PARP)-1, elevation of cAMP levels, and cell cycle arrest with accumulation of cells in the G0/G1 fraction. The effects of FP were more potent than those of HF. The interactions of FP with Ca(2+)-calmodulin (CaM) and Ca(2+)-CaM-phosphodiesterase (PDE)1 were explored by electrospray ionization-mass spectrometry and fluorescence spectra. FP, but not HF, formed non-covalent complexes with Ca(2+)-CaM-PDE1, indicating that FP is an inhibitor of PDE1, and resulting in elevated cellular cAMP levels. It is possible that the elevated cAMP levels inhibit growth and induce apoptosis in Hela cells through induction of p21 and cleaved caspase-3/PARP-1 expression, and causing down-regulation of PCNA and cell cycle arrest with accumulation of cells in the G0/G1 and G2/M fractions. In conclusion, FP was shown to be a Ca(2+)-CaM-PDE inhibitor, which might account for its underlying anti-cancer mechanism in HeLa cells. These observations clearly demonstrate the special roles of phosphorylated flavonoids in biological processes, and suggest that FP might represent a potential new drug for the therapy of human cervical carcinoma.

Zhang T; Du J; Liu L; Chen X; Yang F; Jin Q

2012-01-01

82

Inhibitory Effects and Underlying Mechanism of 7-Hydroxyflavone Phosphate Ester in HeLa Cells  

Science.gov (United States)

Chrysin and its phosphate ester have previously been shown to inhibit cell proliferation and induce apoptosis in Hela cells; however, the underlying mechanism remains to be characterized. In the present study, we therefore synthesized diethyl flavon-7-yl phosphate (FP, C19H19O6P) by a simplified Atheron-Todd reaction, and explored its anti-tumor characteristics and mechanisms. Cell proliferation, cell cycle progression and apoptosis were measured by MTS, flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling techniques, respectively in human cervical cancer HeLa cells treated with 7-hydroxyflavone (HF) and FP. p21, proliferating cell nuclear antigen (PCNA) and cAMP levels in Hela cells were analyzed by western blot and radioimmunoassay. Both HF and FP inhibited proliferation and induced apoptosis in HeLa cells via induction of PCNA/p21 expression, cleaved caspase-3/poly (ADP-ribose) polymerase (PARP)-1, elevation of cAMP levels, and cell cycle arrest with accumulation of cells in the G0/G1 fraction. The effects of FP were more potent than those of HF. The interactions of FP with Ca2+-calmodulin (CaM) and Ca2+-CaM-phosphodiesterase (PDE)1 were explored by electrospray ionization-mass spectrometry and fluorescence spectra. FP, but not HF, formed non-covalent complexes with Ca2+-CaM-PDE1, indicating that FP is an inhibitor of PDE1, and resulting in elevated cellular cAMP levels. It is possible that the elevated cAMP levels inhibit growth and induce apoptosis in Hela cells through induction of p21 and cleaved caspase-3/PARP-1 expression, and causing down-regulation of PCNA and cell cycle arrest with accumulation of cells in the G0/G1 and G2/M fractions. In conclusion, FP was shown to be a Ca2+-CaM-PDE inhibitor, which might account for its underlying anti-cancer mechanism in HeLa cells. These observations clearly demonstrate the special roles of phosphorylated flavonoids in biological processes, and suggest that FP might represent a potential new drug for the therapy of human cervical carcinoma.

Liu, Liguo; Chen, Xiaolan; Yang, Fang; Jin, Qi

2012-01-01

83

Transcriptional inactivity of Alu repeats in HeLa cells.  

UK PubMed Central (United Kingdom)

The in vivo transcription of human Alu family members has been investigated by a sensitive primer extension method. The selected primers represent various regions of the Alu family consensus sequence, thus assaying the transcriptional activity of the entire family rather than the activity of an individual member sequence. Using this method, a very small number of RNA molecules per HeLa cell is found to have a distribution of 5' ends centered on the in vitro Alu transcription start site. The distribution of these 5' ends suggests that they are more likely the result of hnRNA degradation rather than transcription start sites. Therefore, despite their great numerical abundance, Alu family members are transcriptionally silent in HeLa cells.

Paulson KE; Schmid CW

1986-08-01

84

Lysosome dysfunction enhances oxidative stress-induced apoptosis through ubiquitinated protein accumulation in Hela cells.  

UK PubMed Central (United Kingdom)

The role of lysosomal system in oxidative stress-induced apoptosis in cancer cells is not fully understood. Menadione is frequently used as oxidative stress model. It is indicated that menadione could induce autophagy in Hela cells. In the present study, we examined whether the lysosomal inhibitor, ammonium chloride (NH(4)Cl) could prevent the autophagy flux by inhibiting the fusion of autophagosomes with lysosomes and enhance apoptosis induced by menadione via mitochondrial pathway. The results demonstrated generation and accumulation of reactive oxygen species and increased levels of ubiquitinated proteins and GRP78 in cells treated with both menadione and NH(4)Cl. Our data indicates that lysosomal system through autophagy plays an important role in preventing menadione-induced apoptosis in Hela cells by clearing misfolded proteins, which alleviates endoplasmic reticulum stress.

Yu C; Huang X; Xu Y; Li H; Su J; Zhong J; Kang J; Liu Y; Sun L

2013-01-01

85

Wogonin and neobaicalein from Scutellaria litwinowii roots are apoptotic for HeLa cells  

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Full Text Available Chemical investigation on the CH2Cl2 fraction of the Scutellaria litwinowii Bornm. & Sint., Lamiaceace, root extract for the first time resulted in the isolation of wogonin, and neobaicalein. These compounds were evaluated for their cytotoxicity towards HeLa cell lines and lymphocytes. Meanwhile, the role of apoptosis was explored in this toxicity. The cells were cultured in RPMI medium and incubated with different concentrations of isolated flavonoids. Cell viability was quantified by MTS assay. Apoptotic cells were determined using propidium iodide staining of DNA fragmentation by flow cytometry (sub-G1peak). Wogonin, and neobaicalein inhibited the growth of malignant cells in a dose-dependent manner. The IC50 values of 46.62 and 79.34 µM were, respectively, found for neobaicalein and wogonin against HeLa cells after 48 h of treatment. Neobaicalein induced a sub-G1 peak in the flow cytometry histogram of treated cells compared to control cells indicating that apoptotic cell death is involved in neobaicalein toxicity. Neobaicalein exerts cytotoxic and pro-apoptotic effects in HeLa cell lines and could be considered as a potential chemotherapeutic agent in cancer treatment.

Zahra Tayarani-Najarani; Javad Asili; Heydar Parsaee; Seyed Hadi Mousavi; Naser Vadati Mashhadian; Alireza Mirzaee; Seyed Ahmad Emami

2012-01-01

86

Preparation of Nuclear Extracts from HeLa Cells.  

UK PubMed Central (United Kingdom)

HeLa cells are the archetypal tissue culture cell line for the preparation of mammalian cell-free systems. These cells, derived from a cervical carcinoma, have been maintained in culture since the late 1940s. They grow with a doubling time of ?24 h and can be cultured in medium containing fetal bovine serum (optimal serum for growth) or medium containing horse serum (in which they grow more slowly, but this serum is considerably less expensive). Nuclear extracts prepared from these cells have been used to determine the mechanisms of splicing and polyadenylation, and such extracts have been characterized extensively. HeLa cells are usually the cell type of choice for initiating cell-free analysis of nearly any aspect of mammalian gene expression. In some instances (e.g., analysis of tissue-specific alternative splicing), it is necessary to use nuclei from a different cell type. We have found that the protocol described here can be used successfully to prepare active nuclear extracts from a wide variety of tissue culture cells, including Drosophila S2 cells.

Nilsen TW

2013-06-01

87

Lycium barbarum polysaccharide inhibits the proliferation of HeLa cells by inducing apoptosis.  

Science.gov (United States)

BACKGROUND: Lycium barbarum polysaccharide (LBP), isolated with boiling water from the famous Chinese medicinal herb Lycium barbarum fruits, is one of the most important functional constituents in Lycium barbarum. In this study the effects of LBP on cell proliferation, cell cycle and apoptosis in human cervical carcinoma cells (HeLa cells) were investigated. RESULTS: LBP could inhibit the proliferation of HeLa cells by changing cell cycle distribution and inducing apoptosis. In addition, the loss of mitochondrial transmembrane potential (??(m) ) was observed by flow cytometry and the increase of intracellular Ca(2+) concentration was detected by laser scanning confocal microscope in apoptotic cells. At the same time, the nitric oxide content, nitric oxide synthase and inducible nitric oxide synthase activities were also increased. CONCLUSION: The inhibitory effect of LBP on the proliferation of HeLa cells was caused by inducing apoptosis through the mitochondrial pathway. The results showed that LBP can be developed as a potential chemotherapeutic agent candidate against human cervical cancer. Copyright © 2012 Society of Chemical Industry. PMID:22696075

Zhu, Cai-Ping; Zhang, Sheng-Hua

2012-06-13

88

Lycium barbarum polysaccharide inhibits the proliferation of HeLa cells by inducing apoptosis.  

UK PubMed Central (United Kingdom)

BACKGROUND: Lycium barbarum polysaccharide (LBP), isolated with boiling water from the famous Chinese medicinal herb Lycium barbarum fruits, is one of the most important functional constituents in Lycium barbarum. In this study the effects of LBP on cell proliferation, cell cycle and apoptosis in human cervical carcinoma cells (HeLa cells) were investigated. RESULTS: LBP could inhibit the proliferation of HeLa cells by changing cell cycle distribution and inducing apoptosis. In addition, the loss of mitochondrial transmembrane potential (??(m) ) was observed by flow cytometry and the increase of intracellular Ca(2+) concentration was detected by laser scanning confocal microscope in apoptotic cells. At the same time, the nitric oxide content, nitric oxide synthase and inducible nitric oxide synthase activities were also increased. CONCLUSION: The inhibitory effect of LBP on the proliferation of HeLa cells was caused by inducing apoptosis through the mitochondrial pathway. The results showed that LBP can be developed as a potential chemotherapeutic agent candidate against human cervical cancer. Copyright © 2012 Society of Chemical Industry.

Zhu CP; Zhang SH

2012-06-01

89

Cloning of smac gene and its overexpression effects on radiosensitivity of HeLa cells to ?-rays  

International Nuclear Information System (INIS)

Objective: To clone smac gene and construct eukaryocytic expression vector pcDNA3.1/ smac. The smac gene was transfected into HeLa cells to explore the effects of over-expression of extrinsic smac gene on radiosensitivity to ?-rays of HeLa cells. Methods: The full-length smac gene was amplified from total RNA of HeLa cells by RTPCR. The RTPCR product was ligated with the vector pcDNA3.1 and sequenced. The correct pcDNA3.1/smac was transfected into HeLa cells. The expression of smac gene was tested by RTPCR and Western blot. The cellular growth inhibition rates were evaluated by MTT 48 horns after irradiation with different doses of ?-rays. Results: Recombinant eukaryocytic expression vector pcDNA3.1/smac was successfully constructed. RTPCR and Western blot results indicated that the expression of smac gene of HeLa/smac cells was significantly enhanced compared with the expression of smac gene of HeLa/pcDNA3.1 and HeLa cells. 48 hours after different doses of ?-ray irradiation was significantly higher in pcDNA3.1/smac transfected HeLa/smac cells than those of non-transfected HeLa cells or pcDNA3.1 transfected HeLa/pcDNA3.1 cells, inhabitation rates were 38.85%, 17.64% and 20.32%, respectively. Conclusions: smac gene was successfully cloned. Extrinsic smac gene over-expression could significantly enhance radiosensitivity to ?-ray of HeLa cells, which would herald a new approach to improve radiosensitivity of cervical cancer. (authors)

2006-01-01

90

Liquiritigenin inhibits tumor growth and vascularization in a mouse model of HeLa cells.  

UK PubMed Central (United Kingdom)

Angiogenesis is one of the crucial steps in the transition of a tumor from a small, harmless cluster of mutated cells to a large, malignant growth, capable of spreading to other organs throughout the body. Vascular endothelial growth factor (VEGF) that stimulates vasculogenesis and angiogenesis is thought to be as an anti-angiogenic target for cancer therapy. Liquiritigenin (LQ), a flavanone existing in Radix glycyrrhiza, shows extensive biological activities, such as anti-inflammatory and anti-cancer properties. In our studies, liquiritigenin effectively inhibited the growth of tumors xenografted in nude mice from human cervical cancer cell line HeLa cells, and microvascular density (MVD) of the tumor exposed to liquiritigenin was reduced in a dose dependent manner, especially in the high dose group. Moreover, the expression and secretion of VEGF were down-regulated by the drug in vivo and in vitro. Therefore, liquiritigenin can be further studied on cancer and other diseases associated with VEGF up-regulation.

Liu Y; Xie S; Wang Y; Luo K; Wang Y; Cai Y

2012-01-01

91

Ascorbic acid: effects on ricin intoxicated HeLa cells  

International Nuclear Information System (INIS)

A study of ricin was made to acertain if ascorbic acid had a specific effect on diphteria toxin or could it prevent the action of toxins from various sources with an activity different than that of diphteria. Ricin was isolated by suspending the defatted meal in double distilled water and adjusting to pH 3.8. The suspension was filtered, the precipitate collected and again dissolved in double distilled water. After saturation with ammonium sulfate, precipitate was collected by centrifugation. The concentration of ricin needed to inhibit at least 50% of the incorporation of (14C) alanine into trichloroacetic acid (TCA) precipitable material was determined. HeLa cells are protected by using ascorbic acid. Ascorbic acid or citric acid was added to the medium 30 min prior to the addition of toxic protein. The isolated ricin prevented the incorporation of (14C) alanine into TCA precipitate material in HeLa cells at levels of 11.5 to 0.00115 microgram of the toxin per ml of culture media. The addition of 100 microgram of ascorbic acid to the HeLa cell cultures 30 min prior to the addition of ricin completely prevented the inhibition of protein synthesis by ricin. Lesser amounts of ascorbic acid offered less protection. Although these data do not elucidate the mechanism of action of ascorbic acid, they show that in vitro ascorbic acid can prevent the action of this poisonous toxin. The data support the use of pharmacological doses of ascorbic acid in the treatment of various cases of poisoning. (Iwakiri, K.)

1977-01-01

92

Yuk-Hap-Tang induces apoptosis by intervening mn-SOD in human cervical carcinoma HeLa cells.  

UK PubMed Central (United Kingdom)

Yuk-Hap-Tang (YHT) induces cell death in human cervical carcinoma HeLa cells. Caspase-3, -6 and -9 were markedly activated in HeLa cells treated with YHT. The preferred substrate for caspase-3 cysteine protease, PARP, was cleaved to its 85-kDa cleavage product. YHT increased the amount of the anti-apoptotic protein, Bcl-2, and the pro-apoptotic protein, Bax. Although p53 has been reported to accumulate in cancer cells in response to anticancer agents, the p53 expression level was not changed in HeLa cells treated with YHT. Manganese (Mn)-TBAP, a mitochondria-specific SOD mimetic agent and NAC/GSH (N-acetyl cysteine/ reduced glutathione) reduced the YHT-induced cytotoxicity and decreased the number of the YHT-induced apoptotic cells. Furthermore, YHT reduced the expression of Mn-SOD protein and its activity in HeLa cells. The data demonstrate that YHT induces the apoptosis of human cervical carcinoma HeLa cells by intervening Mn-SOD.

Chae HJ; Park JM; Lee GY; Park HR; Chae SW; Jeong GS; Kim HM; Kim SB; Yoo SK; Kim HR

2004-01-01

93

AFM-detected apoptotic changes in morphology and biophysical property caused by paclitaxel in Ishikawa and HeLa cells.  

UK PubMed Central (United Kingdom)

The apoptosis of cancer cells is associated with changes in the important cell properties including morphology, surface roughness and stiffness. Therefore, the changes in morphology and biophysical properties can be a good way of evaluating the anticancer activity of a drug. This study examined the effect of paclitaxel on the properties of Ishikawa and HeLa cells using atomic force microscopy (AFM), and the relationship between the changes in morphology and the biophysical properties and apoptosis was discussed. The viability and proliferation of the cells were analyzed using the methylthiazol tetrazolium (MTT) method and a TUNEL assay to confirm cellular apoptosis due to a paclitaxel treatment. AFM observations clearly showed the apoptotic morphological and biophysical changes in Ishikawa and HeLa cells. After the paclitaxel treatment, the cell membrane was torn and holed, the surface roughness was increased, and the stiffness was decreased. These changes were observed more apparently after a 24 h treatment and in Ishikawa cells compared to HeLa cells. The MTT and TUNEL assays results revealed the Ishikawa cells to be more sensitive to paclitaxel than HeLa cells and definite apoptosis occurred after a 24 h treatment. These results showed good agreement with the AFM results. Therefore, research on the morphological and biophysical changes by AFM in cancer cells will help to evaluate the anticancer activities of the drugs.

Kim KS; Cho CH; Park EK; Jung MH; Yoon KS; Park HK

2012-01-01

94

Intense picosecond pulsed electric fields inhibit proliferation and induce apoptosis of HeLa cells.  

UK PubMed Central (United Kingdom)

A picosecond pulsed electric field (psPEF) is a localized physical therapy for tumors that has been developed in recent years, and that may in the future be utilized as a targeted non?invasive treatment. However, there are limited studies regarding the biological effects of psPEF on cells. Electric field amplitude and pulse number are the main parameters of psPEF that influence its biological effects. In this study, we exposed HeLa cells to a psPEF with a variety of electric field amplitudes, from 100 to 600 kV/cm, and various pulse numbers, from 1,000 to 3,000. An MTT assay was used to detect the growth inhibition, while flow cytometry was used to determine the occurrence of apoptosis and the cell cycle of the HeLa cells following treatment. The morphological changes during cell apoptosis were observed using transmission electron microscopy (TEM). The results demonstrated that the cell growth inhibition rate gradually increased, in correlation with the increasing electric field amplitude and pulse number, and achieved a plateau of maximum cell inhibition 12 h following the pulses. In addition, typical characteristics of HeLa cell apoptosis in the experimental groups were observed by TEM. The results demonstrated that the rate of apoptosis in the experimental groups was significantly elevated in comparison with the untreated group. In the treatment groups, the rate of apoptosis was greater in the higher amplitude groups than in the lower amplitude groups. The same results were obtained when the variable was the pulse number. Flow cytometric analysis indicated that the cell cycle of the HeLa cells was arrested at the G2/M phase following psPEF treatment. Overall, our results indicated that psPEF inhibited cell proliferation and induced cell apoptosis, and that these effects occurred in a dose-dependent manner. In addition, the results demonstrated that the growth of the HeLa cells was arrested at the G2/M phase following treatment. This study may provide a foundation for further in vivo experiments, and for the potential clinical application of psPEF in the treatment of cervical cancer.

Zhang M; Xiong ZA; Chen WJ; Yao CG; Zhao ZY; Hua YY

2013-06-01

95

Study on effect of artemisinin combined with 60Co ?-ray on DNA damage in HeLa and SiHa cells  

International Nuclear Information System (INIS)

Objective: To investigate the effect of Artemisinin combined with 60Co ?-ray on DNA damage in HeLa and SiHa cells of human cervical cancer. Methods: Cell growth kinetics was evaluated by MTT assay to determine the most appropriate drug concentration. Effects of Artemisinin combined with 60Co ?-ray on DNA damage in HeLa and SiHa cells were detected by single cell gel electrophoresis. Results: With the concentration increased during the effect of Artemisinin, the HeLa and SiHa cells had higher inhibition on cell proliferation. The SCGE showed that:the comet cell analysis indexes (the comet cells ratio, Tail Length, Olive Tail Moment and Tail DNA%) there was no statistic difference in between the artemisinin group and the control group (P>0.05). With radiation in the same dose, the comet cell analysis indexes of Hela cells treated with both artermisinin and exposed to radiation were higher than that only exposed to radiation group(P0.05). Conclusion: Artemisinin can not induce DNA damage in both HeLa and SiHa cells, but it can make irradiated HeLa cells DNA damage to be aggravated and enhance HeLa cells' radiation sensitivity. However, Artemisinin has no radiosensitizing effect on SiHa cells. (authors)

2011-01-01

96

Genistein promotes cell death of ethanol-stressed HeLa cells through the continuation of apoptosis or secondary necrosis.  

UK PubMed Central (United Kingdom)

BACKGROUND: Apoptosis is a major target and treatment effect of multiple chemotherapeutical agents in cancer. A soybean isoflavone, genistein, is a well-studied chemopreventive agent and has been reported to potentiate the anticancer effect of some chemotherapeutics. However, its mechanistic basis of chemo-enhancement effect remains to be fully elucidated. METHODS: Apoptotic features of low concentration stressed cancer cells were studied by microscopic method, western blot, immunostaining and annexin V/PI assay. Genistein's effects on unstressed cells and recovering cells were investigated using MTT cell viability assay and LDH cytotoxicity assay. Quantitative real-time PCR was employed to analyze the possible gene targets involved in the recovery and genistein's effect. RESULTS: Low-concentration ethanol stressed cancer cells showed apoptotic features and could recover after stress removal. In stressed cells, genistein at sub-toxic dosage promoted the cell death. Quantitative real-time PCR revealed the up-regulation of anti-apoptotic genes MDM2 and XIAP during the recovery process in HeLa cells, and genistein treatment suppressed their expression. The application of genistein, MDM2 inhibitor and XIAP inhibitor to the recovering HeLa cells caused persistent caspase activity and enhanced cell death. Flow cytometry study indicated that genistein treatment could lead to persistent phosphatidylserine (PS) externalization and necrotic events in the recovering HeLa cells. Caspase activity inhibition shifted the major effect of genistein to necrosis. CONCLUSIONS: These results suggested two possible mechanisms through which genistein promoted cell death in stressed cancer cells. Genistein could maintain the existing apoptotic signal to enhance apoptotic cell death. It could also disrupt the recovering process in caspase-independent manner, which lead to necrotic events. These effects may be related to the enhanced antitumor effect of chemotherapeutic drugs when they were combined with genistein.

Xie X; Wang SS; Wong TC; Fung MC

2013-01-01

97

Formononetin potentiates epirubicin-induced apoptosis via ROS production in HeLa cells in vitro.  

Science.gov (United States)

The frequent development of multidrug resistance (MDR) hampers the efficacy of available anticancer drugs in treating cervical cancer. In this study, we aimed to use formononetin (7-hydroxy-4'-methoxyisoflavone), a potential herbal isoflavone, to intensify the chemosensitivity of human cervical cancer HeLa cells to epirubicin, an anticancer drug. The reactive oxygen species (ROS) levels were correlated with MDR modulation mechanisms, including the transporter inhibition and apoptosis induction. Our results revealed that formononetin significantly enhanced the cytotoxicity of epirubicin. Co-incubation of epirubicin with formononetin increased the ROS levels, including hydrogen peroxide and superoxide free radicals. Epirubicin alone markedly increased the mRNA expression of MDR1, MDR-associated protein (MRP) 1, and MRP2. In contrast, formononetin alone or combined treatment decreased the mRNA expression of MRP1 and MRP2. This result indicates that efflux transporter-mediated epirubicin resistance is inhibited at different degrees by the addition of formononetin. This isoflavone significantly intensified epirubicin uptake into HeLa cells. Apoptosis was induced by formononetin and/or epirubicin, as signified by nuclear DNA fragmentation, chromatin condensation, increased sub-G1 and G2/M phases. The cotreatment triggered the mitochondrial apoptotic pathway indicated by increased Bax-to-Bcl-2 expression ratio, loss of mitochondrial membrane potential, and significant activation of caspase-9 and -3. In addition, extrinsic/caspases-8 apoptotic pathway was also induced by the cotreatment. N-acetyl cysteine abrogated these events induced by formononetin, supporting the involvement of ROS in the MDR reversal mechanism. This study pioneered in demonstrating that formononetin may potentiate the cytotoxicity of epirubicin in HeLa cells through the ROS-mediated MRP inhibition and concurrent activation of the mitochondrial and death receptor pathways of apoptosis. Hence, the circumvention of pump and non-pump resistance using formononetin and epirubicin may pave the way for a powerful chemotherapeutic regimen for treating human cervical cancer. PMID:23867903

Lo, Yu-Li; Wang, Wanjen

2013-07-16

98

Formononetin potentiates epirubicin-induced apoptosis via ROS production in HeLa cells in vitro.  

UK PubMed Central (United Kingdom)

The frequent development of multidrug resistance (MDR) hampers the efficacy of available anticancer drugs in treating cervical cancer. In this study, we aimed to use formononetin (7-hydroxy-4'-methoxyisoflavone), a potential herbal isoflavone, to intensify the chemosensitivity of human cervical cancer HeLa cells to epirubicin, an anticancer drug. The reactive oxygen species (ROS) levels were correlated with MDR modulation mechanisms, including the transporter inhibition and apoptosis induction. Our results revealed that formononetin significantly enhanced the cytotoxicity of epirubicin. Co-incubation of epirubicin with formononetin increased the ROS levels, including hydrogen peroxide and superoxide free radicals. Epirubicin alone markedly increased the mRNA expression of MDR1, MDR-associated protein (MRP) 1, and MRP2. In contrast, formononetin alone or combined treatment decreased the mRNA expression of MRP1 and MRP2. This result indicates that efflux transporter-mediated epirubicin resistance is inhibited at different degrees by the addition of formononetin. This isoflavone significantly intensified epirubicin uptake into HeLa cells. Apoptosis was induced by formononetin and/or epirubicin, as signified by nuclear DNA fragmentation, chromatin condensation, increased sub-G1 and G2/M phases. The cotreatment triggered the mitochondrial apoptotic pathway indicated by increased Bax-to-Bcl-2 expression ratio, loss of mitochondrial membrane potential, and significant activation of caspase-9 and -3. In addition, extrinsic/caspases-8 apoptotic pathway was also induced by the cotreatment. N-acetyl cysteine abrogated these events induced by formononetin, supporting the involvement of ROS in the MDR reversal mechanism. This study pioneered in demonstrating that formononetin may potentiate the cytotoxicity of epirubicin in HeLa cells through the ROS-mediated MRP inhibition and concurrent activation of the mitochondrial and death receptor pathways of apoptosis. Hence, the circumvention of pump and non-pump resistance using formononetin and epirubicin may pave the way for a powerful chemotherapeutic regimen for treating human cervical cancer.

Lo YL; Wang W

2013-10-01

99

Potential antitumor agent from the endophytic fungus Pestalotiopsis photiniae induces apoptosis via the mitochondrial pathway in HeLa cells.  

UK PubMed Central (United Kingdom)

4-(3',3'-Dimethylallyloxy)-5-methyl-6-methoxy-phthalide (DMMP) has previously been isolated from the endophytic fungus Pestalotiopsis photiniae. Although the cytotoxic activities of DMMP have been reported, little is known concerning the molecular mechanism of its cytotoxic effect. In the present study, we investigated the effect of DMMP on the growth of several types of cancer cell lines and investigated the mechanism of its antiproliferative effect. DMMP caused the growth inhibition of human cancer lines HeLa, MCF7 and MDA-MB-231, but had little antiproliferative effect on MRC5 normal lung cells. DMMP also significantly caused cell cycle arrest in the G1 phase and upregulated the cyclin-dependent kinase inhibitor p27KIPI protein in the HeLa cells. Moreover DMMP was able to induce marked nuclear apoptotic morphology in HeLa cells. DMMP induced apoptosis and loss of mitochondrial membrane potential (??m) in the HeLa cells. Although the activated forms of caspase-9 and -3 in HeLa cells were detected, pretreatment with caspase inhibitors (Ac-DEVD-CHO and Z-VAD-FMK) failed to attenuate DMMP-induced cell death. In addition, protein levels of the p53 family members, p53 and p73, were upregulated, and DMMP significantly increased the mRNA expression of pro-apoptotic Bcl-2 family genes (PUMA, NOXA, Bax, Bad and Bim). HPV E6-E7 mRNA levels were reduced. In conclusion, DMMP demonstrates potential for use in the treatment of cervical cancer.

Chen C; Hu SY; Luo DQ; Zhu SY; Zhou CQ

2013-10-01

100

Potential antitumor agent from the endophytic fungus Pestalotiopsis photiniae induces apoptosis via the mitochondrial pathway in HeLa cells.  

Science.gov (United States)

4-(3',3'-Dimethylallyloxy)-5-methyl-6-methoxy-phthalide (DMMP) has previously been isolated from the endophytic fungus Pestalotiopsis photiniae. Although the cytotoxic activities of DMMP have been reported, little is known concerning the molecular mechanism of its cytotoxic effect. In the present study, we investigated the effect of DMMP on the growth of several types of cancer cell lines and investigated the mechanism of its antiproliferative effect. DMMP caused the growth inhibition of human cancer lines HeLa, MCF7 and MDA-MB-231, but had little antiproliferative effect on MRC5 normal lung cells. DMMP also significantly caused cell cycle arrest in the G1 phase and upregulated the cyclin-dependent kinase inhibitor p27KIPI protein in the HeLa cells. Moreover DMMP was able to induce marked nuclear apoptotic morphology in HeLa cells. DMMP induced apoptosis and loss of mitochondrial membrane potential (??m) in the HeLa cells. Although the activated forms of caspase-9 and -3 in HeLa cells were detected, pretreatment with caspase inhibitors (Ac-DEVD-CHO and Z-VAD-FMK) failed to attenuate DMMP-induced cell death. In addition, protein levels of the p53 family members, p53 and p73, were upregulated, and DMMP significantly increased the mRNA expression of pro-apoptotic Bcl-2 family genes (PUMA, NOXA, Bax, Bad and Bim). HPV E6-E7 mRNA levels were reduced. In conclusion, DMMP demonstrates potential for use in the treatment of cervical cancer. PMID:23863966

Chen, Chuan; Hu, Shu-Yuan; Luo, Du-Qiang; Zhu, Si-Yu; Zhou, Chuan-Qi

2013-07-17

 
 
 
 
101

Binding and invasion of HeLa and MRC-5 cells by Streptococcus agalactiae.  

UK PubMed Central (United Kingdom)

The interactions of group B streptococci (GBS) with HeLa cells (an epithelial cell line) and MRC-5 cells (a fibroblastic cell line) were explored. A host-cell invasion assay using GBS strains from all serotypes revealed that GBS invaded HeLa cells to a greater extent than MRC-5 cells. One strain, a serotype V (NCS13), was highly invasive against HeLa cells. All strains were poorly invasive against MRC-5 cells. Further characterization of the binding of NCS13 to HeLa and MRC-5 cell surfaces showed that the lack of recoverable c.f.u. from MRC-5 cells was due to a lack of binding of NCS13 to the MRC-5 cell surface in comparison to HeLa cells. Although fibronectin had been reported to bind to GBS, fibronectin assays showed 2.7-fold more fibronectin on the MRC-5 cell surface in comparison to HeLa cells, suggesting that other extracellular matrix proteins besides fibronectin may be involved in GBS binding. Scanning electron microscopy of NCS13 and HeLa cells over a 6 h time period showed increased numbers of NCS13 on the HeLa cell surface over time until cell death at 6 h. Direct contact of the HeLa cell surface by NCS13 was found to be necessary for cell death to occur. Further scanning electron microscopy studies found that, once GBS are bound to the HeLa cell surface, HeLa cell microvilli entwine the bacteria, which then enter the HeLa cell in a polar fashion. Cytoskeletal actin is involved, as this process is disrupted by cytochalasin D, and recruitment of actin is visible at the site of adherent chains of GBS. Also, the host-cell signalling enzyme, PI 3-kinase, is involved in the GBS internalization process, since the PI 3-kinase inhibitor, wortmannin, inhibited NCS13 invasion of HeLa cells in a dose-dependent manner.

Tyrrell GJ; Kennedy A; Shokoples SE; Sherburne RK

2002-12-01

102

Multidrug-resistant hela cells overexpressing MRP1 exhibit sensitivity to cell killing by hyperthermia: Interactions with etoposide  

International Nuclear Information System (INIS)

Purpose: Multidrug resistance (MDR) remains one of the primary obstacles in cancer chemotherapy and often involves overexpression of drug efflux transporters such as P-glycoprotein and multidrug resistance protein 1 (MRP1). Regional hyperthermia is undergoing clinical investigation in combination with chemotherapy or radiotherapy. This study evaluates whether hyperthermia can reverse MDR mediated by MRP1 in human cervical adenocarcinoma (HeLa) cells. Methods and materials: Cytotoxicity of hyperthermia and/or etoposide was evaluated using sulforhodamine-B in HeLa cells overexpressing MRP1 and their drug-sensitive counterparts. Glutathione, glutathione peroxidase (GPx), and glutathione S-transferase (GST) were quantified by spectrophotometry. GST isoenzymes were quantified by immunodetection. Caspase activation was evaluated by fluorometry and chromatin condensation by fluorescence microscopy using Hoechst 33258. Necrosis was determined using propidium iodide. Results: The major finding is that HeLa and HeLaMRP cells are both sensitive to cytotoxicity of hyperthermia (41-45 deg C). Hyperthermia induced activation of caspase 3 and chromatin condensation. Although total levels of cell killing were similar, there was a switch from apoptotic to necrotic cell death in MDR cells. This could be explained by decreased glutathione and GPx in MDR cells. MDR cells also contained very low levels of GST and were resistant to etoposide-induced apoptosis. Hyperthermia caused a modest increase in etoposide-induced apoptosis in HeLa and HeLaMRP cells, which required appropriate heat-drug scheduling. Conclusions: Hyperthermia could be useful in eliminating MDR cells that overexpress MRP1.

2004-12-01

103

Methylene blue-mediated photodynamic therapy induces mitochondria-dependent apoptosis in HeLa cell.  

UK PubMed Central (United Kingdom)

Methylene blue (MB), a widely studied reagent, is investigated in this work for its usage in photodynamic therapy (PDT). PDT has been proved to be highly effective in the treatment of different types of cancers. Previous studies showed MB has both high affinity for mitochondria and high photodynamic efficiency. To elucidate the effects of MB in PDT, we analyzed PDT-induced apoptosis in HeLa cells by introducing different doses of MB into the culture media. Our data showed that MB-mediated PDT triggered intense apoptotic cell death through a series of steps, beginning with photochemical generation of reactive oxygen species. The release of cytochrome c and activation of caspase-3 indicated that MB-PDT-mediated apoptosis in HeLa cells was executed by the mitochondria-dependent apoptotic pathway. Importantly, proteomic studies confirmed that expression levels of several mitochondrial proteins were altered in MB-PDT-induced apoptosis, including TRAP1, mitochondrial elongation factor Tu and peroxiredoxin 3 isoform b. Western blot data showed that phosphorylation of ERK1/2 and PKA were reduced in MB-PDT treated cells, indicating several signal molecules participating in this apoptotic cascade. Moreover, MB-PDT induced an increase in the strength of interaction between Bcl-xL and dephosphorylated Bad. This led to loss of the pro-survival function of Bcl-xL and resulted in mitochondria-mediated apoptosis. This study provides solid evidence of a strong induction by MB-PDT of a mitochondria-dependent apoptosis cascade in HeLa cells.

Lu Y; Jiao R; Chen X; Zhong J; Ji J; Shen P

2008-12-01

104

S-phase specificity of cell killing by docetaxel (Taxotere) in synchronised HeLa cells.  

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Cell viability following short (1 h) contact with paclitaxel or docetaxel was assayed using synchronised HeLa cells. Docetaxel proved almost totally lethal against S-phase cells. Its toxicity was only partial against cells in mitosis, and declined to a minimum with progression to G1. For paclitaxel,...

Hennequin, C.; Giocanti, N.; Favaudon, V.

105

Synergistic Anticancer Activity of 5-Aminolevulinic Acid Photodynamic Therapy in Combination with Low-dose Cisplatin on Hela Cells.  

UK PubMed Central (United Kingdom)

Objective: Photodynamic therapy (PDT ) is a promising modality for the treatment of various tumors. In order to assist in optimizing treatment, we applied 5-ALA/PDT in combination with low-dose cisplatin to evaluate cytotoxicity in Hela cells. Methods: Antiproliferative effects of 5-ALA/PDT and cisplatin, alone and in combination, were assessed using MTT assay. To examine levels of apoptosis, Hela cells treated with 5-ALA/PDT, and combination treatment were assessed with Annexin-V/PI by flow cytometry. To investigate the molecular mechanisms underlying alterations in cell proliferation and apoptosis, Western blot analysis was conducted to determine the expression of p53, p21, Bax and Bcl-2 proteins. Results: MTT assays indicated that combination treatment obviously decreased the viability of Hela cells compared to individual drug treatment. In addition, it was confirmed that exposure of Hela cells to 5-ALA/PDT in combination with low-dose cisplatin resulted in more apoptosis in vitro. Synergistic anticancer activity was related to upregulation p53 expression and alteration in expression of p21, Bcl-2 and Bax. Conclusion: Our findings suggest that administration of 5-ALA/PDT in combination with the low-dose cisplatin may be an effective and feasible therapy for cervical cancer.

Wei XQ; Ma HQ; Liu AH; Zhang YZ

2013-01-01

106

Induction of apoptosis in HeLa cells by chloroform fraction of seed extracts of Nigella sativa  

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Full Text Available Abstract Background Cancer remains one of the most dreaded diseases causing an astonishingly high death rate, second only to cardiac arrest. The fact that conventional and newly emerging treatment procedures like chemotherapy, catalytic therapy, photodynamic therapy and radiotherapy have not succeeded in reverting the outcome of the disease to any drastic extent, has made researchers investigate alternative treatment options. The extensive repertoire of traditional medicinal knowledge systems from various parts of the world are being re-investigated for their healing properties. This study progresses in the direction of identifying component(s) from Nigella sativa with anti cancer acitivity. In the present study we investigated the efficacy of Organic extracts of Nigella sativa seed powder for its clonogenic inhibition and induction of apoptosis in HeLa cancer cell. Results Methanolic, n-Hexane and chloroform extracts of Nigella sativa seedz effectively killed HeLa cells. The IC50 values of methanolic, n-hexane, and chloroform extracts of Nigella sativa were 2.28 ?g/ml, 2.20 ?g/ml and 0.41 ng/ml, respectively. All three extracts induced apoptosis in HeLa cells. Apoptosis was confirmed by DNA fragmentation, western blot and terminal transferase-mediated dUTP-digoxigenin-end labeling (TUNEL) assay. Conclusion Western Blot and TUNEL results suggested that Nigella sativa seed extracts regulated the expression of pro- and anti- apoptotic genes, indicating its possible development as a potential therapeutic agent for cervical cancer upon further investigation.

Shafi Gowhar; Munshi Anjana; Hasan Tarique N; Alshatwi Ali A; Jyothy A; Lei David KY

2009-01-01

107

Leptomycin B increases radiosensitization by trichostain A in HeLa cells  

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Histone deacetylase inhibitors (HDIs) are emerging as potentially useful components of anticancer therapy and their radiosensitizing effects have become evident. Specific HDIs are now available that preferentially inhibit specific HDAC classes; TSA inhibits Class I and II HDACs and SK7041 inhibits Class I HDACs. We tested the differential radiosensitization induced by two different classes of HDIs in HeLa cells. We next tested the hypothesis that p53 expression in cancer cells may influence the susceptibility to HDIs by using pharmacologic modification of the p53 status under an isogenic background. It is interesting that p53 expression in the HeLa cells clearly increased the degree of radiosensitization by TSA compared to that of the class I specific inhibitor SK7041. This suggests that p53 may, in part, be responsible for the mechanistic role for the greater radiosensitization induced by Class I and II inhibitors compared to that of the class I specific inhibitors. Thus, these studies are useful in distinguishing between events mediated solely by the Class I HDACs versus those events involving the other classes of HDACs as well. The anticancer efficacy of targeting Class I and II HDACs in conjunction with radiation therapy, may be further enhanced by the restoration of p53 expression.

Kim, In Ah; Kim, Jin Ho; Shin, Jin Hee [Seoul National University College of Medicine, Seoul (Korea, Republic of)] (and others)

2005-06-15

108

The effects of ascorbic acid on diphtheria toxin and intoxicated HeLa cells.  

UK PubMed Central (United Kingdom)

Ascorbic acid (vitamin C) prevented diphtheria toxin from inhibiting the incorporation of [U-14C]-alanine into trichloroacetic acid precipitable material in HeLa cells. Ascorbic acid did not exhibit an effect on the adenosine diphosphate-ribosylation of amino acyl transferase II nor did it separate fragment A from fragment B in "nicked" toxin. A non-specific reducing agent, para-methylaminophenol sulfate, exhibited an effect on HeLa cells very similar to the results of ascorbic acid. Citric acid, a tricarboxylic acid, had no effect on HeLa cells.

Clark CE; Smith TJ

1976-01-01

109

The products of the reaction between 6-amine-1,3-dimethyl uracil and bis-chalcones induce cytotoxicity with massive vacuolation in HeLa cervical cancer cell line.  

UK PubMed Central (United Kingdom)

As a part of our research in the chemistry of chalcones we have prepared four pyrimidine monoadducts of bis-chalcones through the reaction with 6-amino-1,3-dimethyl uracil. These compounds displayed cytotoxicity with a massive vacuolation in different human cell lines in vitro. Compound 6 was the most cytotoxic inducer of vacuoles, this compound induced G1 phase cell cycle arrest, and their cytotoxicity went without morphological and biochemical evidence of apoptotic cell death in HeLa cells. In addition, our results showed that this vacuole formation does not require de novo protein synthesis and the content vacuolar is acidic. Compound 6 induce necrotic cell death with excessive vacuolation, similar to a process of autophagy. Spautin-1 an inhibitor of autophagy, decreased the transformation of microtubule-associated protein 1 light chain 3 (LC3B-I) to LC3B-II and the vacuolation induced by compound 6 in HeLa cells, both autophagy processes. These compounds could be of pivotal importance in the study of non-apoptotic cell death with vacuole formation and could be useful in research into new autophagy inhibitors agents.

Solano JD; González-Sánchez I; Cerbón MA; Guzmán Á; Martínez-Urbina MA; Vilchis-Reyes MA; Martínez-Zuñiga EC; Alvarado C; Quintero A; Díaz E

2013-02-01

110

Physcion from marine-derived fungus Microsporum sp. induces apoptosis in human cervical carcinoma HeLa cells.  

UK PubMed Central (United Kingdom)

Recently, the relationship between apoptosis and cancer has been emphasized and the induction of apoptosis is recognized as one of the key mechanisms of anti-cancer agents. Marine-derived fungi are valuable sources of structurally diverse bioactive anticancer agents. In the present study, a marine-derived fungus, Microsporum sp. was cultured and an anthraquinone derivative, physcion (11.8mg) was isolated from the culture broth extract (1710mg). Physcion has shown cytotoxic effect on human cervical carcinoma HeLa cells and its apoptosis induction in HeLa cells was investigated by the expressions of p53, p21, Bax, Bcl-2, caspase-9, and caspase-3 proteins. The Western blot analysis has revealed that physcion could significantly induce cell apoptosis through down-regulating of Bcl-2 expression, up-regulating of Bax expression, and activating the caspase-3 pathway. Furthermore, physcion induced the formation of reactive oxygen species (ROS) in HeLa cells. Collectively, these results suggest that physcion could be a potential candidate in the field of anticancer drug discovery against human cervical cancer.

Wijesekara I; Zhang C; Van Ta Q; Vo TS; Li YX; Kim SK

2013-09-01

111

Revolving door action of breast cancer resistance protein (BCRP) facilitates or controls the efflux of flavone glucuronides from UGT1A9-overexpressing HeLa cells.  

UK PubMed Central (United Kingdom)

Cellular production of flavonoid glucuronides requires the action of both UDP-glucuronosyltransferases (UGT) and efflux transporters since glucuronides are too hydrophilic to diffuse across the cellular membrane. We determined the kinetics of efflux of 13 flavonoid glucuronides using the newly developed HeLa-UGT1A9 cells and correlated them with kinetic parameters derived using expressed UGT1A9. The results indicated that, among the seven monohydroxylflavones (HFs), there was moderately good correlation (r(2) ? 0.65) between the fraction metabolized (fmet) derived from HeLa-UGT1A9 cells and CLint derived from the UGT1A9-mediated metabolism. However, there was weak or no correlation between these two parameters for six dihydroxylflavones (DHFs). Furthermore, there was weak or no correlation between various kinetic parameters (Km, Vmax, or CLint) for the efflux and the metabolism regardless of whether we were using seven HFs, six DHFs, or a combination thereof. Instead, the cellular excretion of many flavonoid glucuronides appears to be controlled by the efflux transporter, and the poor affinity of glucuronide to the efflux transporter resulted in major intracellular accumulation of glucuronides to a level that is above the dosing concentration of its aglycone. Hence, the efflux transporters appear to act as the "Revolving Door" to control the cellular excretion of glucuronides. In conclusion, the determination of a flavonoid's susceptibility to glucuronidation must be based on both its susceptibility to glucuronidation by the enzyme and resulting glucuronide's affinity to the relevant efflux transporters, which act as the "Revolving Door(s)" to facilitate or control its removal from the cells.

Wei Y; Wu B; Jiang W; Yin T; Jia X; Basu S; Yang G; Hu M

2013-05-01

112

Antioxidant, anticancer, and apoptosis-inducing effects of Piper extracts in HeLa cells  

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Full Text Available Objective: Cervical cancer is the second most common cancer as well as one of leading cause of cancer-related death for women worldwide. In regards to that issue, focus of this paper will be on popularly used Piperaceae members including Piper betle L, Piper cf fragile Benth, Piper umbellatum L, Piper aduncum L, Piper pellucidum L. This research was conducted to elucidate the antioxidant, anticancer and apoptosis inducing activities of Piperaceae extracts on cervical cancer cells, namely HeLa cell line. Methods: The anticancer activity was determined by inhibiting the proliferation of cells. Apoptosis inducing was determined by inhibiting proliferation cells and by SubG1 flow cytometry. The antioxidant activity is determined by using superoxide dismutase value and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. Results: The highest anticancer activity at 24 h incubation was found for P.pellucidum extract (IC50: 2.85 µg/ml); The anticancer activity at 48 h incubation was more than at 24 h for all extracts. The highest apoptotic activity was found for P.betle (12.5 µg/ml) at both 24 and 48 h incubatio. The highest antioxidant activity was also represented by P.betle extract. Conclusions: All Piperaceae extracts have high anticancer activity; longer incubation increase anticancer activity. P.betle extract has the highest antioxidant property. [J Exp Integr Med 2013; 3(3.000): 225-230

Wahyu Widowati; Laura Wijaya; Teresa L. Wargasetia; Indra Bachtiar; Yelliantty Yelliantty; Dian R. Laksmitawati

2013-01-01

113

Liquiritigenin Inhibits Tumor Growth and Vascularization in a Mouse Model of Hela Cells  

Directory of Open Access Journals (Sweden)

Full Text Available Angiogenesis is one of the crucial steps in the transition of a tumor from a small, harmless cluster of mutated cells to a large, malignant growth, capable of spreading to other organs throughout the body. Vascular endothelial growth factor (VEGF) that stimulates vasculogenesis and angiogenesis is thought to be as an anti-angiogenic target for cancer therapy. Liquiritigenin (LQ), a flavanone existing in Radix glycyrrhiza, shows extensive biological activities, such as anti-inflammatory and anti-cancer properties. In our studies, liquiritigenin effectively inhibited the growth of tumors xenografted in nude mice from human cervical cancer cell line HeLa cells, and microvascular density (MVD) of the tumor exposed to liquiritigenin was reduced in a dose dependent manner, especially in the high dose group. Moreover, the expression and secretion of VEGF were down-regulated by the drug in vivo and in vitro. Therefore, liquiritigenin can be further studied on cancer and other diseases associated with VEGF up-regulation.

Yuxin Liu; Sirou Xie; Yu Wang; Kang Luo; Yang Wang; Yunqing Cai

2012-01-01

114

Liquiritigenin inhibits tumor growth and vascularization in a mouse model of HeLa cells.  

Science.gov (United States)

Angiogenesis is one of the crucial steps in the transition of a tumor from a small, harmless cluster of mutated cells to a large, malignant growth, capable of spreading to other organs throughout the body. Vascular endothelial growth factor (VEGF) that stimulates vasculogenesis and angiogenesis is thought to be as an anti-angiogenic target for cancer therapy. Liquiritigenin (LQ), a flavanone existing in Radix glycyrrhiza, shows extensive biological activities, such as anti-inflammatory and anti-cancer properties. In our studies, liquiritigenin effectively inhibited the growth of tumors xenografted in nude mice from human cervical cancer cell line HeLa cells, and microvascular density (MVD) of the tumor exposed to liquiritigenin was reduced in a dose dependent manner, especially in the high dose group. Moreover, the expression and secretion of VEGF were down-regulated by the drug in vivo and in vitro. Therefore, liquiritigenin can be further studied on cancer and other diseases associated with VEGF up-regulation. PMID:22692244

Liu, Yuxin; Xie, Sirou; Wang, Yu; Luo, Kang; Wang, Yang; Cai, Yunqing

2012-06-12

115

Human recombinant Cripto-1 increases doubling time and reduces proliferation of HeLa cells independent of pro-proliferation pathways.  

UK PubMed Central (United Kingdom)

Human oncofetal protein Cripto-1 (CR-1) is overexpressed in many types of cancers. CR-1 binds to cell surface Glypican-1 to activate Erk1/2 MAPK and Akt pathways leading to cell proliferation. However, we show that treatment with recombinant CR-1 reduces proliferation of HeLa cells by increasing the doubling time without triggering cell death or cell cycle arrest. Using a comparative study with U-87 MG cells, we show that the pro-proliferative pathway of CR-1 is not effective in HeLa cells due to lower expression of Glypican-1. Further we show that treatment with recombinant CR-1 increases PTEN in HeLa cells leading to downregulation of PI3K/Akt pathway. The anti-proliferative effect gets potentiated when the pro-proliferative pathway is blocked.

Das AB; Loying P; Bose B

2012-05-01

116

Human recombinant Cripto-1 increases doubling time and reduces proliferation of HeLa cells independent of pro-proliferation pathways.  

Science.gov (United States)

Human oncofetal protein Cripto-1 (CR-1) is overexpressed in many types of cancers. CR-1 binds to cell surface Glypican-1 to activate Erk1/2 MAPK and Akt pathways leading to cell proliferation. However, we show that treatment with recombinant CR-1 reduces proliferation of HeLa cells by increasing the doubling time without triggering cell death or cell cycle arrest. Using a comparative study with U-87 MG cells, we show that the pro-proliferative pathway of CR-1 is not effective in HeLa cells due to lower expression of Glypican-1. Further we show that treatment with recombinant CR-1 increases PTEN in HeLa cells leading to downregulation of PI3K/Akt pathway. The anti-proliferative effect gets potentiated when the pro-proliferative pathway is blocked. PMID:22182448

Das, Asim Bikas; Loying, Pojul; Bose, Biplab

2011-12-17

117

Effect of tunicamycin on cisplatin induced apoptosis of HeLa cells  

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Full Text Available Objective ?To investigate the effect of endoplasmic reticulum stress (ER stress) in cisplatin-induced apoptosis of human cervical cancer HeLa cells. Methods ?HeLa cells were used as the study object which were divided into four groups: TUNI (5mg/L) group, cisplatin (6mg/L) group, TUNI(5mg/L)+cisplatin(6mg/L) group, and negative control group (no drug treatment). MTT assay was employed to examine the growth status of the cells. Hoechst staining was used to observe the morphological change in the nucleus. Immunoblotting was used to detect the activation of apoptotic proteins, caspase-3 and caspase-4. Indirect immunofluorescence was used to assess the expression of the protein disulfide isomerase (PDI) and phosphorylated histone H2AX (?-H2AX). Results ?MTT assay showed that the growth inhibition rates were 2.65%±2.71%, 19.60%±4.34%, 44.69%±7.07% and 0% in TUNI group, cisplatin group, TUNI+cisplatin group and control group, respectively (P<0.05). Cisplatin showed a significant inhibitory effect on the growth of HeLa cells, and TUNI enhanced the effect of cisplatin. Statistical significance was found between TUNI+cisplatin group and cisplatin group (P<0.05). Hoechst staining showed that the fluorescence of the nucleus in control group was weak and well-distributed. At 12h after treatment, the nuclei in some HeLa cells in cisplatin group and TUNI+cisplatin group diminished in size, thus showing dense hyperfluorescence, and some of them were broken. The proportion of karyorrhexis cells in TUNI+cisplatin group (44.5%±5.1%) was significantly higher than that in cisplatin group (22.7%±3.9%, P<0.05). Immunoblotting showed the expressions of activated caspase-3 and caspase-4 were up-regulated obviously in cisplatin group. Compared to cisplatin group, the expressions of those proteins significantly increased in TUNI+cisplatin group (P<0.05). Indirect immunofluorescence staining showed no PDI expression and weak fluorescence was found in control group. PDI proteins presented in granular form, distributing around the nuclei with strong fluorescence were found in TUNI group and cisplatin group. PDI proteins showed obviously stronger fluorescence in a large proportion of cells in TUNI+cisplatin group, and the fluorescence intensity was obviously higher than that in TUNI group and cisplatin group. No expression of ?-H2AX protein was found in the nucleus in either control group or TUNI group. However, obvious green fluorescence was observed in nuclei of a part of cells in cisplatin group and TUNI+cisplatin group, no obvious difference existed between the two groups. Conclusion ?Heightened ER stress by tunicamycin may increase the apoptosis of HeLa cells induced by cisplatin.

Ye XU; Zhi-xin LI; Hui-ling CAO; Yang YU; Shi-bing LIU; Xiao-jun WANG

2013-01-01

118

Heterofucan from Sargassum filipendula Induces Apoptosis in HeLa Cells  

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Full Text Available Fucan is a term used to denominate a family of sulfated polysaccharides rich in sulfated L-fucose. Heterofucan SF-1.5v was extracted from the brown seaweed Sargassum filipendula by proteolytic digestion followed by sequential acetone precipitation. This fucan showed antiproliferative activity on Hela cells and induced apoptosis. However, SF-1.5v was not able to activate caspases. Moreover, SF-1.5v induced glycogen synthase kinase (GSK) activation, but this protein is not involved in the heterofucan SF-1.5v induced apoptosis mechanism. In addition, ERK, p38, p53, pAKT and NF?B were not affected by the presence of SF-1.5v. We determined that SF-1.5v induces apoptosis in HeLa mainly by mitochondrial release of apoptosis-inducing factor (AIF) into cytosol. In addition, SF-1.5v decreases the expression of anti-apoptotic protein Bcl-2 and increased expression of apoptogenic protein Bax. These results are significant in that they provide a mechanistic framework for further exploring the use of SF-1.5v as a novel chemotherapeutics against human cervical cancer.

Leandro Silva Costa; Cinthia Beatrice Silva Telles; Ruth Medeiros Oliveira; Leonardo Thiago Duarte Barreto Nobre; Nednaldo Dantas-Santos; Rafael Barros Gomes Camara; Mariana Santana Santos Pereira Costa; Jailma Almeida-Lima; Raniere Fagundes Melo-Silveira; Ivan Rui Lopes Albuquerque; Edda Lisboa Leite; Hugo Alexandre Oliveira Rocha

2011-01-01

119

Effect of gelsemium clegans extract on proliferation of hela cell growth and cell cycle  

UK PubMed Central (United Kingdom)

To investigate the effects of Gelsemium elegans Extract on the proliferation of Hela Cell growth and cell cycle. Analysing cell proliferation in the way of XTT and watching it under a Tinelapse Microscope. Testing the apoptosis and proliferation state by using fluo-cytometry. Comprehensive evaluation the effects of Gelsemium elegans Extract on Hela Cell growth and its proliferation. Group B in Gelsemium was obvious. Remarkable retardation on cell proliferation and apoptosis was observed with Gelsemium treated sample (Group B). The effect was dose and time dependent. The cell cycle in Group B has changed significantly. The G_(0)/G_(1) ratio of cultured cell was declined and cells in S phase was founded increased. The proportion of S period cell has risen. But there was not any obvious change in the proportion of G2/M phase cell. Gelsemium elegans Extract can prohibit the prolification of Hela Cell and induce apoptosis. Antitumor effect of Gelsemium elegans Extract is obviously observed. Thus Group B in Gelsemium may prevent the cell transiton form G1 phase to S phase and induce apoptosis during this period.

Ding Jiannong; An Feiyun; Zeng Ming

2005-01-01

120

Vitamin C in Cultured Human (HeLa) Cells: Lack of Effect on DNA Protection and Repair  

Science.gov (United States)

Aims: Dietary antioxidants, including vitamin C, may be in part responsible for the cancer-preventive effects of fruits and vegetables. Human intervention trials with clinical endpoints have failed to confirm their protective effects, and mechanistic studies have given inconsistent results. Our aim was to investigate antioxidant/ pro-oxidant effects of vitamin C at the cellular level. Experimental approach: We have used the comet assay to investigate effects of vitamin C on DNA damage, antioxidant status, and DNA repair, in HeLa (human tumor) cells, and HPLC to measure uptake of vitamin C into cells. Results: Even at concentrations in the medium as high as 200 ?M, vitamin C did not increase the background level of strand breaks or of oxidized purines in nuclear DNA. Vitamin C is taken up by HeLa cells and accumulates to mM levels. Preincubation of cells with vitamin C did not render them resistant to strand breakage induced by H2O2 or to purine oxidation by photosensitizer plus light. Vitamin C had no effect on the rate of repair of strand breaks or oxidized bases by HeLa cells. However, vitamin C at a concentration of less than 1 ?M, or extract from cells preincubated for 6 h with vitamin C, was able to induce damage (strand breaks) in lysed, histone-depleted nuclei (nucleoids). Conclusion: In these cultured human cells, vitamin C displays neither antioxidant nor pro-oxidant properties; nor does it affect DNA strand break or base excision repair.

Azqueta, Amaya; Costa, Solange; Lorenzo, Yolanda; Bastani, Nasser E.; Collins, Andrew R.

2013-01-01

 
 
 
 
121

Vitamin C in cultured human (HeLa) cells: lack of effect on DNA protection and repair.  

UK PubMed Central (United Kingdom)

AIMS: Dietary antioxidants, including vitamin C, may be in part responsible for the cancer-preventive effects of fruits and vegetables. Human intervention trials with clinical endpoints have failed to confirm their protective effects, and mechanistic studies have given inconsistent results. Our aim was to investigate antioxidant/ pro-oxidant effects of vitamin C at the cellular level. EXPERIMENTAL APPROACH: We have used the comet assay to investigate effects of vitamin C on DNA damage, antioxidant status, and DNA repair, in HeLa (human tumor) cells, and HPLC to measure uptake of vitamin C into cells. RESULTS: Even at concentrations in the medium as high as 200 ?M, vitamin C did not increase the background level of strand breaks or of oxidized purines in nuclear DNA. Vitamin C is taken up by HeLa cells and accumulates to mM levels. Preincubation of cells with vitamin C did not render them resistant to strand breakage induced by H2O2 or to purine oxidation by photosensitizer plus light. Vitamin C had no effect on the rate of repair of strand breaks or oxidized bases by HeLa cells. However, vitamin C at a concentration of less than 1 ?M, or extract from cells preincubated for 6 h with vitamin C, was able to induce damage (strand breaks) in lysed, histone-depleted nuclei (nucleoids). CONCLUSION: In these cultured human cells, vitamin C displays neither antioxidant nor pro-oxidant properties; nor does it affect DNA strand break or base excision repair.

Azqueta A; Costa S; Lorenzo Y; Bastani NE; Collins AR

2013-04-01

122

Hyperthermia HeLa cell treatment with silica coated manganese oxide nanoparticles  

CERN Document Server

HeLa tumour cells incubated with ferromagnetic nanoparticles of manganese oxide perovskite La0.56(SrCa)0.22MnO3 were treated with a high frequency alternating magnetic field. The particles were previously coated with silica to improve their biocompatibility. The control assays made with HeLa tumour cells showed that cell survival and growth rate were not affected by the particle internalization in cells, or by the electromagnetic field on cells without nanoparticles. The application of an alternating electromagnetic field to cells incubated with this silica coated manganese oxide induced a significant cellular damage that finally lead to cell death by an apoptotic mechanism.

Villanueva, A; Alonso, JM; Rueda, T; Martínez, A; Crespo, P; Morales, MP; Fernandez, MA Gonzalez; Valdes, J; Rivero, G

2009-01-01

123

Cell death in HeLa cells upon imperatorin and cisplatin treatment Cell death in HeLa cells upon imperatorin and cisplatin treatment  

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Full Text Available There is growing evidence that commonly applied chemotherapy regimens can be improved by introducingnew, specific, active and low side-effect drugs, or by combining substances to obtain the required clinicaleffect. The aim of the present study was to investigate the effects of imperatorin and cisplatin, applied separatelyor in combination, on apoptosis, necrosis and autophagy induction in the human cervical carcinoma cell line(HeLa). Imperatorin appeared to be a potent autophagy inducer, rather than a necrotic or apoptotic one. Incontrast, cisplatin induced mainly apoptosis and necrosis after 6 h and 24 h, while longer incubation resultedonly in necrosis induction. When HeLa cells were incubated with both drugs, autophagy appeared most frequently,although to a smaller extent than that observed after imperatorin administered alone. At the molecularlevel, autophagy was correlated with the presence of the cleaved form of microtubule-associated protein 1 lightchain LC3 — LC3II. It was also accompanied by the inhibition of heat shock proteins Hsp27 and Hsp72 expression.Our results indicate that imperatorin alone, or in combination with cisplatin, is mainly an autopahgy inducerin HeLa cells.There is growing evidence that commonly applied chemotherapy regimens can be improved by introducingnew, specific, active and low side-effect drugs, or by combining substances to obtain the required clinicaleffect. The aim of the present study was to investigate the effects of imperatorin and cisplatin, applied separatelyor in combination, on apoptosis, necrosis and autophagy induction in the human cervical carcinoma cell line(HeLa). Imperatorin appeared to be a potent autophagy inducer, rather than a necrotic or apoptotic one. Incontrast, cisplatin induced mainly apoptosis and necrosis after 6 h and 24 h, while longer incubation resultedonly in necrosis induction. When HeLa cells were incubated with both drugs, autophagy appeared most frequently,although to a smaller extent than that observed after imperatorin administered alone. At the molecularlevel, autophagy was correlated with the presence of the cleaved form of microtubule-associated protein 1 lightchain LC3 — LC3II. It was also accompanied by the inhibition of heat shock proteins Hsp27 and Hsp72 expression.Our results indicate that imperatorin alone, or in combination with cisplatin, is mainly an autopahgy inducerin HeLa cells.

Joanna Jakubowicz-Gil; Roman Paduch; Zofia Ulz; Dorota Badziul; Kazimierz G?owniak; Antoni Gawron

2012-01-01

124

Peripheral blood mononuclear cells inhibit proliferation and promote apoptosis of HeLa cells following stimulation with Bacillus Calmette-Guerin.  

UK PubMed Central (United Kingdom)

Bacillus Calmette-Guerin (BCG) immunotherapy is established as an effective adjuvant intravesical treatment for non-muscle invasive bladder cancer. BCG is also effective in the treatment of Condylomata acuminata caused by low-risk human papilloma virus (HPV). The aim of this study was to determine the efficacy of BCG for the treatment of cervical cancer or HPV high-risk infections. BCG-activated killer (BAK) cells were incubated with a high-risk HPV18-infected cervical cancer cell line, HeLa. The cell cycle distribution and apoptotic index of the HeLa cells were analyzed by flow cytometry. The alterations of HPV-E7, retinoblastoma (RB) and E2F1 levels were detected at the transcriptional and translational levels. The BAK cell cytotoxicity to HeLa cells was 24.08, 14.74 and 6.8% and the natural killer (NK) cell cytotoxicity was 17.62, 10.78 and 5.8% at the E/T ratios of 40:1, 20:1 and 10:1, respectively. The BAK cells significantly induced the apoptosis of HeLa cells to result in an apoptosis level of 24.2% compared with 13.45% by the NK cell treatment at the ratio of 20:1. BAK cells inhibit the proliferation of HeLa cells by G(1)/S cell cycle arrest and this may be associated with the RB/E2F1 pathway. However, G(1)/S arrest and the alteration of RB protein (pRB) and E2F1 levels in the HeLa cells did not show significant differences between the BAK cell- and NK cell-treated groups. HPV-E7 appeared not to be associated with the alteration in cell cycle progression. This study showed that immunotherapy may be a potential treatment for cervical cancer and that BCG immunotherapy may be an alternative and effective method, but further experiments and clinical trials are required to verify this effect.

Lu X; Wu L; Liu Z; Xie L; Wang S

2013-02-01

125

Amphotericin B and Filipin Effects on L and HeLa Cells: Dose Response  

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Amphotericin B (AmB) and filipin effects on L and HeLa cells were compared by monitoring drug-induced potassium leakage from cells, changes in radioactive uridine incorporation into cellular ribonucleic acid, protein leakage from cells, and cell viability. L cells were much more susceptible to both ...

Kotler-Brajtburg, Janina; Medoff, Gerald; Schlessinger, David; Kobayashi, George S.

126

Adjuvant antiproliferative and cytotoxic effect of aloin in irradiated HeLaS3 cells  

International Nuclear Information System (INIS)

Naturally occurring phytoanthracycline, aloin, was used to radiosensitize HeLaS3 human cervix carcinoma cells. The results indicated that the cytotoxic adjuvant effect of aloin was synergistic with IR at all drug concentrations and comparable to the cytotoxicity of 5-10Gy IR alone. Radiosensitization of HeLaS3 cells was achieved by 60?M aloin which reduced IC50 dose of IR from 3.4- to 2Gy. The cell damage by both agents compromised cell capacity to conduct programmed cell death by apoptosis, and led to the synergic cytotoxic cell death by necrosis. (author)

2006-01-01

127

Characterization of the association of radiolabeled bleomycin A2 with HeLa cells  

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The association of (/sup 3/H)bleomycin A2 and Cu(II):(/sup 3/H)bleomycin A2 with HeLa cells has been characterized. Under the conditions of our experiments, approximately 0.1% of the total drug in the medium associates with HeLa cells. Both forms of the drug bind to HeLa cells in a specific and saturable manner, with a Km of 20 microM and a Vmax of 2.5 pmol/min/10(6) cells. Scatchard analysis of the specific binding data demonstrates a single set of high-affinity binding sites. Cytotoxic activities of both forms of the drug are similar, with a 50% lethal dose of 0.5 microM at 48 hr. The specific binding in HeLa cells of either the labeled metal-free drug or its copper complex is reversible by a 100-fold excess of either unlabeled drug. Interaction of the drug with cells is temperature sensitive but is unaffected by metabolic poisons, suggesting that this process is not energy dependent. Isolation of DNA from HeLa cells incubated with the drug indicates that 1 mol of either (/sup 3/H)bleomycin A2 or Cu(II):(/sup 3/H)bleomycin A2 binds per 10(8) nucleotides. Further studies with the radiolabeled drug are required to define precisely the mechanisms involved in bleomycin uptake and compartmentalization within the cell.

Roy, S.N.; Horwitz, S.B.

1984-04-01

128

Inhibition of colony formation of Hela cells by naturally occurring and synthetic agents.  

UK PubMed Central (United Kingdom)

The present study compared the effects of naturally occurring extracts or compound in combination with synthetic selenium compounds on the colony formation and nucleic acid synthesis of cultured human cervical epitheloid carcinoma cells (Hela). Crude extract of bean (Phaseolus vulgaris) or purified lectin from red kidney bean (Phaseolus vulgaris) in combination with selenomethionine were more effective in inhibiting the colony formation of Hela cells than when these cells were treated with these agents alone. Extracts of saffron (Crocus sativus) and selenite have previously been shown to inhibit the colony formation and nucleic acid synthesis by Hela cells in vitro. In the present study we examined the effects of saffron extract in combination with selenite on the colony formation and DNA and RNA synthesis in Hela cells. We found that the treatment of tumor Hela cells with saffron extract in combination with selenite increased the level of inhibition of the colony formation and nucleic acid synthesis in comparison with cells that were treated with only one of these agents. The inhibitory effect of saffron extract in combination with selenite was modified by intracellular sulfhydryl compounds.

Abdullaev FI; Gonzalez de Mejia E

1996-01-01

129

Tumor promoter TPA mimics irradiation effects on the cell cycle of HeLa cells  

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When asynchronous and synchronous HeLa cells were incubated with small doses (10/sup -7/M) of tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA), a variety of transient alterations in the replication cycle were detected within 24 hours by the use of independent methods. Especially, a delayed passage through the S phase and influences on the G2 phase resemble x-ray irradiation effects on cell cultures.

Kinzel, V.; Richards, J.; Stoehr, M.

1980-10-24

130

Effect of HA14-1 on Apoptosis-Regulating Proteins in HeLa Cells.  

UK PubMed Central (United Kingdom)

Overexpression of Bcl-2 has been recognized in various malignancies. Recently, HA14-1, a Bcl-2 antagonist has been identified for its anti-apoptotic effect. However, mode of action of HA14-1 still remains to be elucidated. In present study, we examined HA14-1 binding efficiency with receptor proteins through molecular docking. Cell viability using HeLa cells was evaluated through MTT assay after exposure to different concentration of HA14-1. Moreover, after HA14-1 exposure, expressions of tumor suppressor protein (p53), BH3-only protein (Puma) and apoptosis-associated proteins were analyzed by western blotting. From the results, it was found that HA14-1 occupied all three domains; BH1, BH2 and BH3 within the hydrophobic pocket of Bcl-2. However, HA14-1 occupied only BH1 and BH3 of Bcl-xl, conversely, no such stable bond was observed for Bax and Bak. ARG107 and TYR101 were the amino acid involved in the binding of HA14-1 to Bcl-2 and Bcl-xl respectively. Additionally, decrease in Bcl-2 and Bcl-xl expression along with increase in p53 and Puma expression after exposure to HA14-1 was observed. The results suggested p53 pathway to be the probable mechanism of action for the induction of apoptosis in HeLa cell by down-regulating effect of anti-apoptotic proteins, suggesting that HA14-1 may provide therapeutic potential for human cervical cancer patient. This article is protected by copyright. All rights reserved.

Rehman K; Tariq M; Akash MS; Ginalli Z; Qazi MH

2013-10-01

131

Cytostatic activity of peptide extracts of medicinal plants on transformed A549, H1299, and HeLa Cells.  

UK PubMed Central (United Kingdom)

Biological activity of peptide extracts of medicinal plants was studied on transformed non-small-cell lung carcinoma A549 cells, lung cancer H1299 cells, and cervical cancer HeLa cells at various cell densities. Cell survival and proliferation were evaluated 72 h after treatment with extracts in concentrations of 0.05, 0.25, and 0.5 microg/microl. The cytostatic effect was produced by peptide extracts of Camelia sinesis Kuntze, Inonotus obliquus, and a mixture Inula helenium L., Chelidonium majus L., Equisetum arvense L., and Inonotus obliquus. Peptide extracts of Hypericum perforatum L. and Laurus nobilis L. in the same concentrations had no effects on proliferative activity and growth of tumor cells.

Tepkeeva II; Aushev VN; Zborovskaya IB; Demushkin VP

2009-01-01

132

Cytostatic activity of peptide extracts of medicinal plants on transformed A549, H1299, and HeLa Cells.  

Science.gov (United States)

Biological activity of peptide extracts of medicinal plants was studied on transformed non-small-cell lung carcinoma A549 cells, lung cancer H1299 cells, and cervical cancer HeLa cells at various cell densities. Cell survival and proliferation were evaluated 72 h after treatment with extracts in concentrations of 0.05, 0.25, and 0.5 microg/microl. The cytostatic effect was produced by peptide extracts of Camelia sinesis Kuntze, Inonotus obliquus, and a mixture Inula helenium L., Chelidonium majus L., Equisetum arvense L., and Inonotus obliquus. Peptide extracts of Hypericum perforatum L. and Laurus nobilis L. in the same concentrations had no effects on proliferative activity and growth of tumor cells. PMID:19526129

Tepkeeva, I I; Aushev, V N; Zborovskaya, I B; Demushkin, V P

2009-01-01

133

Nanosecond pulsed electric fields induce poly(ADP-ribose) formation and non-apoptotic cell death in HeLa S3 cells.  

UK PubMed Central (United Kingdom)

Nanosecond pulsed electric fields (nsPEFs) have recently gained attention as effective cancer therapy owing to their potency for cell death induction. Previous studies have shown that apoptosis is a predominant mode of nsPEF-induced cell death in several cell lines, such as Jurkat cells. In this study, we analyzed molecular mechanisms for cell death induced by nsPEFs. When nsPEFs were applied to Jurkat cells, apoptosis was readily induced. Next, we used HeLa S3 cells and analyzed apoptotic events. Contrary to our expectation, nsPEF-exposed HeLa S3 cells exhibited no molecular signs of apoptosis execution. Instead, nsPEFs induced the formation of poly(ADP-ribose) (PAR), a hallmark of necrosis. PAR formation occurred concurrently with a decrease in cell viability, supporting implications of nsPEF-induced PAR formation for cell death. Necrotic PAR formation is known to be catalyzed by poly(ADP-ribose) polymerase-1 (PARP-1), and PARP-1 in apoptotic cells is inactivated by caspase-mediated proteolysis. Consistently, we observed intact and cleaved forms of PARP-1 in nsPEF-exposed and UV-irradiated cells, respectively. Taken together, nsPEFs induce two distinct modes of cell death in a cell type-specific manner, and HeLa S3 cells show PAR-associated non-apoptotic cell death in response to nsPEFs.

Morotomi-Yano K; Akiyama H; Yano K

2013-08-01

134

Identification of a novel short peptide seal specific to CD59 and its effect on HeLa cell growth and apoptosis.  

UK PubMed Central (United Kingdom)

BACKGROUND: In the past, some small peptide ligands identified by phage display technologies have successfully been used in early cancer diagnostics and therapy. In the present study, a novel CD59-binding peptide was identified and its effect on HeLa cell growth and apoptosis was investigated. METHODS: A phage display library was screened yielding a novel short peptide, sp22, that specifically binds to CD59, a protein that shows altered expression in various diseases, including cancer. The effect of ectopic sp22 administration and exogenous sp22 expression on the growth and apoptosis of HeLa cells was assessed. For the latter, we constructed and transfected a sp22-pIRES vector into HeLa cells. RESULTS: Our results show that sp22 peptides can inhibit the level of CD59 mRNA expression, down-regulate Bcl-2 expression, increase Fas and caspase-3 expression, increase the level of cytolysis, and increase the apoptosis of HeLa cells. In contrast, sp22 peptides had no effect on normal human embryonic lung (HEL) cells exhibiting a relatively low CD59 expression level. Compared to untransfected HeLa cells, exogenously sp22 expressing HeLa cells showed a reduced CD59 expression, an increased complement-mediated lysis, a decreased cellular survival ratio, and an increase in apoptotic cells. CONCLUSION: The newly identified sp22 peptide can, in a dose-dependent manner, inhibit CD59 expression. Concomitantly, sp22 can increase complement-mediated lysis and apoptosis signals. This information may be instrumental for the design of novel therapeutic strategies.

Li B; Gao MH; Chu XM; Xu YJ; Yang F

2012-10-01

135

Dynamic behavior of histone H1 microinjected into HeLa cells  

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Histone H1 was purified from bovine thymus and radiolabeled with tritium by reductive methylation or with /sup 125/I using chloramine-T. Red blood cell-mediated microinjection was then used to introduce the labeled H1 molecules into HeLa cells synchronized in S phase. The injected H1 molecules rapidly entered HeLa nuclei, and a number of tests indicate that their association with chromatin was equivalent to that of endogenous histone H1. The injected molecules copurified with HeLa cell nucleosomes, exhibited a half-life of approx.100h, and were hyperphosphorylated at mitosis. When injected HeLa cells were fused with mouse 3T3 fibroblasts < 10% of the labeled H1 molecules migrated to mouse nuclei during the next 48 h. Despite their slow rate of migration between nuclei, the injected H1 molecules were evenly distributed on mouse and human genomes soon after mitosis of HeLa-3T3 heterokaryons. These results suggest that although most histone H1 molecules are stably associated with interphase chromatin, they undergo extensive redistribution after mitosis.

Wu, L.H.; Kuehl, L.; Rechsteiner, M.

1986-01-01

136

MiR-138 downregulates miRNA processing in HeLa cells by targeting RMND5A and decreasing Exportin-5 stability.  

UK PubMed Central (United Kingdom)

MicroRNAs (miRNAs) are a class of non-coding small RNAs that consist of ?22 nt and are involved in several biological processes by regulating target gene expression. MiR-138 has many biological functions and is often downregulated in cancers. Our results showed that overexpression of miR-138 downregulated target RMND5A (required for meiotic nuclear division 5 homolog A) and reduced Exportin-5 stability, which results in decreased levels of pre-miRNA nuclear export in HeLa cells. We also found that miR-138 could significantly inhibit HeLa cell migration by targeting RMND5A. Our study therefore identifies miR-138-RMND5A-Exportin-5 as a previously unknown miRNA processing regulatory pathway in HeLa cells.

Li J; Chen Y; Qin X; Wen J; Ding H; Xia W; Li S; Su X; Wang W; Li H; Zhao Q; Fang T; Qu L; Shao N

2013-09-01

137

Dose-rate effects on the cell cycle and survival of S3 HeLa and V79 cells  

International Nuclear Information System (INIS)

[en] The effects of continuous irradiation at different dose rates on the cell cycle and on cell survival were studied using synchronized S3 HeLa and V79 cells. The minimum dose rate necessary to stop cell division was found to be approximately 23 rad/hr for HeLa cells and 270 rad/hr for V79 cells. For dose rates that stop cell division, cells progress through G1 and S, with a small delay in the S phase, and are blocked in G2. Appreciable mitotic accumulation was observed for HeLa cells at dose rates which stopped cell division. By comparison, much less mitotic accumulation was observed for V79 cells over a range of dose rates from 37 to 270 rad/hr. Minimum mitotic delays for a variety of dose rates were determined for both cell lines. S3 HeLa cells are much more sensitive in this respect than V79 cells; however, it appeared that for higher dose rates the minimum mitotic delay in HeLa cells asymptotically approached a value of about 35 hr. In addition to the qualitative differences observed for the two cell lines in regard to mitotic accumulation, HeLa cells accumulated for prolonged periods in the presence of colcemid while V79 cells were blocked for only a few hours, HeLa cells show a dramatic effect of redistribution of cells into sensitive phases of the cell cycle during exposure, which was reflected in the survival curves at low dose rate. More cell killing per unit dose was observed at 37 than at 74 rad/hr

1979-01-01

138

A COMPARISON BETWEEN HETEROGENEOUS NUCLEAR RNA AND POLYSOMAL MESSENGER RNA IN HELA CELLS BY RNA-DNA HYBRIDIZATION  

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Heterogeneous nuclear RNA (HnRNA) and mRNA from cytoplasmic polyribosomes of HeLa cells have been compared by RNA-DNA hybridization tests. 1 µg of HeLa cell DNA binds 0.05–0.10 µg of either HnRNA or mRNA. In addition, HeLa DNA that is preexposed to unlabeled HnRNA was found to have a reduced capaci...

Soeiro, Ruy; Darnell, James E.

139

Single-walled carbon nanotube interactions with HeLa cells  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract This work concerns exposing cultured human epithelial-like HeLa cells to single-walled carbon nanotubes (SWNTs) dispersed in cell culture media supplemented with serum. First, the as-received CoMoCAT SWNT-containing powder was characterized using scanning electron microscopy and thermal gravimetric analyses. Characterizations of the purified dispersions, termed DM-SWNTs, involved atomic force microscopy, inductively coupled plasma – mass spectrometry, and absorption and Raman spectroscopies. Confocal microRaman spectroscopy was used to demonstrate that DM-SWNTs were taken up by HeLa cells in a time- and temperature-dependent fashion. Transmission electron microscopy revealed SWNT-like material in intracellular vacuoles. The morphologies and growth rates of HeLa cells exposed to DM-SWNTs were statistically similar to control cells over the course of 4 d. Finally, flow cytometry was used to show that the fluorescence from MitoSOX™ Red, a selective indicator of superoxide in mitochondria, was statistically similar in both control cells and cells incubated in DM-SWNTs. The combined results indicate that under our sample preparation protocols and assay conditions, CoMoCAT DM-SWNT dispersions are not inherently cytotoxic to HeLa cells. We conclude with recommendations for improving the accuracy and comparability of carbon nanotube (CNT) cytotoxicity reports.

Yehia Hadi N; Draper Rockford K; Mikoryak Carole; Walker Erin; Bajaj Pooja; Musselman Inga H; Daigrepont Meredith C; Dieckmann Gregg R; Pantano Paul

2007-01-01

140

THE EFFECT OF ETHIDIUM BROMIDE ON MITOCHONDRIAL DNA SYNTHESIS AND MITOCHONDRIAL DNA STRUCTURE IN HELA CELLS  

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The synthesis of mitochondrial DNA (mDNA) in HeLa cells is selectively inhibited by relatively low concentrations of ethidium bromide. After exposure of cells to strongly inhibitory concentrations of the drug, the apparent superhelix density of mDNA is rapidly increased, as judged by its buoyant de...

Leibowitz, Robert D.

 
 
 
 
141

Trypanosoma cruzi trypomastigotes induce cytoskeleton modifications during HeLa cell invasion  

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Full Text Available It has been recently shown that Trypanosoma cruzi trypomastigotes subvert a constitutive membrane repair mechanism to invade HeLa cells. Using a membrane extraction protocol and high-resolution microscopy, the HeLa cytoskeleton and T. cruzi parasites were imaged during the invasion process after 15 min and 45 min. Parasites were initially found under cells and were later observed in the cytoplasm. At later stages, parasite-driven protrusions with parallel filaments were observed, with trypomastigotes at their tips. We conclude that T. cruzi trypomastigotes induce deformations of the cortical actin cytoskeleton shortly after invasion, leading to the formation of pseudopod-like structures.

Maria Cecília Fernandes; Leonardo Rodrigues de Andrade; Norma Windsor Andrews; Renato Arruda Mortara

2011-01-01

142

Effects of Ficus hirta Vahl. (Wuzhimaotao) extracts on growth inhibition of HeLa cells.  

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Ficus hirta Vahl. (Wuzhimaotao) is widely used as a folk medicine by Hakka people in southern China. In order to ascertain if any major fraction can be attributed to have pronounced anticancer effect, extracts of Wuzhimaotao on cytotoxic and apoptosis of HeLa cell lines were evaluated. HeLa cells were cultured and incubated with different concentrations of crude aqueous extracts (CAE), ethyl acetate extracts (EAE), and butyl alcohol extracts (BAE). It showed CAE, EAE, and BAE decreased cell viability on HeLa cells as a dose-dependent manner, and induced a sub-G1 peak in flow cytometry histogram of treated cells compared to the control. Apoptotic cell death is involved in CAE, EAE, and BAE toxicity, with EAE having a significant decrease in G1 population. An over all analysis of results showed that Wuzhimaotao extracts exert antiproliferative action and growth inhibition on HeLa cells through apoptosis induction which indicates its anticancer properties and deserves further investigation. PMID:21435852

Zeng, You-wei; Liu, Xiao-zhen; Lv, Zhen-cheng; Peng, Yong-hong

2011-03-24

143

Interactions of gonococci with HeLa cells: attachment, detachment, replication, penetration, and the role of protein II.  

UK PubMed Central (United Kingdom)

Colony variants of Neisseria gonorrhoeae differ in their interactions with eucaryotic cells. When gonococci were cultivated with HeLa cell monolayers, the opacity phenotype (Op) became increasingly dominant in the subpopulation of organisms which adhered to the HeLa cells. Once bound, Op organisms displayed very low levels of detachment. Adherent Op gonococci exhibited generation times up to threefold greater than cultures containing gonococci in the absence of HeLa cells. In addition, the progeny of adherent Op organisms remained bound to the HeLa cell monolayer. Both piliated (P+) and transparent (Tr) colony types attached to HeLa cells, but their progeny were retained less efficiently. Gonococci bound to HeLa cells were subjected to the bactericidal action of fresh rat serum and approximately 0.5 to 2.5% survived, irrespective of their opacity or piliation phenotype. Incubation with gentamicin resulted in a 10- to 50-fold further reduction in viability. Pretreatment of HeLa cell monolayers with the microfilament-disrupting agent cytochalasin b diminished gonococcal survival in either serum or gentamicin by up to eightfold. In contrast, cytochalasin b treatment did not decrease survival of the commensal organism N. sicca. The data suggest that very few gonococci are completely interiorized and a small proportion of adherent gonococci are partially protected from the soluble-phase environment by HeLa cells.

Bessen D; Gotschlich EC

1986-10-01

144

Interactions of gonococci with HeLa cells: attachment, detachment, replication, penetration, and the role of protein II.  

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Colony variants of Neisseria gonorrhoeae differ in their interactions with eucaryotic cells. When gonococci were cultivated with HeLa cell monolayers, the opacity phenotype (Op) became increasingly dominant in the subpopulation of organisms which adhered to the HeLa cells. Once bound, Op organisms displayed very low levels of detachment. Adherent Op gonococci exhibited generation times up to threefold greater than cultures containing gonococci in the absence of HeLa cells. In addition, the progeny of adherent Op organisms remained bound to the HeLa cell monolayer. Both piliated (P+) and transparent (Tr) colony types attached to HeLa cells, but their progeny were retained less efficiently. Gonococci bound to HeLa cells were subjected to the bactericidal action of fresh rat serum and approximately 0.5 to 2.5% survived, irrespective of their opacity or piliation phenotype. Incubation with gentamicin resulted in a 10- to 50-fold further reduction in viability. Pretreatment of HeLa cell monolayers with the microfilament-disrupting agent cytochalasin b diminished gonococcal survival in either serum or gentamicin by up to eightfold. In contrast, cytochalasin b treatment did not decrease survival of the commensal organism N. sicca. The data suggest that very few gonococci are completely interiorized and a small proportion of adherent gonococci are partially protected from the soluble-phase environment by HeLa cells. PMID:2875950

Bessen, D; Gotschlich, E C

1986-10-01

145

Measles virus-specified polypeptide synthesis in two persistently infected HeLa cell lines.  

UK PubMed Central (United Kingdom)

Measles virus-directed protein synthesis was examined in two HeLa cell lines (K11 and K11A) that are persistently infected with wild-type measles virus. Four viral proteins (H, hemagglutination protein; P, nucleocapsid-associated protein; NP, the major nucleocapsid protein; and M, the matrix protein) were readily detected in both cell lines by immune precipitation of [(35)S]methionine-labeled cell extracts followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, three (H, NP, and M) of the four viral proteins in both K11 and K11A cells differed from the corresponding viral proteins synthesized in HeLa cells acutely infected with the parental wild-type virus. In addition, the M protein from K11A cells migrated significantly more slowly on sodium dodecyl sulfate-polyacrylamide gel electrophoresis than the M protein from K11 cells, and there appeared to be slight differences in the H and NP proteins between these two persistently infected cell lines. The altered viral proteins detected in K11 and K11A cells appeared to be the result of viral mutations rather than changes in the host cell, since virus recovered from these cells directed the synthesis of similar aberrant viral proteins in HeLa cells. Virus recovered from K11 cells and virus recovered from K11A cells were both temperature sensitive and grew more slowly than wild-type virus. HeLa cells infected with virus recovered from K11 cells readily became persistently infected, resembling the original persistently infected K11 cells. Thus, viral mutations are associated with persistent measles virus infections in cell cultures.

Wechsler SL; Rustigian R; Stallcup KC; Byers KB; Winston SH; Fields BN

1979-09-01

146

Vitamin C in Cultured Human (HeLa) Cells: Lack of Effect on DNA Protection and Repair  

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Full Text Available Aims: Dietary antioxidants, including vitamin C, may be in part responsible for the cancer-preventive effects of fruits and vegetables. Human intervention trials with clinical endpoints have failed to confirm their protective effects, and mechanistic studies have given inconsistent results. Our aim was to investigate antioxidant/ pro-oxidant effects of vitamin C at the cellular level. Experimental approach: We have used the comet assay to investigate effects of vitamin C on DNA damage, antioxidant status, and DNA repair, in HeLa (human tumor) cells, and HPLC to measure uptake of vitamin C into cells. Results: Even at concentrations in the medium as high as 200 ?M, vitamin C did not increase the background level of strand breaks or of oxidized purines in nuclear DNA. Vitamin C is taken up by HeLa cells and accumulates to mM levels. Preincubation of cells with vitamin C did not render them resistant to strand breakage induced by H2O2 or to purine oxidation by photosensitizer plus light. Vitamin C had no effect on the rate of repair of strand breaks or oxidized bases by HeLa cells. However, vitamin C at a concentration of less than 1 ?M, or extract from cells preincubated for 6 h with vitamin C, was able to induce damage (strand breaks) in lysed, histone-depleted nuclei (nucleoids). Conclusion: In these cultured human cells, vitamin C displays neither antioxidant nor pro-oxidant properties; nor does it affect DNA strand break or base excision repair.

Amaya Azqueta; Solange Costa; Yolanda Lorenzo; Nasser E. Bastani; Andrew R. Collins

2013-01-01

147

Nucleolar translocalization of GRA10 of Toxoplasma gondii transfectionally expressed in HeLa cells  

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Toxoplasma gondii GRA10 expressed as a GFP-GRA10 fusion protein in HeLa cells moved to the nucleoli within the nucleus rapidly and entirely. GRA10 was concentrated specifically in the dense fibrillar component of the nucleolus morphologically by the overlap of GFP-GRA10 transfection image with IFA i...

Ahn, Hye-Jin; Kim, Sehra; Nam, Ho-Woo

148

Cytotoxicity and apoptotic effects of nickel oxide nanoparticles in cultured HeLa cells  

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The aim of this study was to observe the cytotoxicity and apoptotic effects of nickel oxide nanoparticles on human cervix epithelioid carcinoma cell line (HeLa). Nickel oxide precursors were synthesized by an nickel sulphate-excess urea reaction in boiling aqueous solution. The synthesized NiO nanoparticles (

2010-01-01

149

Invasion of HeLa 229 cells by virulent Bordetella pertussis.  

UK PubMed Central (United Kingdom)

Phase-dependent invasive behavior of Bordetella pertussis was demonstrated by recovery of viable organisms from gentamicin-treated HeLa cell monolayers and by transmission electron microscopy. Several mutants of B. pertussis with Tn5 or Tn5 lac inserted into various vir-regulated genes were evaluated for differences in their invasive abilities. Mutants lacking filamentous hemagglutinin, pertussis toxin, and two as yet uncharacterized vir-regulated products had levels of invasion significantly lower than that of the parent strain BP338. In contrast, invasion by mutants lacking adenylate cyclase toxin was significantly increased compared with that of wild-type B. pertussis. This increase in invasion was eliminated when concentrations of intracellular cyclic 3'-5' AMP were stimulated by treating HeLa cells with cholera toxin or forskolin. Entry of B. pertussis occurred through a microfilament-dependent phagocytic process, as evidenced by the marked reduction in uptake following treatment of HeLa cells with cytochalasin D. Invasion was inhibited with polyclonal anti-B. pertussis and anti-filamentous hemagglutinin antisera. In addition, a monoclonal antibody against lipooligosaccharide A reduced uptake by 65.5%. The preservation of HeLa cell integrity and the limited replication of intracellular bacteria suggest that invasion may represent a means by which B. pertussis evades an active host immune response.

Ewanowich CA; Melton AR; Weiss AA; Sherburne RK; Peppler MS

1989-09-01

150

Invasion of HeLa 229 cells by virulent Bordetella pertussis.  

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Phase-dependent invasive behavior of Bordetella pertussis was demonstrated by recovery of viable organisms from gentamicin-treated HeLa cell monolayers and by transmission electron microscopy. Several mutants of B. pertussis with Tn5 or Tn5 lac inserted into various vir-regulated genes were evaluate...

Ewanowich, C A; Melton, A R; Weiss, A A; Sherburne, R K; Peppler, M S

151

Construction of cell model of silenced Ku80 and the radiobiology change of HeLa cell  

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[en] Objective: To construct the cell model of Ku80 with expression inhibited by siRNA and to explore the role of Ku80 in radiobiology. Methods: Ku80-siRNA expression plasmids were constructed and HeLa cells were transfected with these plasmids by lipofectamine. Western blotting was used to measure the expression of Ku80. After irradiation with 6 MV X ray, cells were collected and analyzed by flow cytometry for apoptosis and cell cycle at 24, 48 and 72 h; The radiobiology parameters of four cell lines were acquired by clone formation array. Results: Three stable transfected cell clones were obtained, and the inhibition rates of Ku80 protein expression of two positive clones were 89.3% and 96.4%; The apoptosis rates of HeLa cells Ku80 inhibited were higher than control cells at 48 and 72 after X ray irradiation (P0.05). HeLa cells of silenced Ku80 had lower SF2 and D0 than control cells, and their SER (sensitization enhancement ratio) based on D10 were 1.315 and 1.365, respectively. Conclusions: The HeLa cell models with Ku80 expression suppressed were successfully established; the inhibition of Ku80 by siRNA could enhance the radiosensitivity of HeLa cells. (authors)

2007-01-01

152

The influence of hyperthermia on pH of lysosomes of cultured Hela cells  

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HeLa cells in the logarithmic phase of growth were heated up to 42-43 deg C in the SHF field for 10 min. Then, after 30. 60 and 120 min pH of lysosomes was determined by neutral red. In 30 min, pH of lysosomes of heated cells decreased down to 5.51+-0.1 against 5.87+-0.07 in intact cells; in 120 min it reached the initial level.

1986-01-01

153

Mitochondria-targeted superoxide dismutase (SOD2) regulates radiation resistance and radiation stress response in HeLa cells  

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Reactive oxygen species (ROS) act as a mediator of ionizing radiation-induced cellular damage. Previous studies have indicated that MnSOD (SOD2) plays a critical role in protection against ionizing radiation in mammalian cells. In this study, we constructed two types of stable HeLa cell lines overexpressing SOD2, HeLa S3/SOD2 and T-REx HeLa/SOD2, to elucidate the mechanisms underlying the protection against radiation by SOD2. SOD2 overexpression in mitochondria enhanced the survival of HeLa S3 and T-REx HeLa cells following ?-irradiation. The levels of ?H2AX significantly decreased in HeLa S3/SOD2 and T-REx HeLa/SOD2 cells compared with those in the control cells. MitoSoxTM Red assays showed that both lines of SOD2-expressing cells showed suppression of the superoxide generation in mitochondria. Furthermore, flow cytometry with a fluorescent probe (2',7'-dichlorofluorescein) revealed that the cellular levels of ROS increased in HeLa S3 cells during post-irradiation incubation, but the increase was markedly attenuated in HeLa S3/SOD2 cells. DNA microarray analysis revealed that, of 47,000 probe sets analyzed, 117 and 166 probes showed more than 2-fold changes after 5.5 Gy of ?-irradiation in control and HeLa S3/SOD2 cells, respectively. Pathway analysis revealed different expression profiles in irradiated control cells and irradiated SOD2-overexpressing cells. These results indicate that SOD2 protects HeLa cells against cellular effects of ?-rays through suppressing oxidative stress in irradiated cells caused by ROS generated in the mitochondria and through regulating the expression of genes which play a critical role in protection against ionizing radiation. (author)

2012-01-01

154

A phthalide derivative isolated from endophytic fungi Pestalotiopsis photiniae induces G1 cell cycle arrest and apoptosis in human HeLa cells  

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Full Text Available Abstract in english MP [4-(3?,3?-dimethylallyloxy)-5-methyl-6-methoxyphthalide] was obtained from liquid culture of Pestalotiopsis photiniae isolated from the Chinese Podocarpaceae plant Podocarpus macrophyllus. MP significantly inhibited the proliferation of HeLa tumor cell lines. After treatment with MP, characteristic apoptotic features such as DNA fragmentation and chromatin condensation were observed in DAPI-stained HeLa cells. Flow cytome (more) try showed that MP induced G1 cell cycle arrest and apoptosis in a dose-dependent manner. Western blotting and real-time reverse transcription-polymerase chain reaction were used to investigate protein and mRNA expression. MP caused significant cell cycle arrest by upregulating the cyclin-dependent kinase inhibitor p27KIP1 protein and p21CIP1 mRNA levels in HeLa cells. The expression of p73 protein was increased after treatment with various MP concentrations. mRNA expression of the cell cycle-related genes, p21CIP1 , p16INK4a and Gadd45?, was significantly upregulated and mRNA levels demonstrated significantly increased translation of p73, JunB, FKHR, and Bim. The results indicate that MP may be a potential treatment for cervical cancer.

Chen, C.; Yang, R.L.

2013-07-01

155

A phthalide derivative isolated from endophytic fungi Pestalotiopsis photiniae induces G1 cell cycle arrest and apoptosis in human HeLa cells  

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Full Text Available Abstract in english MP [4-(3?,3?-dimethylallyloxy)-5-methyl-6-methoxyphthalide] was obtained from liquid culture of Pestalotiopsis photiniae isolated from the Chinese Podocarpaceae plant Podocarpus macrophyllus. MP significantly inhibited the proliferation of HeLa tumor cell lines. After treatment with MP, characteristic apoptotic features such as DNA fragmentation and chromatin condensation were observed in DAPI-stained HeLa cells. Flow cytome (more) try showed that MP induced G1 cell cycle arrest and apoptosis in a dose-dependent manner. Western blotting and real-time reverse transcription-polymerase chain reaction were used to investigate protein and mRNA expression. MP caused significant cell cycle arrest by upregulating the cyclin-dependent kinase inhibitor p27KIP1 protein and p21CIP1 mRNA levels in HeLa cells. The expression of p73 protein was increased after treatment with various MP concentrations. mRNA expression of the cell cycle-related genes, p21CIP1 , p16INK4a and Gadd45?, was significantly upregulated and mRNA levels demonstrated significantly increased translation of p73, JunB, FKHR, and Bim. The results indicate that MP may be a potential treatment for cervical cancer.

Chen, C.; Yang, R.L.

2013-08-01

156

TSPY potentiates cell proliferation and tumorigenesis by promoting cell cycle progression in HeLa and NIH3T3 cells  

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Full Text Available Abstract Background TSPY is a repeated gene mapped to the critical region harboring the gonadoblastoma locus on the Y chromosome (GBY), the only oncogenic locus on this male-specific chromosome. Elevated levels of TSPY have been observed in gonadoblastoma specimens and a variety of other tumor tissues, including testicular germ cell tumors, prostate cancer, melanoma, and liver cancer. TSPY contains a SET/NAP domain that is present in a family of cyclin B and/or histone binding proteins represented by the oncoprotein SET and the nucleosome assembly protein 1 (NAP1), involved in cell cycle regulation and replication. Methods To determine a possible cellular function for TSPY, we manipulated the TSPY expression in HeLa and NIH3T3 cells using the Tet-off system. Cell proliferation, colony formation assays and tumor growth in nude mice were utilized to determine the TSPY effects on cell growth and tumorigenesis. Cell cycle analysis and cell synchronization techniques were used to determine cell cycle profiles. Microarray and RT-PCR were used to investigate gene expression in TSPY expressing cells. Results Our findings suggest that TSPY expression increases cell proliferation in vitro and tumorigenesis in vivo. Ectopic expression of TSPY results in a smaller population of the host cells in the G2/M phase of the cell cycle. Using cell synchronization techniques, we show that TSPY is capable of mediating a rapid transition of the cells through the G2/M phase. Microarray analysis demonstrates that numerous genes involved in the cell cycle and apoptosis are affected by TSPY expression in the HeLa cells. Conclusion These data, taken together, have provided important insights on the probable functions of TSPY in cell cycle progression, cell proliferation, and tumorigenesis.

Oram Shane W; Liu Xing; Lee Tin-Lap; Chan Wai-Yee; Lau Yun-Fai

2006-01-01

157

Induction of the mitochondrial permeability transition mediates the killing of HeLa cells by staurosporine.  

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The role of the mitochondrial permeability transition (MPT) in the killing of HeLa cells by staurosporine (STR) was assessed with the use of bongkrekic acid (BK), an inhibitor of the MPT. BK prevented cell killing as well as biochemical manifestations of the MPT: (a) the loss of the mitochondrial membrane potential (deltapsim); (b) the release of cytochrome c from the intramembranous space to the cytosol; and (c) the release of malate dehydrogenase from the mitochondrial matrix. Stable transfectants that overexpressed Akt were also resistant to cell killing and did not develop an MPT. STR inhibited the phosphorylation of Bad, whereas Bad phosphorylation was preserved in cells that overexpress Akt. In wild-type HeLa cells treated with STR, the content of Bax in the cytosol decreased as that in the mitochondria increased, a result that was again prevented by overexpression of Akt. Bid accumulation in the mitochondria with STR was not affected by overexpression of Akt. The pan-caspase inhibitor Z-Val-Ala-Val-Asp(OMe) fluoromethylketone prevented cell killing bu not induction of the MPT. The data document the central role of the MPT in the killing of HeLa cells by STR. The data are consistent with the hypothesis that induction of the MPT is a consequence of the movement of Bax to the mitochondria. Phosphorylation of Bad prevents Bax translocation. Caspases participate in the events related to cell killing that occur subsequent to induction of the MPT. PMID:11289115

Tafani, M; Minchenko, D A; Serroni, A; Farber, J L

2001-03-15

158

Induction of the mitochondrial permeability transition mediates the killing of HeLa cells by staurosporine.  

UK PubMed Central (United Kingdom)

The role of the mitochondrial permeability transition (MPT) in the killing of HeLa cells by staurosporine (STR) was assessed with the use of bongkrekic acid (BK), an inhibitor of the MPT. BK prevented cell killing as well as biochemical manifestations of the MPT: (a) the loss of the mitochondrial membrane potential (deltapsim); (b) the release of cytochrome c from the intramembranous space to the cytosol; and (c) the release of malate dehydrogenase from the mitochondrial matrix. Stable transfectants that overexpressed Akt were also resistant to cell killing and did not develop an MPT. STR inhibited the phosphorylation of Bad, whereas Bad phosphorylation was preserved in cells that overexpress Akt. In wild-type HeLa cells treated with STR, the content of Bax in the cytosol decreased as that in the mitochondria increased, a result that was again prevented by overexpression of Akt. Bid accumulation in the mitochondria with STR was not affected by overexpression of Akt. The pan-caspase inhibitor Z-Val-Ala-Val-Asp(OMe) fluoromethylketone prevented cell killing bu not induction of the MPT. The data document the central role of the MPT in the killing of HeLa cells by STR. The data are consistent with the hypothesis that induction of the MPT is a consequence of the movement of Bax to the mitochondria. Phosphorylation of Bad prevents Bax translocation. Caspases participate in the events related to cell killing that occur subsequent to induction of the MPT.

Tafani M; Minchenko DA; Serroni A; Farber JL

2001-03-01

159

A class of DNA-binding peptides from wheat bud causes growth inhibition, G2 cell cycle arrest and apoptosis induction in HeLa cells  

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Full Text Available Abstract Background Deproteinized DNA from eukaryotic and prokaryotic cells still contains a low-molecular weight peptidic fraction which can be dissociated by alkalinization of the medium. This fraction inhibits RNA transcription and tumor cell growth. Removal from DNA of normal cells causes amplification of DNA template activity. This effect is lower or absent in several cancer cell lines. Likewise, the amount of active peptides in cancer cell DNA extracts is lower than in DNA preparation of the corresponding normal cells. Such evidence, and their ubiquitous presence, suggests that they are a regulatory, conserved factor involved in the control of normal cell growth and gene expression. Results We report that peptides extracted from wheat bud chromatin induce growth inhibition, G2 arrest and caspase-dependent apoptosis in HeLa cells. The growth rate is decreased in cells treated during the S phase only and it is accompanied by DNA damage and DNA synthesis inhibition. In G2 cells, this treatment induces inactivation of the CDK1-cyclin B1 complex and an increase of active chk1 kinase expression. Conclusion The data indicate that the chromatin peptidic pool inhibits HeLa cell growth by causing defective DNA replication which, in turn, arrests cell cycle progression to mitosis via G2 checkpoint pathway activation.

Mancinelli Loretta; De Angelis Paula M; Annulli Lucia; Padovini Valentina; Elgjo Kjell; Gianfranceschi Gian

2009-01-01

160

[Effect of nitric oxide on HSV-1 infection of HeLa cells and Vero-E6 cells  

UK PubMed Central (United Kingdom)

The effects of nitric oxide(NO) on HSV-1 infection of HeLa cell line and Vero-E6 cell line were investigated in this study. The titers of HSV-1 in supernatants from infected culture of Vero-E6 cells were decreased by a NO donor, sodium nitroprusside(SNP), in a dose-dependent manner, while the control compound potassium ferricyanide (KFC) had no effect. However, SNP had no effect on that of HeLa cells. The results reaffirm the antiviral effect of NO and suggest that the susceptibility of different cell lines to the effect of NO might be varied.

Liu S; Shu M; Xiao Y

1999-01-01

 
 
 
 
161

Deoxyribonucleic acid synthesis, cell cycle progression, and division of Chlamydia-infected HeLa 229 cells.  

UK PubMed Central (United Kingdom)

The fate of lymphogranuloma venereum strain Chlamydia-infected HeLa 229 cells was examined by determining the rate of deoxyribonucleic acid synthesis and the kinetics of entry into and progression through S phase and by time-lapse cinemicrography. At an input multiplicity of 5 or less, Chlamydia-infected cells showed no inhibition of host deoxyribonucleic acid synthesis or cell cycle progression. Cinemicrography showed division of inclusion-containing cells, with one or both daughters receiving chlamydial inclusions. Analysis of the family trees indicated that the generation times of infected HeLa 229 were not altered relative to those of the uninfected cells.

Bose SK; Liebhaber H

1979-06-01

162

Deoxyribonucleic acid synthesis, cell cycle progression, and division of Chlamydia-infected HeLa 229 cells.  

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The fate of lymphogranuloma venereum strain Chlamydia-infected HeLa 229 cells was examined by determining the rate of deoxyribonucleic acid synthesis and the kinetics of entry into and progression through S phase and by time-lapse cinemicrography. At an input multiplicity of 5 or less, Chlamydia-infected cells showed no inhibition of host deoxyribonucleic acid synthesis or cell cycle progression. Cinemicrography showed division of inclusion-containing cells, with one or both daughters receiving chlamydial inclusions. Analysis of the family trees indicated that the generation times of infected HeLa 229 were not altered relative to those of the uninfected cells. PMID:468381

Bose, S K; Liebhaber, H

1979-06-01

163

Lipophilic pro-oligonucleotides are rapidly and efficiently internalized in HeLa cells.  

UK PubMed Central (United Kingdom)

Model t-Bu-SATE pro-dodecathymidines labeled with fluorescein and exhibiting various lipophilicities were evaluated for their uptake by cells in culture. Pro-oligonucleotides with appropriate lipophilicity were found to permeate across the HeLa cell membrane much more extensively than the control phosphorothioate oligo or than the hydrophilic pro-oligos. Fluorescence patterns of internalization were consistent with a diffusion mechanism resulting in the appearance of a uniform cytoplasmic distribution and nuclear accumulation, as confirmed by confocal microscopy.

Vivès E; Dell'Aquila C; Bologna JC; Morvan F; Rayner B; Imbach JL

1999-10-01

164

Disorganization of mitosis in HeLa cells by deuterium oxide.  

UK PubMed Central (United Kingdom)

We assessed, by light and electron microscopy, the influence of deuterium oxide on the dynamics of mitosis and on the morphology of the mitotic apparatus in HeLa cells grown in vitro. A 2-h incubation of HeLa monolayers with low concentrations of D2O (1%-25%) in the medium increased the frequency of multipolar divisions up to 20 times the control level. Substitution of 10% and 25% D2O for H2O induced changes in the proportions of mitotic phases. These changes could be fully reversed to the control pattern after 1 h of recovery in non-deuterated medium. Fifty % D2O strongly inhibited, and 75% D2O blocked the cell cycle before prophase and at (pro-)metaphase. In cells treated with 50% D2O conspicuous morphological changes of the interphase chromatin as well as ultrastructural abnormalities of all mitotic phases were regularly observed. Overall, these results confirmed the antimitotic activity of deuterium oxide and revealed that it could also influence the cell cycle before mitosis. It is suggested that interference with diverse cellular constituents rather than a specific influence on microtubule turnover could be responsible for the disorganization of the cell cycle in HeLa cells by D2O.

Lamprecht J; Schroeter D; Paweletz N

1989-12-01

165

Cytotoxicity and apoptotic effects of nickel oxide nanoparticles in cultured HeLa cells  

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Full Text Available The aim of this study was to observe the cytotoxicity and apoptotic effects of nickel oxide nanoparticles on humancervix epithelioid carcinoma cell line (HeLa). Nickel oxide precursors were synthesized by an nickel sulphate-excess ureareaction in boiling aqueous solution. The synthesized NiO nanoparticles (<200 nm) were investigated by X-ray diffractionanalysis and transmission electron microscopy techniques. For cytotoxicity experiments, HeLa cells were incubated in50-500 ?g/mL NiO for 2, 6, 12 and 16 hours. The viable cells were counted with a haemacytometer using light microscopy.The cytotoxicity was observed low in 50-200 ?g/mL concentration for 16 h, but high in 400-500 ?g/mL concentration for2-6 h. HeLa cells' cytoplasm membrane was lysed and detached from the well surface in 400 ?g/mL concentration NiOnanoparticles. Double staining and M30 immunostaining were performed to quantify the number of apoptotic cells in cultureon the basis of apoptotic cell nuclei scores. The apoptotic effect was observed 20% for 16 h incubation.

Kezban Ada; Mustafa Turk; Serpil Oguztuzun; Murat Kilic; Mehmet Demirel; Nisa Tandogan; Ertan Ersayar; Ozturk Latif

2010-01-01

166

Cytotoxicity and apoptotic effects of nickel oxide nanoparticles in cultured HeLa cells.  

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Full Text Available The aim of this study was to observe the cytotoxicity and apoptotic effects of nickel oxide nanoparticles on human cervix epithelioid carcinoma cell line (HeLa). Nickel oxide precursors were synthesized by an nickel sulphate-excess urea reaction in boiling aqueous solution. The synthesized NiO nanoparticles (<200 nm) were investigated by X-ray diffraction analysis and transmission electron microscopy techniques. For cytotoxicity experiments, HeLa cells were incubated in 50-500 Î?g/mL NiO for 2, 6, 12 and 16 hours. The viable cells were counted with a haemacytometer using light microscopy. The cytotoxicity was observed low in 50-200 Î?g/mL concentration for 16 h, but high in 400-500 Î?g/mL concentration for 2-6 h. HeLa cells' cytoplasm membrane was lysed and detached from the well surface in 400 Î?g/mL concentration NiO nanoparticles. Double staining and M30 immunostaining were performed to quantify the number of apoptotic cells in culture on the basis of apoptotic cell nuclei scores. The apoptotic effect was observed 20% for 16 h incubation.

Kezban Ada; Mustafa Turk; Serpil Oguztuzun; Murat Kilic; Mehmet Demirel; Nisa Tandogan; Ertan Ersayar; Ozturk Latif

2011-01-01

167

Substitued (E)-b-(benzoyl)acrylic acids suppressed survival of neoplastic human HeLa cells  

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Full Text Available The bacteriostatic activity of some of alkyl substituted (E)-b-(benzoyl)acrylic acids was shown earlier. The aim of this study was to investigate the antiproliferative action of 19 alkyl-, or halogeno-, or methoxy-, or acetamido- substituted (E)-b-(benzoyl)acrylic acids, against human cervix carcinoma, HeLa, cells. Target HeLa cells were continuously treated with increasing concentrations of substituted (E)-b-(benzoyl)acrylic acids during two days. The MTT test was used for assessment of the antiproliferative action of this group of compounds. Treatment of HeLa cells with 4-methyl-, 4-fluoro-, 4-chloro-, 4-bromo- and 4-methoxy- derivatives of (E)-b-(benzoyl) acrylic acid leads to the expression of cytostatic activity against HeLa cells (IC50 were in the range from 31-40 µM). Their antiproliferative action was less than that of the basic compound (E)-b-(benzoyl)acrylic acid whose IC50 was 28.5 µM. The 3,4-dimethyl-, 2,4-dimethyl- and 2,5-dimethyl- derivatives as well as the 4-ethyl- and 3,4-dichloro- and 2,4-dichloro-derivatives, have stronger cytostatic activity than the correspoding monosubstituted and parent compound. Their IC50 were 18.5 µM; 17.5 µM; 17.0 mM; 17.5 µM; 22.0 µM and 18 µM, respectively. The 4-iso-propyl- and 4-n-butyl-derivatives exerted higher cytostatic activity than the compounds with a lower number of methylene -CH2- groups in the substitutent. Their IC50 were 14.5 µM and 6.5 µM respectively. The 2,5-di-iso-propyl- and 4-tert-butyl-derivatives expressed the most strong antiproliferative action against the investigated HeLa cells, IC50 being 4.5 µM and 5.5 µM, respectively. The investigated compounds affected the survival of HeLa cells, expressing a strong structure-activity relationship of the Hansch type.

Z. JURANIC; LJ. STEVOVIC; B. DRAKULIC; T. STANOJKOVIC; S. RADULOVIC; I. JURANIC

1999-01-01

168

Coxsackievirus B5 induced apoptosis of HeLa cells: Effects on p53 and SUMO  

International Nuclear Information System (INIS)

[en] Coxsackievirus B5 (CVB5), a human enterovirus of the family Picornaviridae, is a frequent cause of acute and chronic human diseases. The pathogenesis of enteroviral infections is not completely understood, and the fate of the CVB5-infected cell has a pivotal role in this process. We have investigated the CVB5-induced apoptosis of HeLa cells and found that it happens by the intrinsic pathway by a mechanism dependent on the ubiquitin-proteasome system, associated with nuclear aggregation of p53. Striking redistribution of both SUMO and UBC9 was noted at 4 h post-infection, simultaneously with a reduction in the levels of the ubiquitin-ligase HDM2. Taken together, these results suggest that CVB5 infection of HeLa cells elicit the intrinsic pathway of apoptosis by MDM2 degradation and p53 activation, destabilizing protein sumoylation, by a mechanism that is dependent on a functional ubiquitin-proteasome system.

2010-01-20

169

Novel microtubule-targeted agent 6-chloro-4-(methoxyphenyl) coumarin induces G2-M arrest and apoptosis in HeLa cells.  

UK PubMed Central (United Kingdom)

AIM: To identify a novel coumarin analogue with the highest anticancer activity and to further investigate its anticancer mechanisms. METHODS: The viability of cancer cells was investigated using the MTT assay. The cell cycle progression was evaluated using both flow cytometric and Western blotting analysis. Microtubule depolymerization was observed with immunocytochemistry in vivo and a tubulin depolymerization assay in vitro. Apoptosis was demonstrated using Annexin V/Propidium Iodide (PI) double-staining and sub-G(1) analysis. RESULTS: Among 36 analogues of coumarin, 6-chloro-4-(methoxyphenyl) coumarin showed the best anticancer activity (IC(50) value about 200 nmol/L) in HCT116 cells. The compound had a broad spectrum of anticancer activity against 9 cancer cell lines derived from colon cancer, breast cancer, liver cancer, cervical cancer, leukemia, epidermoid cancer with IC(50) value of 75 nmol/L-1.57 ?mol/L but with low cytotocitity against WI-38 human lung fibroblasts (IC(50) value of 12.128 ?mol/L). The compound (0.04-10 ?mol/L) induced G(2)-M phase arrest in HeLa cells in a dose-dependent manner, which was reversible after the compound was removed. The compound (10-300 ?mol/L) induced the depolymerization of purified porcine tubulin in vitro. Finally, the compound (0.04-2.5 ?mol/L) induced apoptosis of HeLa cells in dose- and time-dependent manners. CONCLUSION: 6-Chloro-4-(methoxyphenyl) coumarin is a novel microtubule-targeting agent that induces G(2)-M arrest and apoptosis in HeLa cells.

Ma YM; Zhou YB; Xie CM; Chen DM; Li J

2012-03-01

170

Representing life as opposed to being: the bio-objectification process of the HeLa cells and its relation to personalized medicine.  

UK PubMed Central (United Kingdom)

The immortal HeLa cells case is an intriguing example of bio-objectification processes with great scientific, social, and symbolic impacts. These cells generate questions about representation, significance, and value of the exceptional, variety, individuality, and property. Of frightening (a lethal cancer) and emarginated (a black, poor woman) origins, with their ability to "contaminate" cultures and to "spread" into spaces for becoming of extraordinary value for human knowledge, well-being, and economy advancements, HeLa cells have represented humanity, and emphasized the importance of individual as a core concept of the personalized medicine. Starting from the process leading from HeLa "cells" to HeLa "bio-objects," we focus on their importance as high quality bio-specimen. We discuss the tension between phenomenological characteristic of fundamental biological research and the variety of material and methodologies in epidemiology and personalized medicine. The emerging methodologies and societal changes reflect present EU policies and lead toward a new paradigm of science.

Svalastog AL; Martinelli L

2013-08-01

171

Surface glycosaminoglycans mediate adherence between HeLa cells and Lactobacillus salivarius Lv72.  

UK PubMed Central (United Kingdom)

BACKGROUND: The adhesion of lactobacilli to the vaginal surface is of paramount importance to develop their probiotic functions. For this reason, the role of HeLa cell surface proteoglycans in the attachment of Lactobacillus salivarius Lv72, a mutualistic strain of vaginal origin, was investigated. RESULTS: Incubation of cultures with a variety of glycosaminoglycans (chondroitin sulfate A and C, heparin and heparan sulfate) resulted in marked binding interference. However, no single glycosaminoglycan was able to completely abolish cell binding, the sum of all having an additive effect that suggests cooperation between them and recognition of specific adhesins on the bacterial surface. In contrast, chondroitin sulfate B enhanced cell to cell attachment, showing the relevance of the stereochemistry of the uronic acid and the sulfation pattern on binding. Elimination of the HeLa surface glycosaminoglycans with lyases also resulted in severe adherence impairment. Advantage was taken of the Lactobacillus-glycosaminoglycans interaction to identify an adhesin from the bacterial surface. This protein, identify as a soluble binding protein of an ABC transporter system (OppA) by MALDI-TOF/(MS), was overproduced in Escherichia coli, purified and shown to interfere with L. salivarius Lv72 adhesion to HeLa cells. CONCLUSIONS: These data suggest that glycosaminoglycans play a fundamental role in attachment of mutualistic bacteria to the epithelium that lines the cavities where the normal microbiota thrives, OppA being a bacterial adhesin involved in the process.

Martín R; Martín C; Escobedo S; Suárez JE; Quirós LM

2013-09-01

172

Corticotropin-releasing hormone (CRH) is expressed in the human cervical carcinoma cells (HeLa) and upregulates the expression of Fas ligand.  

UK PubMed Central (United Kingdom)

Corticotropin-releasing hormone acts as a stressor mediator in the human reproductive system. Corticotropin-releasing hormone (CRH) has been detected in several carcinomas of gynecological origin like breast, ovarian, and endometrial carcinomas. It was additionally shown that CRH could induce Fas ligand (FasL) expression in ovarian carcinoma cell lines. To determine whether CRH could also be expressed during cervical cancer development, we studied the expression of CRH using HeLa cells in an in vitro cervical cancer model. We further studied whether CRH could regulate FasL expression. In that context, HeLa cells were cultured in the presence or absence of 1 ?M CRH. CRH and FasL expressions were assessed by indirect immunofluorescence, reverse transcription PCR, and Western blot. The current results indicated that in HeLa cells, CRH can significantly induce both FasL transcription and FasL translation. Taking into account previous studies already establishing a connection between FasL expression and tumor immunoescape in cervical cancer, it can be concluded that such immunoescape could be CRH dependent.

Taliouri E; Vrekoussis T; Vergetaki A; Agorastos T; Makrigiannakis A

2013-02-01

173

Herpes simplex virus type 1 propagation in HeLa cells interrupted by Nelumbo nucifera.  

Science.gov (United States)

Inhibitory effects of ethanolic extracts from 10 Chinese herbs on herpes simplex virus type 1 (HSV-1) replication were investigated. By a bioassay-guided fractionation procedure, NN-B-5 was identified from seeds of N. nucifera. NN-B-5 significantly blocked HSV-1 multiplication in HeLa cells without apparent cytotoxicity. To elucidate the point in HSV-1 replication where arrest occurred, a set of key regulatory events leading to the viral multiplication was examined, including HSV-1 DNA synthesis and viral immediate early gene expressions. Data from polymerase chain reaction and Southern blotting showed that there were impairments of HSV-1 DNA replication in HeLa cells treated with NN-B-5. Results indicated that the production and mRNA transcription of infected cell protein (ICP) 0 and ICP4 were decreased in NN-B-5 treated HeLa cells. Results of an electrophoretic mobility shift assay demonstrated that NN-B-5 interrupted the formation of alpha-trans-induction factor/C1/Oct-1/GARAT multiprotein/DNA complexes. The mechanisms of antiviral action of NN-B-5 seem to be mediated, at least in part, through inhibition of immediate early transcripts, such as ICP0 and ICP4 mRNA and then blocking of all downstream viral products accumulation and progeny HSV-1 production. PMID:16132118

Kuo, Yuh-Chi; Lin, Yun-Lian; Liu, Chih-Peng; Tsai, Wei-Jern

2005-08-18

174

Automatic segmentation of HeLa cell images  

CERN Document Server

In this work, the possibilities for segmentation of cells from their background and each other in digital image were tested, combined and improoved. Lot of images with young, adult and mixture cells were able to prove the quality of described algorithms. Proper segmentation is one of the main task of image analysis and steps order differ from work to work, depending on input images. Reply for biologicaly given question was looking for in this work, including filtration, details emphasizing, segmentation and sphericity computing. Order of algorithms and way to searching for them was also described. Some questions and ideas for further work were mentioned in the conclusion part.

Urban, Jan

2011-01-01

175

Polyimidazole conjugated oligonucleotides reach the nucleus of HeLa cells.  

UK PubMed Central (United Kingdom)

Oligonucleotide models bearing 6, 12 or 18 histamine residues were synthesized on solid support and labeled with fluorescein. Only the oligo with 6 histamine residues showed a high uptake in HeLa cells with a nuclear localization. Experiment a 4 degrees C or with bafilomicyn A1 suggest that uptake proceeded by an endocytosis mechanism followed by a destabilization of the membrane. Once in the cytoplasm the oligo reached rapidly the nucleus.

Morvan F; Castex C; Vivès E; Imbach JL

2001-04-01

176

A ROS-mediated lysosomal-mitochondrial pathway is induced by a novel Amonafide analogue, 7c, in human Hela cervix carcinoma cells.  

UK PubMed Central (United Kingdom)

In this study, a novel naphthalimide derivative 7c was designed which is topo II inhibiting though owning weak DNA binders. It was shown that 7c could induce cancer cells apoptosis and have less cytotoxicity in normal human cell. Further investigations on Hela cells revealed that 7c could also induce ROS generation, lysosome rupture as well as cathepsin B release. Subsequent mitochondrial damages including mitochondrial membrane permeabilization and the release of cytochrome c were also found in 7c when treating with Hela cells. According to our data, 7c may act as a lead compound for potential anticancer drugs. The idea of naphthalimides modification may also provide a novel strategy for naphthalimides design.

Shen K; Sun L; Zhang H; Xu Y; Qian X; Lu Y; Li Q; Ni L; Liu J

2013-06-01

177

Effect of denture base resin extracts on HeLa cells growth in vitro  

Directory of Open Access Journals (Sweden)

Full Text Available Growth of HeLa cell culture in vitro was examined, in different concentrations of four resin materials extracts which are used for denture base making. Cell growth was evaluated through density, invert microscope counting, after 48 hours of incubation and through metabolic MTT test after 3 days. Extract was taken by incubation of material sample on 37 °C in physiological solution, for 72 hours. It is given weaker growth, reduction of adherent cells count and phenotypic changes of cells grown in presence of extracts from all examined materials. Extracts of examined materials increase number of phyllopodic extensions on dose dependent manner.

Kosti? Milena; Najman Stevo; Koci? Jelena S.; Kruni? Nebojša; Ajdukovi? Zorica; Petrovi? Dimitrije; An?elkovi? Maja

2008-01-01

178

The Effects of N-(4-hydroxyphenyl) Retinamide on Proliferation and Apoptosis of Hela Cells  

Directory of Open Access Journals (Sweden)

Full Text Available To investigate the effects of N-(4-hydroxyphenyl) retinamide (4HPR) on the proliferation and apoptosis of Hela cells, the cell growth was observed by SRB test, colony-forming test and nude mice carcinogenic test. Morphologic changes of apoptosis were observed under microscopes and the normal, apoptotic and necrotic cells were identified by fluorescence staining. The biochemical features and percentage of apoptosis were performed by flow cytometry (FCM) and DNA agarose gel electrophoresis. The results of SRB test, colony-forming test and nude mice carcinogenic test showed that 4HPR could inhibit the cells proliferation in vitro and in vivo (P

Xiao-Hong Han; Yan-Jun Xue; Shi-He Shao; Xian-Qian Li; Hua-Xi Xu

2011-01-01

179

Hela l-CaD is Implicated in the Migration of Endothelial Cells/Endothelial Progenitor Cells in Human Neoplasms  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Caldesmon (CaD) is a major actin-binding protein distributed in a variety of cell types. No functional differences among the isoforms in in vitro studies were found so far. In a previous study we found that the low molecular caldesmon isoform (Hela l-CaD) is expressed in endothelial cells (ECs)/endo...

Zheng, Ping-Pin; van der Weiden, Marcel; Kros, Johan M

180

N-acetylcysteine increases susceptibility of HeLa cells to bacterial invasion.  

UK PubMed Central (United Kingdom)

Serratia grimesii are non-pathogenic bacteria capable, however, to invade eukaryotic cells provided that they synthesize intracellular metalloprotease grimelysin (Bozhokina et al. [2011] Cell. Biol. Int. 35: 111-118). To elucidate how invasion of grimelysin containing bacteria depends on physiological state of host cells, we studied the effect of N-acetylcysteine (NAC) on susceptibility of HeLa cells to invasion by the wild-type S. grimesii and recombinant E. coli expressing grimelysin gene. Incubation of HeLa cells with 10 mM NAC resulted in changes of cell morphology and disassembly of actin cytoskeleton that were reversed when NAC was removed from the culture medium. Both in the presence of NAC and upon its removal, the entry of grimelysin producing bacteria increased by a factor of 1.5-2 and 3-3.5 for wild-type S. grimesii and recombinant E. coli, respectively. This effect does not correlate with cytoskeleton rearrangements but may be due to the NAC-induced up-regulation of cell surface receptors playing a role in cell adhesion and cell-cell junctions. A twofold difference in the efficiency of S. grimesii and recombinant E. coli to enter the NAC-treated cells suggests that the entry of the wild-type and recombinant bacteria occurs via different receptors which activity is differently affected by NAC.

Bozhokina E; Vakhromova E; Gamaley I; Khaitlina S

2013-07-01

 
 
 
 
181

Variability of the nucleoli number in the progenies of intact and UV-irradiated clonogenic Hela cells  

Energy Technology Data Exchange (ETDEWEB)

The coefficient of the ''nucleoli number'' character heritability (h/sup 2/) in the population of intact Hela cells equals 0.21 to 0.33. UV-irradiation enhances almost equally both the intraclonic and population variability of the nucleoli number and, as a result, the coefficient of the ''nucleoli number'' character heritability does not change in the population of UV-irradiated Hela cells.

Kucheryavaya, N.A.; Zavol' naya, E.S.; Vakhtin, Yu.B. (AN SSSR, Leningrad. Inst. Tsitologii)

1981-01-01

182

Variability of the nucleoli number in the progenies of intact and UV-irradiated clonogenic Hela cells  

International Nuclear Information System (INIS)

[en] The coefficient of the ''nucleoli number'' character heritability (h2) in the population of intact Hela cells equals 0.21 to 0.33. UV-irradiation enhances almost equally both the intraclonic and population variability of the nucleoli number and, as a result, the coefficient of the ''nucleoli number'' character heritability does not change in the population of UV-irradiated Hela cells

1981-01-01

183

Nucleotide sequences of two glutamine tRNAs from HeLa cells  

Energy Technology Data Exchange (ETDEWEB)

The authors reported previously that 4.5s RNA{sub H} is associated with poly A{sup +} RNAs of rodent cells. However, this molecule was not found in human, monkey, cat, mink, rabbit, or chicken cells. In HeLa cells, several 4S RNAs are released from the poly A{sup +} RNA fraction. They determined the nucleotide sequence of one of the most abundant 4s RNA species in this fraction. Nucleotide sequence analysis was performance. This RNA is 75 nucleotides long with 14 modified nucleotides. This sequence can be drawn in a clover-leaf structure typical of tRNA with the anticodon NUG.

Harada, Fumio; Matsubara, Minoru (Kanazawa Univ. (Japan)); Kato, Nobuyuki (National Cancer Center Research Institute, Tokyo (Japan))

1989-10-25

184

Anticancer Activity Test for Extracts of Sarang Semut Plant (Myrmecodya pendens) to HeLa and MCM-B2 Cells  

Directory of Open Access Journals (Sweden)

Full Text Available The aim of this study is to investigate anticancer activity of methanol extract (ethylacetate, n-buthanol and water partitions) and water extract from Sarang semut (local name), Myrmecodya pendens which is one of Rubiaceae family. Within Papua area (Indonesia), this medicinal plant has been used traditionally as alternative treatment for ulcer, tumor and cancer. In this study, the extracts of this plant were tested for their activities in some cancer cells (HeLa and MCM-B2 cell). The result showed that water extract of this plant has better anti cancer activity compared to other extracts. The IC50 value of water extract A is 27.61 ppm (HeLa) and 54.57 ppm (MCM-B2), while water extract B is 29.36 ppm (HeLa) and 74.20 ppm (MCM-B2). Our study concluded that polar extract (water) exhibited higher anticancer activity than non-polar extracts (ethylacetate and n-buthanol).

A. Soeksmanto; M.A. Subroto; H. Wijaya; P. Simanjuntak

2010-01-01

185

[Overexpression of hSav1 promotes Mst1-induced apoptosis in HeLa cells].  

UK PubMed Central (United Kingdom)

OBJECTIVE: To elucidate the effect of hSav1 expression on Mst1-mediated apoptosis in HeLa cells. METHODS: Plasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and cotransfected into HeLa cells. Triple immunofluorescent labeling of hSav1, Mst1 and nucleus was performed to determine their subcellular localization. Plasmids pCMV-HA-hSav1 and/or pcDNA/4TO-Flag-Mst1 were transfected into HeLa cells, and 36 hours later cisplatin (50 micromol/L) as a pro-apoptotic agent was added for 14 hours. Cell apoptosis was analyzed by annexin V/PI assay. RESULTS: Plasmids pCMV-HA-hSav1 and pcDNA/4TO-Flag-Mst1 were constructed and the authenticity of constructs was verified by sequencing. The binding in vitro showed that hSav1 could be detect from the anti-Mst1 immunoprecipitation complex. The immunofluorescent labeling showed that hSav1 and Mst1 had the same localization in cells. Overexpressed protein hSav1 did not induce a significant cell apoptosis. However, co-expression of hSav1 with Mst1 resulted in a significant increase of apoptosis above the level seen with Mst1 alone (24.5% +/- 2.4% vs. 39.3% +/- 4.0%, P < 0.05). CONCLUSION: Our findings indicate that hSav1 is a newly identified protein that interacts with Mst1 and augments Mst1-mediated apoptosis.

Li ZM; Liu WC; Dong S; Luo XL; Li XL; Tao DD; Gong JP; Hu JB

2009-07-01

186

Vibrio fluvialis attachs to but does not enter Hela cell monolayers  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english Considering the possibility that invasiveness could be a neglected factor of virulence in Vibrio fluvialis-linked enteritis, since a dysenteric form of the disease was seen in Bangladesh, we studied 12 Brazilian strains of the organism, six clinical and six environmental, to determine whether they might be able to enter into HeLa cell monolayers or would carry plasmids incidentally involved in invasiveness. Four human and two environmental isolates attached to but did not (more) enter into the cells. Though five strains harbored plasmids,no relationship was found between the carriage of these genetic elements and adhesiveness.

Carvalho, I. T.; Magalhães, V.; Leal, N. C.; Melo, V.; Magalhães, M.

1994-06-01

187

Vibrio fluvialis attachs to but does not enter Hela cell monolayers  

Directory of Open Access Journals (Sweden)

Full Text Available Considering the possibility that invasiveness could be a neglected factor of virulence in Vibrio fluvialis-linked enteritis, since a dysenteric form of the disease was seen in Bangladesh, we studied 12 Brazilian strains of the organism, six clinical and six environmental, to determine whether they might be able to enter into HeLa cell monolayers or would carry plasmids incidentally involved in invasiveness. Four human and two environmental isolates attached to but did not enter into the cells. Though five strains harbored plasmids,no relationship was found between the carriage of these genetic elements and adhesiveness.

I. T. Carvalho; V. Magalhães; N. C. Leal; V. Melo; M. Magalhães

1994-01-01

188

Glycosaminoglycans and syndecan-4 are involved in SDF-1/CXCL12-mediated invasion of human epitheloid carcinoma HeLa cells.  

UK PubMed Central (United Kingdom)

BACKGROUND: In addition to their physiologic effects in inflammation and angiogenesis, chemokines are involved in cancer pathology. The CXC-chemokine stromal cell-derived factor-1 (SDF-1)/CXCL12 mediates its biological activities through activation of G protein-coupled receptor CXCR4 and binds to glycosaminoglycans (GAGs). METHODS: Using Bio-coat cell migration chambers, specific antagonists, flow cytometry and RNA interference, we evaluate the involvement of heparan sulfate proteoglycans (HSPG) in the SDF-1/CXCL12-induced invasion of human cervix epitheloid carcinoma HeLa cells. RESULTS: The SDF-1/CXCL12-induced cell invasion is dependent on CXCR4. Furthermore, Protein Kinase C delta (PKC delta) and c-jun NH2-terminal kinase/stress-activated protein kinase (JNK/SAPK) are implicated in this event, but not extracellular signal-regulated kinase (ERK) 1/2. Moreover, the invasion of HeLa cells induced by SDF-1/CXCL12 was dependent on matrix metalloproteinase-9 (MMP-9). The pre-incubation of HeLa cells with heparin or with anti-heparan sulfate antibodies or with beta-d-xyloside inhibited SDF-1/CXCL12-mediated cell invasion. Furthermore, the down-regulation of syndecan-4, a heparan sulfate proteoglycan, decreased SDF-1/CXCL12-mediated HeLa cell invasion. GAGs, probably on syndecan-4, are involved in SDF-1/CXCL12-mediated cell chemotaxis. GENERAL SIGNIFICANCE: These data suggest that targeting the glycosaminoglycan/chemokine interaction could be a new therapeutic approach for carcinomas in which SDF-1/CXCL12 is involved.

Brule S; Friand V; Sutton A; Baleux F; Gattegno L; Charnaux N

2009-12-01

189

Tumor cell imaging using the intrinsic emission from PAMAM dendrimer: a case study with HeLa cells  

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HeLa 229 cells were treated with methotrexate (MTX) and doxorubicin (DOX), utilizing fourth generation (G4), amine terminated poly(amidoamine) {PAMAM} dendrimer as the drug carrier. In vitro kinetic studies of the release of both MTX and DOX in presence and absence of G4, amine terminated PAMAM dend...

Biswal, Bijesh K.; Kavitha, Manniledam; Verma, R. S.; Prasad, Edamana

190

Bordetella parapertussis invasion of HeLa 229 cells and human respiratory epithelial cells in primary culture.  

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Bordetella parapertussis, a respiratory tract pathogen commonly regarded as noninvasive, was found to invade HeLa 229 cell monolayers. Following treatment of the monolayers with gentamicin, numbers of viable B. parapertussis recovered were comparable to those of invasive Salmonella and Shigella isol...

Ewanowich, C A; Sherburne, R K; Man, S F; Peppler, M S

191

Proteomic changes induced by podophyllotoxin in human cervical carcinoma HeLa cells.  

Science.gov (United States)

Podophyllotoxin, a kind of lignan extracted from the Podophyllum plant, has been shown to inhibit the growth of various carcinoma cells. However, the molecular mechanism remains unclear. In this study, the inhibition of cell growth and changes in protein expression induced by podophyllotoxin were investigated in human cervical carcinoma HeLa cells. Our results demonstrate that Podophyllotoxin inhibits HeLa cell growth and induces apoptosis. By using proteomic techniques, seven proteins were found to be significantly regulated by podophyllotoxin compared to the untreated control; among them, four were down-regulated and three were up-regulated. All of the seven proteins were identified with peptide mass fingerprinting using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) after in-gel trypsin digestion. Five of these proteins are involved in protein metabolism, and the other two play roles in cell communication and signaling transduction pathways. It is suggested that the effect of podophyllotoxin on the growth of tumor cells is significantly related to the metabolism-associated proteins. PMID:23336514

Wang, Bochan; Chen, Lifeng; Zhen, Hong; Zhou, Li; Shi, Ping; Huang, Zhiwei

2013-01-01

192

Proteomic changes induced by podophyllotoxin in human cervical carcinoma HeLa cells.  

UK PubMed Central (United Kingdom)

Podophyllotoxin, a kind of lignan extracted from the Podophyllum plant, has been shown to inhibit the growth of various carcinoma cells. However, the molecular mechanism remains unclear. In this study, the inhibition of cell growth and changes in protein expression induced by podophyllotoxin were investigated in human cervical carcinoma HeLa cells. Our results demonstrate that Podophyllotoxin inhibits HeLa cell growth and induces apoptosis. By using proteomic techniques, seven proteins were found to be significantly regulated by podophyllotoxin compared to the untreated control; among them, four were down-regulated and three were up-regulated. All of the seven proteins were identified with peptide mass fingerprinting using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) after in-gel trypsin digestion. Five of these proteins are involved in protein metabolism, and the other two play roles in cell communication and signaling transduction pathways. It is suggested that the effect of podophyllotoxin on the growth of tumor cells is significantly related to the metabolism-associated proteins.

Wang B; Chen L; Zhen H; Zhou L; Shi P; Huang Z

2013-01-01

193

Interaction of radiation and AT 1727 in HeLa S-3 cells in culture  

Energy Technology Data Exchange (ETDEWEB)

The present cell culture studies were carried out to compare the cytotoxicity of AT 1727 and ICRF 159 in HeLa S-3 cells grown in monolayer and multicellular spheroid systems. The quantitative comparison of cell culture data demonstrated that AT 1727 was more cytotoxic than ICRF 159 at equimolar doses. The increased cytotoxicity at AT 1727 became apparent when the cell survival curves and growth rates of multicellular spheroids were compared at doses above 0.1 mM. When the spheriods were irradiated and exposed to AT 1727 (0.05 mM), there was a pronounced potentiation of radiation effects in the growth rate of spheroids. Similar treatment to ICRF 159 did not show any enhancement of radiation effects. There were also differential effects of AT 1727 and ICRF 159 on the cell cycle progression. At 1727 causes a G1 block as well as a G2 block in HeLa monolayers, while ICRF 159 only induces G2 block. These cell culture data may be useful for further in vivo tumor studies to determine the therapeutic index of combined radiation at AT 1727.

He, S.Q.; Kim, S.H.; Kim, J.H.

1985-06-01

194

Different behavior of protein B23/nucleophosmin and UBF in HeLa cells during apoptosis  

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Full Text Available The behavior of UBF (upstream binding factor) and nucleophosmin in HeLa and HeLa-Bcl-2 cells during apoptosis induced by TNF-?, emetine, and their mixture was investigated. A pronounced apoptosis was achieved only in HeLa cells treated with a mixture of the inducers. Immunoblotting analysis of UBF and nucleophosmin in samples containing different portions of cells with apoptotic nuclei was carried out. It showed that UBF was proteolytically cleaved giving a stable 76-kDa fragment. Increasing content of the fragment during apoptosis correlated with the level of cells containing apoptotic nuclei and with a decrease in the content of full-sized UBF. Determination of N- and C-terminal sequences of UBF and 76-kDa fragment allowed us not only to characterize UBF at the protein level, but also to describe the site of the apoptosis-specific proteolysis. Nucleophosmin did not undergo proteolytic cleavage during apoptosis and its content was unchanged even in a sample containing 100% of cells with apoptotic nuclei. However in cells reached terminal stages of apoptosis, the balance between mono- and oligomeric forms of nucleophosmin changed due to depletion of monomeric forms and appearance of two additional oligomeric forms with lower molecular weight.

Natalia M. Vladimirova; Natalia A. Potapenko

2011-01-01

195

Chronic infection of HeLa cells with Japanese encephalitis virus. General characteristics of the system.  

UK PubMed Central (United Kingdom)

Chronic infection of HeLa cells was induced by an attenuated variant of Japanese encephalitis (JE) virus (HeLa-K3 cell line). The chronic infection was characterized by alternating phases of degeneration and recovery of the cell monolayer. JE virus was regularly released into the medium of chronically infected cell cultures and virus-specific antigen was regularly demonstrated in the cytoplasm of 15--25% of cells. JE virus persisting in HeLa-K3 cells was sensitive to pancreatic ribonuclease and resistant to treatment with 4 M urea. HeLa-K3 cells did not undergo cytological or karyological transformation; they were susceptible to superinfection with heterologous viruses but resistant to reinfection with homologous virus.

Loginova NV; Deryabin PG; Mikhailova GR; Tsareva AA; Buinitskaya OB

1980-12-01

196

Chronic infection of HeLa cells with Japanese encephalitis virus. General characteristics of the system.  

Science.gov (United States)

Chronic infection of HeLa cells was induced by an attenuated variant of Japanese encephalitis (JE) virus (HeLa-K3 cell line). The chronic infection was characterized by alternating phases of degeneration and recovery of the cell monolayer. JE virus was regularly released into the medium of chronically infected cell cultures and virus-specific antigen was regularly demonstrated in the cytoplasm of 15--25% of cells. JE virus persisting in HeLa-K3 cells was sensitive to pancreatic ribonuclease and resistant to treatment with 4 M urea. HeLa-K3 cells did not undergo cytological or karyological transformation; they were susceptible to superinfection with heterologous viruses but resistant to reinfection with homologous virus. PMID:6111200

Loginova, N V; Deryabin, P G; Mikhailova, G R; Tsareva, A A; Buinitskaya, O B

1980-12-01

197

Nanomolar ouabain elicits apoptosis through a direct action on HeLa cell mitochondria.  

UK PubMed Central (United Kingdom)

The steroid Na(+)/K(+) ATPase (NKA) blocker ouabain has been shown to exhibit pro-apoptotic effects in various cell systems; however, the mechanism involved in those effects is unclear. Here, we have demonstrated that incubation of HeLa cells during 24h with nanomolar concentrations of ouabain or digoxin causes apoptotic death of 30-50% of the cells. Ouabain caused the activation of caspases-3/7 and -9; however, caspase-8 was unaffected. The fact that compound Z-LEHD-FMK reduced both apoptosis and caspase-9 activation elicited by ouabain, suggest a mitochondrially-mediated pathway. This was strengthened by the fact that ouabain caused ATP depletion and the release of mitochondrial cytochrome c into the cytosol. Furthermore, upon ouabain treatment mitochondrial disruption and redistribution into the cytosol were observed. A mitochondrial site of action for ouabain was further corroborated by tight co-localisation of fluorescent ouabain with mitochondria. Finally, in ouabain-treated cells the histamine-elicited elevation of cytosolic Ca(2+) concentration ([Ca(2+)]c) suggests an additional effect on the endoplasmic reticulum (ER) leading to Ca(2+) store depletion. We conclude that fluorescent ouabain is taken up and tightly co-localises with mitochondria of HeLa cells. This indicates that apoptosis may be triggered by a direct action of ouabain on mitochondria.

Alonso E; Cano-Abad MF; Moreno-Ortega AJ; Novalbos J; Milla J; García AG; Ruiz-Nuño A

2013-08-01

198

Nanomolar ouabain elicits apoptosis through a direct action on HeLa cell mitochondria.  

Science.gov (United States)

The steroid Na(+)/K(+) ATPase (NKA) blocker ouabain has been shown to exhibit pro-apoptotic effects in various cell systems; however, the mechanism involved in those effects is unclear. Here, we have demonstrated that incubation of HeLa cells during 24h with nanomolar concentrations of ouabain or digoxin causes apoptotic death of 30-50% of the cells. Ouabain caused the activation of caspases-3/7 and -9; however, caspase-8 was unaffected. The fact that compound Z-LEHD-FMK reduced both apoptosis and caspase-9 activation elicited by ouabain, suggest a mitochondrially-mediated pathway. This was strengthened by the fact that ouabain caused ATP depletion and the release of mitochondrial cytochrome c into the cytosol. Furthermore, upon ouabain treatment mitochondrial disruption and redistribution into the cytosol were observed. A mitochondrial site of action for ouabain was further corroborated by tight co-localisation of fluorescent ouabain with mitochondria. Finally, in ouabain-treated cells the histamine-elicited elevation of cytosolic Ca(2+) concentration ([Ca(2+)]c) suggests an additional effect on the endoplasmic reticulum (ER) leading to Ca(2+) store depletion. We conclude that fluorescent ouabain is taken up and tightly co-localises with mitochondria of HeLa cells. This indicates that apoptosis may be triggered by a direct action of ouabain on mitochondria. PMID:23933121

Alonso, Elba; Cano-Abad, María F; Moreno-Ortega, Ana J; Novalbos, Jesús; Milla, Juan; García, Antonio G; Ruiz-Nuño, Ana

2013-08-07

199

The effect of caffeine on x-ray repair of radioresistant HeLa cells  

International Nuclear Information System (INIS)

The contribution of caffeine-modifiable repair process to the radiosensitivity of a radioresistant HeLa strain (RC-355) has been investigated in comparison with control HeLa strain (CC-24). Both the final slope and the shoulder of X-ray survival curve for log-phase cells were affected by caffeine posttreatment. When the treatment with 10 mM caffeine delayed, an increase in survival was observed with increasing interval between irradiation and the treatment. During first several hours of the repair interval, the steepness of the final slope of survival curve decreased rapidly, and rate of the decrease was found to be higher in RC-355 than in CC-24 cells. Longer time (24 hours or more) before the initiation of caffeine treatment was required for the complete recovery of the shoulder. When the cells were incubated in plateau-phase after irradiation, an appreciable increase in survival was observed in comparison with when plated immediately following X-ray. The increase was found to be greater for RC-355 than for CC-24. The results suggest that the radioresistant RC-355 cells repaired more X-ray-induced PLD than CC-24 cells did. (author).

1985-01-01

200

Clonal heterogeneity in delayed decrease of plating efficiency of irradiated HeLa cells  

International Nuclear Information System (INIS)

[en] The clonogenic potential of progeny of irradiated HeLa cells was studied at different times after single doses of 4-12 Gy. The dose-dependent decrease in plating efficiency that was observed resembled the effect termed 'delayed lethal mutation' by Seymour et al. (1986). The effect decreased with time after irradiation. Individual clones of irradiated and non-irradiated cells were isolated, expanded and replated 5 weeks after irradiation, i.e., after between 200 000 and 1 000 000 progeny had formed from the individual parent cell. The plating efficiency of progeny of unirradiated cells did not vary much, whereas clonal progeny of irradiated cells had plating efficiencies ranging from 3% to 76%. The plating efficiency was not related to the cell number in the original clone. (orig.)

1993-01-01

 
 
 
 
201

Clonal heterogeneity in delayed decrease of plating efficiency of irradiated HeLa cells  

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The clonogenic potential of progeny of irradiated HeLa cells was studied at different times after single doses of 4-12 Gy. The dose-dependent decrease in plating efficiency that was observed resembled the effect termed 'delayed lethal mutation' by Seymour et al. (1986). The effect decreased with time after irradiation. Individual clones of irradiated and non-irradiated cells were isolated, expanded and replated 5 weeks after irradiation, i.e., after between 200 000 and 1 000 000 progeny had formed from the individual parent cell. The plating efficiency of progeny of unirradiated cells did not vary much, whereas clonal progeny of irradiated cells had plating efficiencies ranging from 3% to 76%. The plating efficiency was not related to the cell number in the original clone. (orig.).

Fitzek, M.; Trott, K.R. (Saint Bartholomew' s Medical Coll., London (United Kingdom). Dept. of Radiation Biology)

1993-01-01

202

Enhanced incorporation of radioactive inorganic phosphate into phospholipids of HeLa cells by tumor promoters  

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Teleocidin, a new tumor promoter, increased incorporation of radioactive inorganic phosphate (/sup 32/P/sub i/) into phospholipids in HeLa cells. This effect was detected within 1 h on incubation of the cells in medium containing teleocidin. The half-maximum effective dose of teleocidin was approximately 10 ng/ml. The main effect of teleocidin was on the incorporation of /sup 32/P/sub i/ into the phosphatidylcholine fraction, with a lesser effect on /sup 32/P/sub i/ incorporation into other phospholipid fractions. Increased incorporation of /sup 32/P/sub i/ into phospholipids was also observed on incubation of the cells with 12-O-tetradecanoylphorbol-13-acetate (TPA), dihydroteleocidin B, or lyngbyatoxin A, which are all complete tumor promoters, and also with mezerein, which is an incomplete and second stage promoter. On the other hand, at concentrations of up to 1 microgram/ml, 4-O-methyl TPA and C/sub a//sup 2/+ ionophore A23187, which are incomplete and first stage promoters, and phorbol, which has no promoting activity in skin carcinogenesis, did not cause any increased incorporation of /sup 32/P/sub i/ into phospholipid fractions of HeLa cells.

Nishino, H.; Fujiki, H.; Terada, M.; Sato, S.

1983-01-01

203

Role of NADPH oxidase in HeLa cell lesion induced by X-ray irradiation  

International Nuclear Information System (INIS)

[en] To investigate the role of NADPH oxidase in HeLa cell lesion induced by X-ray irradiation, the change of cell survival was detected with MTT assay, reactive oxygen species (ROS) was measured by flu- orospeetrophotometer. Immunostaining and confocal laser-scanning microscopy was employed to detect the co-localization of two subunit of NADPH oxidase, p47phox and gp91phox in the cell. Western blotting was used to detect the expression of gp91phox before and after X-ray irradiation. After X-ray irradiation, intracellular level of ROS increased obviously. But the increase could be blocked by diphenyleneiodonium (DPI), an inhibitor of NADPH oxidase. Meanwhile, cytosolic subunit p47phox moved to membrane and co-localizated with gp91phox after irradiation. Moreover, the results also show that gp91phox increased sharply after 12 Gy X-ray irradiation. Therefore, NADPH oxidase-mediated production of ROS plays an important role in HeLa cell lesion induced by X-ray. (authors)

2008-01-01

204

Biostimulation of HeLa cells by low-intensity visible light. Pt. 2  

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The action spectra of low-intensity light in the range from 300 to 900 nm on the synthesis rate of nucleic acids in the culture of HeLa cells has been measured. The synthesis of DNA and RNA is stimulated in several spectral intervals with maxima nearby 400, 630, 680, 760 and 820 nm. The stimulation effect is very sensitive to the irradiation duration (light intensity) at a fixed dose. The dose that causes the maximal stimulation is approximately 10 times smaller in the near-UV blue region than in red-IR region.

Karu, T.J. (AN SSSR,Troitsk. Laser Technology Center); Kalendo, G.S. (National Cancer Research Center, Moscow (USSR)); Letokhov, V.S.; Lobko, V.V. (AN SSSR, Troitsk. Inst. Spektroskopii)

1984-02-01

205

Increase of UV-resistance in xeroderma pigmentosum cells by human HeLaS3 DNA transfection  

International Nuclear Information System (INIS)

The DNA-mediated gene transfer had been carried out by both calcium phosphate coprecipitation and electroporation method. The cellular DNA and DNA fragments from human cervical carcinoma HeLaS3 cells were introduced with PSV2Neo DNA into XP20S (SV40) cells. The transfectants were picked up after twice selections by G418 and 3 J/m2 UV-irradiation. The results showed that cellular DNA and Bg1 I, Xho I digested DNA fragments from HeLaS3 could correct the deficiency of excision repair gene in XP cells, and cause the recipient cells resistant to UV irradiation. The second transfection experiment confirmed that HeLaS3 DNA were really integrated into XP cell chromosome and stably expressed within the cell genome.

1989-01-01

206

Quantum dots (QDs) restrain human cervical carcinoma HeLa cell proliferation through inhibition of the ROCK-c-Myc signaling.  

UK PubMed Central (United Kingdom)

Cancers often cause significant morbidity and even death to patients. To date, conventional therapies, such as chemotherapy, radiation and surgery, are often limited; meanwhile, novel anticancer therapeutics are urgently needed to improve clinical treatments. Rapid application of nanotechnology and nanomaterials represents a promising vista for the development of anti-cancer therapeutics. However, how to integrate the novel properties of nanotechnology and nanomaterials into cancer treatment warrants close investigation. In the current study, we report a novel finding about the inhibitory effect of CdSe quantum dots (QDs) on Rho-associated kinase (ROCK) activity in cervical carcinoma HeLa cells associated with the attenuation of the ROCK-c-Myc signaling. We mechanistically demonstrated that QD-conducted ROCK inhibition greatly diminished c-Myc protein stability due to reduced phosphorylation, and also suppressed its activity in transcribing target genes (e.g. HSPC111). Thus, the treatment of QDs greatly restrained HeLa cell growth by inducing cell cycle arrest at G1 phase due to the reduced ability of c-Myc in driving cell proliferation. Additionally, since HSPC111, one of the c-Myc targets, is involved in regulating cell growth through ribosomal biogenesis and assembly, the downregulation of HSPC111 could also contribute to diminished proliferation in HeLa cells upon QD treatment. These results together suggested that inhibition of ROCK activity or ROCK-mediated c-Myc signaling in tumor cells upon QD treatment might represent a promising strategy to restrain tumor progression for human cervical carcinoma.

Chen L; Qu G; Zhang C; Zhang S; He J; Sang N; Liu S

2013-03-01

207

The fibrate decreases radiation sensitivity via peroxisome proliferator-activated receptor ?-mediated superoxide dismutase induction in HeLa cells  

International Nuclear Information System (INIS)

The fibrates are ligands for peroxisome proliferator-activated receptor (PPAR) ? and used clinically as hypolipidemic drugs. The fibrates are known to cause peroxisome proliferation, enhance superoxide dismutase (SOD) expression and catalase activity. The antioxidant actions of the fibrates may modify radiation sensitivity. Here, we investigated the change of the radiation sensitivity in two cervix cancer cell lines in combination with fenofi brate (FF). Activity and protein expression of SOD were measured according to the concentration of FF. The mRNA expressions were measured by using real time reverse-transcription polymerase chain reaction. Combined cytotoxic effect of FF and radiation was measured by using clonogenic assay. In HeLa cells total SOD activity was increased with increasing FF doses up to 30 ?M. In the other hand, the catalase activity was increased a little. As with activity the protein expression of SOD1 and SOD2 was increased with increasing doses of FF. The mRNAs of SOD1, SOD2, PPAR? and PPAR? were increased with increasing doses of FF. The reactive oxygen species (ROS) produced by radiation was decreased by preincubation with FF. The surviving fractions (SF) by combining FF and radiation was higher than those of radiation alone. In Me180 cells SOD and catalase activity were not increased with FF. Also, the mRNAs of SOD1, SOD2, and PPAR? were not increased with FF. However, the mRNA of PPAR? was increased with FF. FF can reduce radiation sensitivity by ROS scavenging via SOD induction in HeLa. SOD induction by FF is related with PPAR?.

2012-01-01

208

Perturbation of enzymatic activity of the HeLa neoplastic cells by cytostatic active electromagnetic treatments  

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Full Text Available The study of interactions of low frequency and intensity electromagnetic fields (100 Hz and 5.5 mT of LFI- EMF) with some membrane bound and intracellular enzymatic biomolecules of HeLa cells has revealed an enhancement of membranary Na+-K+-ATP-ase, intracell LDH, SOD and ALP activities, as well as a repression of cellular ATP-ase, CAT, ACP activities in the case of cEMF treatment. Moreover, dcEMF has intensified the enzymatic activity of cellular ATP-ase, Px, CAT, has accentuated MDA levels and has also reduced the functioning degree of membranary Na+-K+- ATP-ase and of intracellular LDH, SOD and ALP. In the case of Px, GSH-Px and lipid peroxidation interference with both EMF variants, we assist to the induction of a stimulator effect upon their activities. The different sense and amplitude of reactivity of neoplastic cells enzymatic systems to the electromagnetic field irradiation were dependent on the EMF application mode (continuous or discontinuous). These variations of the enzymatic activities could be due to a direct or indirect interaction of exogenous cEMF or dcEMF with cellular (plasmalemma) or subcellular (organelles) structures and intracellular biomolecules (enzymes, DNA, RNA etc.), as well as a summation of the exogen and endogen electromagnetic fields effects. Thus, EMF induces a significant cytostatic effect either by alteration of the HeLa cells membranary or metabolic processes, or by intracellular increased generation of free radicals.

Cosmin Teodor Mihai; Daniela Gherghel; Gabriela Capraru; Elena Truta; Dorina Iurea; Ion Neacsu; Pincu Rotinberg

2010-01-01

209

Lycopodine from Lycopodium clavatum extract inhibits proliferation of HeLa cells through induction of apoptosis via caspase-3 activation.  

UK PubMed Central (United Kingdom)

Crude ethanolic extract of the plant Lycopodium clavatum has long been used in complementary and alternative medicine for treating various liver ailments and Alzheimer's disease. It has also been claimed to have potential anti-cancer properties in vivo in mice chronically fed liver carcinogens, p-dimethylamino azobenzene (initiator) and phenobarbital (promoter). Incidentally, crude ethanolic extract of Lycopodium clavatum is a mixture of some 201 alkaloids. In order to ascertain if any major fraction can be attributed to have pronounced anti-cancer effect, we examined this major fraction by eluting the crude extract in petroleum ether:ethyl aetate (17:3 vol/vol;) solvent and tried to understand its underlying mechanism. Studies on morphological changes, cell viability and cytotoxicity by microscopy and FACS, Western blot and immunofluorescence of Bcl-2, Bax, cytochrome c, caspase-3 were conducted. Lycopodine was found to induce chromatin condensation, inter-nucleosomal DNA fragmentation and enhanced cell population in sub-G1 region along with increase in reactive oxygen species generation and mitochondrial membrane potential depolarization, release of cytochrome c and activation of caspase-3 which are the events closely involved in apoptosis. An overall analysis of results showed that Lycopodine considerably inhibited growth of HeLa cells which indicates its potential use in chemotherapy.

Mandal SK; Biswas R; Bhattacharyya SS; Paul S; Dutta S; Pathak S; Khuda-Bukhsh AR

2010-01-01

210

Lycopodine from Lycopodium clavatum extract inhibits proliferation of HeLa cells through induction of apoptosis via caspase-3 activation.  

Science.gov (United States)

Crude ethanolic extract of the plant Lycopodium clavatum has long been used in complementary and alternative medicine for treating various liver ailments and Alzheimer's disease. It has also been claimed to have potential anti-cancer properties in vivo in mice chronically fed liver carcinogens, p-dimethylamino azobenzene (initiator) and phenobarbital (promoter). Incidentally, crude ethanolic extract of Lycopodium clavatum is a mixture of some 201 alkaloids. In order to ascertain if any major fraction can be attributed to have pronounced anti-cancer effect, we examined this major fraction by eluting the crude extract in petroleum ether:ethyl aetate (17:3 vol/vol;) solvent and tried to understand its underlying mechanism. Studies on morphological changes, cell viability and cytotoxicity by microscopy and FACS, Western blot and immunofluorescence of Bcl-2, Bax, cytochrome c, caspase-3 were conducted. Lycopodine was found to induce chromatin condensation, inter-nucleosomal DNA fragmentation and enhanced cell population in sub-G1 region along with increase in reactive oxygen species generation and mitochondrial membrane potential depolarization, release of cytochrome c and activation of caspase-3 which are the events closely involved in apoptosis. An overall analysis of results showed that Lycopodine considerably inhibited growth of HeLa cells which indicates its potential use in chemotherapy. PMID:19786013

Mandal, Sushil Kumar; Biswas, Raktim; Bhattacharyya, Soumya Sundar; Paul, Saili; Dutta, Suman; Pathak, Surajit; Khuda-Bukhsh, Anisur Rahman

2009-09-26

211

Kaempferol-7-O-beta-D-glucoside (KG) isolated from Smilax china L. rhizome induces G2/M phase arrest and apoptosis on HeLa cells in a p53-independent manner.  

Science.gov (United States)

Kaempferol-7-O-beta-D-glucoside (KG), a flavonoid glycoside, isolated from Smilax china L. rhizome, displayed marked anticancer activity on a panel of established cancer cells, of which, HeLa human cervix carcinoma cells were the most sensitive. Meanwhile, the cytotoxic effects of KG on normal human cells (HEK293 embryonic kidney cells and L-02 embryonic liver cells) were much smaller than on cancer cells. This work studied the molecular mechanisms underlying KG induced growth inhibition in HeLa cells. The results showed that KG induced G2/M phase growth arrest correlated with Cyclin B1 and Cdk1 decrease in a p53-independent manner, and also caused an increase in apoptosis, which was confirmed by characteristic morphological changes, evident DNA fragmentation, increased apoptotic sub-G1 population. Furthermore, inhibition of NF-kappaB nuclear translocation, up-regulation of Bax and down-regulation of Bcl-2, were observed in HeLa cells treated with KG, which indicated that the mitochondrial pathway was involved in the apoptosis signal pathway. In summary, KG displayed a significant anti-tumor effect through cell cycle arrest and apoptotic induction in HeLa cells, which suggested that KG might have therapeutic potential against cervix carcinoma. PMID:18343026

Xu, Wen; Liu, Jianwen; Li, Changlong; Wu, He-Zhen; Liu, Yan-Wen

2008-03-14

212

Hyperthermia enhances the reactivation of irradiated adenovirus in HeLa cells  

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The reactivation of U.V.-irradiated adenovirus 2 in HeLa cells is enhanced 8 to 9 fold if the cells are given a brief hyperthermic shock before infection. Maximum reactivation is achieved by heating for 10 min at 45.5 deg C and with a delay of 36 h between heating and infection. The induction process requires protein synthesis only during the 3 h period immediately following heating; cycloheximide does not prevent the expression of enhanced reactivation if added to the cells after this time. Heat-enhanced reactivation exhibits properties similar in some respects to radiation-enhanced reactivation and indicates an increased capacity of the heated cells to tolerate DNA damage.

Piperakis, S.M.; McLennan, A.G. (Liverpool Univ. (UK))

1984-02-01

213

Examination of HeLa cell contamination of human cell lines derived from primary hepatomas using glucose-6-phosphate dehydrogenase and lactate dehydrogenase isozymes.  

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Full Text Available Isozyme patterns of glucose-6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) in human cell lines derived from primary hepatomas were compared with those in HeLa cells. Some cell lines derived from primary hepatomas having type B G6PD showed one or two isozymes of LDH. On the other hand, HeLa cells having type A G6PD showed four LDH isozymes. These findings suggest that not only G6PD, but also LDH may be useful for the detection of HeLa cell contamination of a culture in some cases.

Tokiwa,Takayoshi; Kusaka,Yasunori; Muraoka,Atsushi; Sato,Jiro

1989-01-01

214

Construction of the plasmid for expression of ETA-EGFP fusion protein under control of the cytomegalovirus promoter and its effects in HeLa cells.  

Science.gov (United States)

Tumor-targeted vectors encoding toxic protein genes are promising tools for treating malignant tumors. We used the pEGFP-N1 vector to construct a novel plasmid (pCMV-ETA-EGFP) for eukaryotic expression of a truncated Pseudomonas aeruginosa exotoxin A (ETA) that is known to inhibit protein synthesis, and subsequently induce cell death, by inactivation of elongation factor-2. ETA was linked to the enhanced green fluorescent protein (EGFP) gene, and ETA-EGFP gene expression was driven by the cytomegalovirus (CMV) promoter. The time-lapse effects of pCMV-ETA-EGFP expression were examined in transiently transfected HeLa cells. HeLa cells transfected with pCMV-ETA-EGFP or cotransfected with pCMV-ETA-EGFP and small er, Cyrilliccapital IE, CyrillicGFP-N1 showed lower fluorescence intensity than cells transfected with pEGFP-N1 alone. Analysis of the number of dead cells further confirmed the highly toxic effect of the ETA-EGFP fusion protein on cells transfected with pCMV-ETA-EGFP or cotransfected with pCMV-ETA-EGFP and small er, Cyrilliccapital IE, CyrillicGFP-N1. ETA-EGFP fusion protein induced apoptotic cell death through the caspase-3 activation. By using the antibody against a marker nucleolar antigen A3 [Grigoryev, A.A., Bulycheva, T.I., Sheval, E.V., Kalinina, I.A., Zatsepina, O.V., 2008. Cytological indicators of the overall suppression of protein synthesis revealed by staining with new monoclonal antibody. Cell Tissue Biol. 2, 191-199], the distribution of which changes when HeLa cells are treated with known translation inhibitors, we obtained evidence to support the idea that protein synthesis is inhibited in transfected cells in situ. ETA-EGFP fusion protein was identified in lysates of transfected cells using anti-GFP-BL antibodies. Collectively, our results indicate that HeLa cells transfected with pCMV-ETA-EGFP synthesize the ETA-EGFP fusion protein that efficiently inhibits protein synthesis, leading to massive cell death by an apoptosis-mediated pathway with a participation of caspase-3. The constructed vector can be used in suicidal gene therapy of cancer and may also be useful for investigating the general effects of translational downregulation in human cancer cells. We also suggest a novel approach for detecting the activity of new vectors in transfected cells, which is based on the redistribution of nucleolar proteins in transfected cells. PMID:19527753

Glinka, Elena M; Andryushchenko, Alla S; Sapozhnikov, Alexander M; Zatsepina, Olga V

2009-06-13

215

Probing of cancer cell apoptosis by SERS and LSCM  

Science.gov (United States)

Surface enhanced Raman spectroscopy (SERS) can provide information of internal structures and chemical components from different kinds of samples. Laser scanning confocal microscopy (LSCM) can show morphologic information of samples by high-resolution optical images with different focal planes. In this paper, the dynamic variation of cancer cells (HELA cells) in the apoptosis was first studied by combining SERS and LSCM. After gold nanoparticles (GNPS) uptake, HELA cells were divided into two groups, and were respectively studied at six different time points of cell apoptosis period by SERS and LSCM. The LSCM images of HELA cells obtained at different time points were analyzed, and the morphology varieties of HELA cells apoptosis were obtained. It suggests that HELA cells apoptosis gradually in the apoptosis period until they died. In addition, Raman spectra of HELA cells measured at different time points were also compared. It shows that some Raman signal peaks shift, and FWHM of Raman peaks change too. The variation of internal structures and chemical constituents were analyzed according to the shifts and FWHM of the Raman peaks. The internal dynamic information and morphologic varieties from HELA cells apoptosis gained by combining SERS and LSCM will make us to understand cancer cell apoptosis throughly.

Kang, Jian; Gu, Huaimin

2009-07-01

216

Mechanistic aspects of fluorescent gold nanocluster internalization by live HeLa cells  

Science.gov (United States)

We have studied cellular uptake of ultrasmall fluorescent gold nanoclusters (AuNCs) by HeLa cells by confocal fluorescence microscopy in combination with quantitative image analysis. Water solubilized, lipoic acid-protected AuNCs, which had an overall hydrodynamic diameter of 3.3 nm and emitted fluorescence in the near-infrared region at ~700 nm, were observed to accumulate on the cell membrane prior to internalization. The internalization mechanisms were analyzed using inhibitors known to interfere with specific pathways. Cellular uptake of AuNCs is energy-dependent and involves multiple mechanisms: clathrin-mediated endocytosis and macropinocytosis appear to play a significant role, whereas the caveolin-mediated pathway contributes only to a lesser extent. Co-labeling of different cell organelles showed that intracellular trafficking of AuNCs mainly follows through endosomal pathways. The AuNCs were ultimately transferred to lysosomes; they were completely excluded from the nucleus even after 24 h.We have studied cellular uptake of ultrasmall fluorescent gold nanoclusters (AuNCs) by HeLa cells by confocal fluorescence microscopy in combination with quantitative image analysis. Water solubilized, lipoic acid-protected AuNCs, which had an overall hydrodynamic diameter of 3.3 nm and emitted fluorescence in the near-infrared region at ~700 nm, were observed to accumulate on the cell membrane prior to internalization. The internalization mechanisms were analyzed using inhibitors known to interfere with specific pathways. Cellular uptake of AuNCs is energy-dependent and involves multiple mechanisms: clathrin-mediated endocytosis and macropinocytosis appear to play a significant role, whereas the caveolin-mediated pathway contributes only to a lesser extent. Co-labeling of different cell organelles showed that intracellular trafficking of AuNCs mainly follows through endosomal pathways. The AuNCs were ultimately transferred to lysosomes; they were completely excluded from the nucleus even after 24 h. Electronic supplementary information (ESI) available: Effect of serum on the AuNC uptake by HeLa cells and colocalization result of AuNCs with the cell nucleus for 2-24 h. See DOI: 10.1039/c2nr33147k

Yang, Linxiao; Shang, Li; Nienhaus, G. Ulrich

2013-01-01

217

Apoptotic effect on HeLa Cells produced by Chlamydia trachomatis-LPS/ Efecto Apoptótico en Células HeLa Producido por el Lipopolisacárido (LPS) de Chlamydia trachomatis  

Scientific Electronic Library Online (English)

Full Text Available Abstract in spanish La interacción entre el lipopolisacárido (LPS) de Chlamydia trachomatis y las células de mamíferos permanece sin ser dilucidado. Chlamydia trachomatis es una bacteria intracelular responsable de diversas enfermedades en los humanos y animales. En este trabajo mediante el aislamiento del lipopolisacárido de dos serovares de Chlamydia trachomatis (LGV1-LGV2) y usando una coloración Supravital fluorescente (Hoechst 33258) fue posible investigar la respuesta de las cél (more) ulas HeLa. El efecto apoptótico que sufren este tipo de células fue visible cuando fueron expuestas a dicho LPS en concentraciones iguales o mayores que 0,5 µg/mL por un periodo de 48 horas, sin embargo se observó la falta de repuesta celular en su ausencia o en presencia de LPS de otras bacterias. Adicionalmente, el uso en iguales condiciones de polimyxina B conocido como un neutralizador de la acción del LPS demostró una disminución del efecto apoptótico en dichas células, indicando que la respuesta celular observada fue producida por C.trachomatis-LPS. Los resultados de este trabajo le dan fuerza a la teoría de que el LPS de C. trachomatis pudiera ser el responsable del efecto tóxico que se observa sobre las células cervicales infectadas con esta bacteria intracelular. Abstract in english The interaction between the lipopolysaccharide (LPS) of Chlamydia trachomatis and mammalian cells is still largely unknown. Chlamydia trachomatis is an obligate intracellular bacterium responsible for several diseases in humans and animals. In this work, thanks to the isolation of the lipopolysaccharide from two serovars of Chlamydia trachomatis (LGV1-LGV2) and using a nuclear supravital fluorescent stain (Hoechst 33258), it was possible to investigate the apoptotic effec (more) t on HeLa cells. This work shows the apoptotic effect on HeLa cells when they were exposed to C. trachomatis-LPS from two serovars at concentrations equal to or higher than 0.5 µg/mL for a period of 48h. and also the lack of cellular response in the absence of C. trachomatis-LPS or in the presence of LPS obtained from other bacteria. Additionally, the use in equal conditions of polymyxin B, known as an inhibitor of bacterial LPS, showed a decrease of the apoptotic effect in such cells indicating that the cellular response observed was produced by C. trachomatis-LPS. These results support the theory that the LPS from C. trachomatis could be responsible for the toxic effect on cervical cells infected by these bacteria.

Millán-Mendoza, Beatriz; Hakimi, Hamid; Eley, Adrian

2007-06-01

218

Chlamydia trachomatis Induces Remodeling of the Actin Cytoskeleton during Attachment and Entry into HeLa Cells  

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To elucidate the host cell machinery utilized by Chlamydia trachomatis to invade epithelial cells, we examined the role of the actin cytoskeleton in the internalization of chlamydial elementary bodies (EBs). Treatment of HeLa cells with cytochalasin D markedly inhibited the internalization of C. tra...

Carabeo, Reynaldo A.; Grieshaber, Scott S.; Fischer, Elizabeth; Hackstadt, Ted

219

Induction of unscheduled DNA synthesis in HeLa cells by allylic compounds.  

Science.gov (United States)

Thirteen allylic compounds, mostly with close structural relationship, were tested for their ability to induce unscheduled DNA synthesis (UDS) in HeLa cells and mutations in the Ames test; 11 induced UDS in dose dependence. Allyl isothiocyanate was negative in UDS (borderline in the Ames test) and acrolein (positive in the Ames test) proved toxic to HeLa cells, therefore UDS measurement was excluded. In general, positive qualitative and quantitative correlation between UDS, Ames test and alkylating properties (as measured in the 4-nitrobenzyl-pyridine test, NBP) were found. Among structural analogs and typical allylic compounds with various leaving groups, the amount of induced DNA repair at equimolar concentrations decreased in the same order as the mutagenic and alkylating activities in the other 2 test systems: 1,3-dichloropropene (cis) greater than 1,3-dichloropropene (trans) greater than 2,3-dichloro-1-propene; 1-chloro-2-butene greater than 3-chloro-1-butene greater than 3-chloro-2-methyl-1-propene greater than allyl chloride; allyl-methane-sulfonate greater than -iodide greater than -bromide greater than -chloride. PMID:6627227

Schiffmann, D; Eder, E; Neudecker, T; Henschler, D

1983-10-01

220

Bordetella parapertussis invasion of HeLa 229 cells and human respiratory epithelial cells in primary culture.  

UK PubMed Central (United Kingdom)

Bordetella parapertussis, a respiratory tract pathogen commonly regarded as noninvasive, was found to invade HeLa 229 cell monolayers. Following treatment of the monolayers with gentamicin, numbers of viable B. parapertussis recovered were comparable to those of invasive Salmonella and Shigella isolates. Invasion occurs through a cytochalasin-sensitive process which appears to be distinct from receptor-mediated endocytosis. Hyperimmune antisera raised against filaments hemagglutinin, a major adhesion of B. pertussis, did not inhibit invasion by B. parapertussis, suggesting that alternate adhesin(s) are required for invasion. In addition, B. parapertussis was found to invade human respiratory epithelial cells in primary culture, as demonstrated in ultrathin sections viewed by transmission electron microscopy. Although viable intracellular B. parapertussis persist within HeLa cells, they do not multiply there and the monolayers remain intact, suggesting a possible mechanism of carriage for these organisms.

Ewanowich CA; Sherburne RK; Man SF; Peppler MS

1989-04-01

 
 
 
 
221

Fatty acyl moieties: improving Pro-rich peptide uptake inside HeLa cells.  

UK PubMed Central (United Kingdom)

In the field of drug delivery there has been a continuous study of powerful delivery systems to aid non permeable drugs in reaching their intracellular target. Among the systems explored are cell penetrating peptides (CPPs), which first garnered interest a decade ago when the interesting translocation properties of the pioneer CPPs Tat and Antp were described. A new family of CPPs has recently been described as non cytotoxic Pro-rich vectors with favorable profiles for internalization in HeLa cells. Fatty acyl moieties that can tune a peptide's interaction with the lipophilic environment of a cell membrane have been incorporated into the Pro-rich sequence. Improvements in cellular uptake of peptides modified with fatty acyl groups, as studied by confocal microscopy and flow cytometry, as well as the results obtained by the interaction of these peptides with a model dioleoylphosphatidylcholine (DOPC) membrane and transmission electron microscopy (TEM), illustrate the importance of the fatty acyl moieties for efficient internalization.

Fernández-Carneado J; Kogan MJ; Van Mau N; Pujals S; López-Iglesias C; Heitz F; Giralt E

2005-06-01

222

Fatty acyl moieties: improving Pro-rich peptide uptake inside HeLa cells.  

Science.gov (United States)

In the field of drug delivery there has been a continuous study of powerful delivery systems to aid non permeable drugs in reaching their intracellular target. Among the systems explored are cell penetrating peptides (CPPs), which first garnered interest a decade ago when the interesting translocation properties of the pioneer CPPs Tat and Antp were described. A new family of CPPs has recently been described as non cytotoxic Pro-rich vectors with favorable profiles for internalization in HeLa cells. Fatty acyl moieties that can tune a peptide's interaction with the lipophilic environment of a cell membrane have been incorporated into the Pro-rich sequence. Improvements in cellular uptake of peptides modified with fatty acyl groups, as studied by confocal microscopy and flow cytometry, as well as the results obtained by the interaction of these peptides with a model dioleoylphosphatidylcholine (DOPC) membrane and transmission electron microscopy (TEM), illustrate the importance of the fatty acyl moieties for efficient internalization. PMID:15885117

Fernández-Carneado, J; Kogan, M J; Van Mau, N; Pujals, S; López-Iglesias, C; Heitz, F; Giralt, E

2005-06-01

223

Host cell responses to persistent mycoplasmas--different stages in infection of HeLa cells with Mycoplasma hominis.  

UK PubMed Central (United Kingdom)

Mycoplasma hominis is a facultative human pathogen primarily associated with bacterial vaginosis and pelvic inflammatory disease, but it is also able to spread to other sites, leading to arthritis or, in neonates, meningitis. With a minimal set of 537 annotated genes, M. hominis is the second smallest self-replicating mycoplasma and thus an ideal model organism for studying the effects of an infectious agent on its host more closely. M. hominis adherence, colonisation and invasion of HeLa cells were characterised in a time-course study using scanning electron microscopy, confocal microscopy and microarray-based analysis of the HeLa cell transcriptome. At 4 h post infection, cytoadherence of M. hominis to the HeLa cell surface was accompanied by differential regulation of 723 host genes (>2 fold change in expression). Genes associated with immune responses and signal transduction pathways were mainly affected and components involved in cell-cycle regulation, growth and death were highly upregulated. At 48 h post infection, when mycoplasma invasion started, 1588 host genes were differentially expressed and expression of genes for lysosome-specific proteins associated with bacterial lysis was detected. In a chronically infected HeLa cell line (2 weeks), the proportion of intracellular mycoplasmas reached a maximum of 10% and M. hominis-filled protrusions of the host cell membrane were seen by confocal microscopy, suggesting exocytotic dissemination. Of the 1972 regulated host genes, components of the ECM-receptor interaction pathway and phagosome-related integrins were markedly increased. The immune response was quite different to that at the beginning of infection, with a prominent induction of IL1B gene expression, affecting pathways of MAPK signalling, and genes connected with cytokine-cytokine interactions and apoptosis. These data show for the first time the complex, time-dependent reaction of the host directed at mycoplasmal clearance and the counter measures of this pestering pathogen.

Hopfe M; Deenen R; Degrandi D; Köhrer K; Henrich B

2013-01-01

224

Dihydroxy-isosteviol methyl ester from Pulsatilla nigricans induces apoptosis in HeLa cells: its cytoxicity and interaction with calf thymus DNA.  

UK PubMed Central (United Kingdom)

Dihydroxy-isosteviol methyl ester (DIME), the principal biological compound isolated from the medicinal plant Pulsatilla nigricans (Fam: Ranunculaceae) having the molecular formula of C21 H34 O3 (molecular weight 334.25), was administered to cervical cancer cells (HeLa) in vitro to evaluate its possible apoptotic (anti-cancer) potentials. We analyzed the expression of p53, Bax, Bcl2, Apaf and caspase 3 signal proteins and analyzed the early apoptotic events in HeLa cells induced by DIME using protocols like Annexin V-FITC and PI staining. DIME caused a significant decrease in cell viability, induced nuclear condensation and inter-nucleosomal DNA fragmentation. We further studied the interaction of DIME with calf thymus DNA as target through circular-dichroism spectra. Results showed that DIME interacted with DNA, bringing indiscernible changes in structure and conformation. Thus, DIME showed its capability to induce apoptosis in cancer cells, signifying its utility in drug design as a possible candidate for chemoprevention.

Das S; Das J; Samadder A; Khuda-Bukhsh AR

2013-05-01

225

Toxicity and Genotoxicity in HeLa and E. coli Cells Caused by a Helium Plasma Needle  

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Full Text Available The aim of this study is to determine the toxic and genotoxic damages produced by a helium plasma needle upon HeLa and E. coli (OG100 and PQ30) cell cultures. For HeLa cells survival (MTT) and microelectrophoresis comet) assays were performed; meanwhile in E. coli, viable count and genotoxicity by the chromotest were evaluated. The outcomes indicate that the plasma exposures on HeLa cells undergo more toxicity and genotoxicity as treatment time increases. With respect to E. coli, plasma exposure generated toxicity, but no genotoxicity could be detected with this system. In the strain OG100, defective in a protection mechanism to oxidizing agents, there was a reduction in the survival of one order of magnitude compared to the wild type strain PQ30. It suggests that such reduction is due to the plasma by means of the reactive oxygen and nitrogen species (RONS) generated during atmospheric air interaction.

E. Garcia-Alcantara; R. Lopez-Callejas; J. Serment-Guerrero; R. Pena-Eguiluz; A. E. Munoz-Castro; B. G. Rodriguez-Mendez; A. Mercado-Cabrera; R. Valencia-Alvarado; A. de-la-Piedad-Beneitez; J. M. E. Contreras-Ortiz; A. Barbabosa-Pliego

2013-01-01

226

Localization of non-native D-glyceraldehyde-3-phosphate dehydrogenase in growing and apoptotic HeLa cells.  

UK PubMed Central (United Kingdom)

Monoclonal antibodies that could not bind native tetramers of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) but could bind to dimeric, monomeric, or denatured forms of GAPDH were used to investigate its intracellular localization. These antibodies distinctly stained the nucleus in growing HeLa cells. In the cytoplasm, non-native GAPDH was colocalized with actin filaments. Incubation of HeLa cells with tumor necrosis factor ? (TNF-?) and the protein synthesis inhibitor emetine led to a drastic increase in the amount of the non-native GAPDH in the nuclei. Overproduction of Bcl-2 protein did not change the non-native GAPDH localization in the growing HeLa cells but prevented the development of apoptosis and the increase in the amount of non-native GAPDH in the nuclei upon incubation with TNF-?.

Arutyunova EI; Domnina LV; Chudinova AA; Makshakova ON; Arutyunov DY; Muronetz VI

2013-01-01

227

Suppression of postmitochondrial signaling and delayed response to UV-induced nuclear apoptosis in HeLa cells  

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Activation of postmitochondrial pathways by UV irradiation was examined using mouse lymphoma 3SB and human leukemic Jurkat cells and two human carcinoma cell lines (HeLa and MCF-7). Exposure of 3SB and Jurkat cells resulted in large amounts of cytochrome c and apoptosis-inducing factor (AIF) being released into the cytosol, and a clear laddering pattern of DNA fragments was observed within 3 h of incubation after irradiation. Simultaneously, activation of caspase-9 and its downstream caspases was detected. HeLa and MCF-7 cells also showed extensive release of mitochondrial factors and caspase-9 activation at 4 to 6 h after exposure, but apoptotic nuclear changes appeared much later. Compared with 3SB and Jurkat cells, these carcinoma cell lines exhibited reduced activation of caspase-9-like proteolytic activity by UV radiation, and levels of caspase-3-like activity in HeLa cells were extremely low, similar to those in caspase-3-deficient MCF-7 cells. These results suggest that the delayed response to UV-induced nuclear apoptosis in HeLa cells is due to a reduced activation of the caspase cascade downstream of cytochrome c release and suppression of caspase-3 activity. (author)

Sasai, Kaori; Yajima, Hirohiko; Suzuki, Fumio [Hiroshima Univ., (Japan). Research Inst. for Radiation Biology and Medicine

2002-03-01

228

Suppression of postmitochondrial signaling and delayed response to UV-induced nuclear apoptosis in HeLa cells  

International Nuclear Information System (INIS)

[en] Activation of postmitochondrial pathways by UV irradiation was examined using mouse lymphoma 3SB and human leukemic Jurkat cells and two human carcinoma cell lines (HeLa and MCF-7). Exposure of 3SB and Jurkat cells resulted in large amounts of cytochrome c and apoptosis-inducing factor (AIF) being released into the cytosol, and a clear laddering pattern of DNA fragments was observed within 3 h of incubation after irradiation. Simultaneously, activation of caspase-9 and its downstream caspases was detected. HeLa and MCF-7 cells also showed extensive release of mitochondrial factors and caspase-9 activation at 4 to 6 h after exposure, but apoptotic nuclear changes appeared much later. Compared with 3SB and Jurkat cells, these carcinoma cell lines exhibited reduced activation of caspase-9-like proteolytic activity by UV radiation, and levels of caspase-3-like activity in HeLa cells were extremely low, similar to those in caspase-3-deficient MCF-7 cells. These results suggest that the delayed response to UV-induced nuclear apoptosis in HeLa cells is due to a reduced activation of the caspase cascade downstream of cytochrome c release and suppression of caspase-3 activity. (author)

2002-01-01

229

Evaluation of hela cell lineage response to ? radiation from Holmium-166 embedded in ceramic seeds  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english This work studied the effects of ? radiation of Ho-166 embedded in ceramic seeds on HeLa cells. Methodology consisted in the production of ceramic seeds with holmium-165 by sol-gel route. Chemical and physical characterizations of the seeds were performed. Subsequently, nuclear characterization was performed by gamma spectrometry. Experimental and theoretical activities were defined and initial dose rate were evaluated by MIRD (Medical Internal Radiation Dose Committ (more) ee) methodology. The seeds were placed in confluent culture flasks and remained for six radionuclide half-lives. Biological results were represented by a clean 6 mm diameter area around the seed where the tumour cells were killed. The initial dose rate was 15.5 Gy. h-1. The maximum absorbed dose was 591.3 Gy. The features of the Ho-166 seeds suggested that such ceramic seeds were suitable for high dose rate brachytherapy.

Valente, Eduardo Sarmento; Cuperschmid, Ethel Mizrahy; Campos, Tarcisio Passos Ribeiro de

2011-10-01

230

The cytostatic potential confirmation of some fungal autochthonous biopreparations upon HeLa neoplastic cells cultures  

Directory of Open Access Journals (Sweden)

Full Text Available Some biopreparations of alkaloid-ergolinic nature have been extracted from hyphal and supernatant components, which were centrifugally separated from the submerged culture media of three strains of Claviceps purpurea (T1-3, T2-1 and T13-1), these being in different ontogenetic development stages (4, 6, 8, 10, and 12 days, respectively). In vitro testing of their interaction with cellular protein synthesis process of HeLa neoplastic cells cultures, has highlighted the cellular protein biosynthesis alteration, modifications of the protein dynamics sense and amplitude, as well as the cell cultures development inhibition. The protein synthesis inhibitory impact has confirmed the cytostatic action of these natural bioproducts, their cytostatic effectiveness being dependent of the Claviceps purpurea (T1-3, T2-1 and T13-1) strains specificity, the strain ontogenetic age, the biochemical nature of the intracellularly synthesized, stocked and extracellularly discharged substratum, as well as of their obtaining sources.

Pincu Rotinberg; Cosmin Teodor Mihai; Daniela Gherghel; Gabriela Capraru; Dorina Iurea; Craita Maria Rosu; Surdu Stefania

2010-01-01

231

Evaluation of hela cell lineage response to ? radiation from Holmium-166 embedded in ceramic seeds  

Directory of Open Access Journals (Sweden)

Full Text Available This work studied the effects of ? radiation of Ho-166 embedded in ceramic seeds on HeLa cells. Methodology consisted in the production of ceramic seeds with holmium-165 by sol-gel route. Chemical and physical characterizations of the seeds were performed. Subsequently, nuclear characterization was performed by gamma spectrometry. Experimental and theoretical activities were defined and initial dose rate were evaluated by MIRD (Medical Internal Radiation Dose Committee) methodology. The seeds were placed in confluent culture flasks and remained for six radionuclide half-lives. Biological results were represented by a clean 6 mm diameter area around the seed where the tumour cells were killed. The initial dose rate was 15.5 Gy. h-1. The maximum absorbed dose was 591.3 Gy. The features of the Ho-166 seeds suggested that such ceramic seeds were suitable for high dose rate brachytherapy.

Eduardo Sarmento Valente; Ethel Mizrahy Cuperschmid; Tarcisio Passos Ribeiro de Campos

2011-01-01

232

Translation In Vitro of Total Nuclear RNA from HeLa Cell Nuclei Infected with Adenovirus 2  

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Total heterogeneous nuclear RNA from HeLa cells infected with adenovirus for 18-20 hr stimulates amino acid incorporation into protein in a cell-free system from Ehrlich ascites tumors. This stimulation of protein synthesis by the nuclear RNA requires intact nuclei. The role of nuclei in this system...

Chatterjee, Nando K.; Weissbach, Herbert

233

Failure to detect "cap" structures in mitochondrial DNA-coded poly(A)-containing RNA from HeLa cells.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The structure of the 5'-termini has been investigated in mitochondrial DNA-coded poly(A)-containing RNA from HeLa cells. For this purpose, mitochondrial RNA isolated from cells labeled for 3 hours with [32P]orthophosphate in the presence of 20 microgram/ml camptothecin, and selected for poly(A) cont...

Grohmann, K; Amairic, F; Crews, S; Attardi, G

234

MicroRNA-433 negatively regulates the expression of thymidylate synthase (TYMS) responsible for 5-fluorouracil sensitivity in HeLa cells  

Science.gov (United States)

Background Thymidylate synthase (TYMS) is an important folate-dependent enzyme in DNA synthesis and an important target for cancer chemotherapy. High TYMS expression levels in tumors are generally associated with resistance to 5-fluorouracil (5-FU). The cause of the variability in TYMS expression is still not fully understood, however, only a small proportion of the TYMS expression can be explained by TYMS genetic polymorphisms. The purpose of this study is to identify novel microRNAs (miRNAs) which regulate the expression of TYMS and to determine whether miRNAs binding to the 3?-untranslated region (UTR) of TYMS mRNA affect the proliferation of HeLa cells treated with 5-FU. Methods An in silico search was performed to find potential binding sites of miRNAs in TYMS mRNA. The efficacy of predicted miRNAs at the 3?-UTR of TYMS mRNA was evaluated using a dual-luciferase reporter assay. TYMS mRNA and protein expression in HeLa cells was quantified with real-time RT-PCR and Western blotting, respectively. The effects of miR-433 on cell proliferative activity were determined by WST-8 assay. Results The overexpression of miR-433 was associated with significantly decreased reporter activity in the plasmid containing the 3?-UTR of TYMS mRNA (P?HeLa cells were significantly decreased by the overexpression of miR-433 (P?cell proliferation in HeLa cells treated with 5-FU at over 2.0 ?M. Conclusion The results indicate that miR-433 post-transcriptionally regulates the expression of TYMS mRNA and protein, and increases sensitivity to 5-FU in HeLa cells. This is the first report showing that a miRNA regulating TYMS expression has a significant impact on sensitivity to 5-FU treatment.

2013-01-01

235

MicroRNA-433 negatively regulates the expression of thymidylate synthase (TYMS) responsible for 5-fluorouracil sensitivity in HeLa cells.  

UK PubMed Central (United Kingdom)

BACKGROUND: Thymidylate synthase (TYMS) is an important folate-dependent enzyme in DNA synthesis and an important target for cancer chemotherapy. High TYMS expression levels in tumors are generally associated with resistance to 5-fluorouracil (5-FU). The cause of the variability in TYMS expression is still not fully understood, however, only a small proportion of the TYMS expression can be explained by TYMS genetic polymorphisms. The purpose of this study is to identify novel microRNAs (miRNAs) which regulate the expression of TYMS and to determine whether miRNAs binding to the 3[prime]-untranslated region (UTR) of TYMS mRNA affect the proliferation of HeLa cells treated with 5-FU. METHODS: An in silico search was performed to find potential binding sites of miRNAs in TYMS mRNA. The efficacy of predicted miRNAs at the 3[prime]-UTR of TYMS mRNA was evaluated using a dual-luciferase reporter assay. TYMS mRNA and protein expression in HeLa cells was quantified with real-time RT-PCR and Western blotting, respectively. The effects of miR-433 on cell proliferative activity were determined by WST-8 assay. RESULTS: The overexpression of miR-433 was associated with significantly decreased reporter activity in the plasmid containing the 3[prime]-UTR of TYMS mRNA (P < 0.01). The levels of TYMS mRNA and protein in HeLa cells were significantly decreased by the overexpression of miR-433 (P < 0.05). Furthermore, miR-433 increased inhibition of cell proliferation in HeLa cells treated with 5-FU at over 2.0 muM. CONCLUSION: The results indicate that miR-433 post-transcriptionally regulates the expression of TYMS mRNA and protein, and increases sensitivity to 5-FU in HeLa cells. This is the first report showing that a miRNA regulating TYMS expression has a significant impact on sensitivity to 5-FU treatment.

Gotanda K; Hirota T; Matsumoto N; Ieiri I

2013-08-01

236

Efficient induction of apoptosis in HeLa cells by a novel cationic porphycene photosensitizer.  

UK PubMed Central (United Kingdom)

In the present study we analyze the photobiological properties of 2,7,12-tris(?-pyridinio-p-tolyl)-17-(p-(methoxymethyl)phenyl) porphycene (Py3MeO-TBPo) in Hela cells, in order to assess its potential as a new photosensitizer for photodynamic therapy of cultured tumor cells. Using 0.5 ?M Py3MeO-TBPo, flow cytometry studies demonstrated an increase of intracellular drug levels related to the incubation time, reaching a maximum at 18 h. LysoTracker(®) Green (LTG) and MitoTracker(®) Green (MTG) probes were used to identify the subcellular localization. Upon exposure to ultraviolet excitation, red porphycene fluorescence was detected as red granules in the cytoplasm that colocalized with LTG. No significant toxic effects were detected for Py3MeO-TBPo in the dark at concentrations below 1 ?M. In contrast, Py3MeO-TBPo combined with red-light irradiation induced concentration- and fluence-dependent HeLa cells inactivation. Besides, all photodynamic protocols assayed induced a clear effect of cell detachment inhibition after trypsin treatment. Both apoptotic and necrotic cell death mechanisms can occur in HeLa cells depending on the experimental protocol. After 18 h incubation with 0.5 ?M Py3MeO-TBPo and subsequent red light irradiation (3.6 J/cm(2)), a high number of cells die by apoptosis, as evaluated by morphological alterations, immunofluorescent relocalization of Bax from cytosol to mitochondria, and TUNEL assay. Likewise, immunofluorescence techniques showed that cytochrome c is released from mitochondria into cytosol in cells undergoing apoptosis, which occurs immediately after relocation of Bax in mitochondria. The highest amount of apoptosis appeared 24 h after treatment (70%) and this cell death occurred without cell detachment to the substrate. In contrast, with 0.75 ?M Py3MeO-TBPo and 3.6 J/cm(2) irradiation, morphological changes showed a preferential necrotic cell death. Singlet oxygen was identified as the cytotoxic agent involved in cell photoinactivation. Moreover, cell cultures pre-exposed to the singlet oxygen scavenger sodium azide showed pronounced protection against the loss of viability induced by Py3MeO-TBPo and light. Different changes in distribution and organization of cytoskeletal elements (microtubules and actin microfilaments) as well as the protein vinculin, after apoptotic and necrotic photodynamic treatments have been analyzed. Neither of these two cell death mechanisms (apoptosis or necrosis) induced cell detachment. In summary, Py3MeO-TBPo appears to meet the requirements for further scrutiny as a very good photosensitizer for photodynamic therapy: it is water soluble, has a high absorption in the red spectral region (where light penetration in tissue is higher), and is able to induce effective high apoptotic rate (70%) related to the more widely studied photosensitizers.

Ruiz-González R; Acedo P; Sánchez-García D; Nonell S; Cañete M; Stockert JC; Villanueva A

2013-05-01

237

Efficient induction of apoptosis in HeLa cells by a novel cationic porphycene photosensitizer.  

Science.gov (United States)

In the present study we analyze the photobiological properties of 2,7,12-tris(?-pyridinio-p-tolyl)-17-(p-(methoxymethyl)phenyl) porphycene (Py3MeO-TBPo) in Hela cells, in order to assess its potential as a new photosensitizer for photodynamic therapy of cultured tumor cells. Using 0.5 ?M Py3MeO-TBPo, flow cytometry studies demonstrated an increase of intracellular drug levels related to the incubation time, reaching a maximum at 18 h. LysoTracker(®) Green (LTG) and MitoTracker(®) Green (MTG) probes were used to identify the subcellular localization. Upon exposure to ultraviolet excitation, red porphycene fluorescence was detected as red granules in the cytoplasm that colocalized with LTG. No significant toxic effects were detected for Py3MeO-TBPo in the dark at concentrations below 1 ?M. In contrast, Py3MeO-TBPo combined with red-light irradiation induced concentration- and fluence-dependent HeLa cells inactivation. Besides, all photodynamic protocols assayed induced a clear effect of cell detachment inhibition after trypsin treatment. Both apoptotic and necrotic cell death mechanisms can occur in HeLa cells depending on the experimental protocol. After 18 h incubation with 0.5 ?M Py3MeO-TBPo and subsequent red light irradiation (3.6 J/cm(2)), a high number of cells die by apoptosis, as evaluated by morphological alterations, immunofluorescent relocalization of Bax from cytosol to mitochondria, and TUNEL assay. Likewise, immunofluorescence techniques showed that cytochrome c is released from mitochondria into cytosol in cells undergoing apoptosis, which occurs immediately after relocation of Bax in mitochondria. The highest amount of apoptosis appeared 24 h after treatment (70%) and this cell death occurred without cell detachment to the substrate. In contrast, with 0.75 ?M Py3MeO-TBPo and 3.6 J/cm(2) irradiation, morphological changes showed a preferential necrotic cell death. Singlet oxygen was identified as the cytotoxic agent involved in cell photoinactivation. Moreover, cell cultures pre-exposed to the singlet oxygen scavenger sodium azide showed pronounced protection against the loss of viability induced by Py3MeO-TBPo and light. Different changes in distribution and organization of cytoskeletal elements (microtubules and actin microfilaments) as well as the protein vinculin, after apoptotic and necrotic photodynamic treatments have been analyzed. Neither of these two cell death mechanisms (apoptosis or necrosis) induced cell detachment. In summary, Py3MeO-TBPo appears to meet the requirements for further scrutiny as a very good photosensitizer for photodynamic therapy: it is water soluble, has a high absorption in the red spectral region (where light penetration in tissue is higher), and is able to induce effective high apoptotic rate (70%) related to the more widely studied photosensitizers. PMID:23517729

Ruiz-González, Rubén; Acedo, Pilar; Sánchez-García, David; Nonell, Santi; Cañete, Magdalena; Stockert, Juan Carlos; Villanueva, Angeles

2013-03-01

238

Differential gene expression in tumorigenic and nontumorigenic HeLa x normal human fibroblast hybrid cells.  

Science.gov (United States)

Fusion of tumorigenic HeLa cells with human skin fibroblasts results in chromosomally stable hybrids that are nontumorigenic and no longer express the HeLa tumor-associated marker intestinal alkaline phosphatase (IAP). Previous studies of spontaneous tumorigenic segregants from the nontumorigenic hybrids implicated the loss of one copy of human fibroblast chromosome 11 in the concomitant reexpression of tumorigenicity. In an attempt to identify genes involved in the control of tumorigenic expression, we performed differential display screening of nontumorigenic hybrid cells and tumorigenic segregants. Subsequent northern blot analyses reproducibly showed 17 differentially expressed genes, eight of which were expressed differentially in the nontumorigenic hybrids and nine of which were expressed differentially in the tumorigenic hybrids. The former were genes for 80K-L protein (a substrate of protein kinase C), AXL/UFO (a receptor tyrosine kinase), insulin-like growth factor binding protein 3, apolipoprotein AI regulatory protein, collagen type I alpha-2 chain, transforming growth factor-beta-induced gene product 3 (BIGH3), pregnancy-specific beta-1-glycoprotein, and fibroblast activation protein alpha. The latter nine genes were genes for serum/glucocorticoid-regulated kinase (SGK; a serine/threonine protein kinase), PTPCAAX1 (a tyrosine phosphatase), CXCR-4 (a G-protein-coupled membrane receptor), L-kynurenine hydrolase, beta-1, 4-galactosyltransferase, keratin 8, keratin 17, and H19 and a novel gene. The differential expression of these genes provided several interesting candidates for regulation of tumorigenic expression, including those involved in signal transduction and the extracellular matrix, cytoskeletal proteins, cell-surface enzyme, and the H19 gene. PMID:10569806

Tsujimoto, H; Nishizuka, S; Redpath, J L; Stanbridge, E J

1999-12-01

239

Apoptosis of HeLa and CaSki cell lines incubated with All-trans retinoid acid.  

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Full Text Available The aim of the study was to evaluate the concentrations of a soluble form of APO-1/Fas antigen (sFas, CD95) and a soluble Ligand for APO-1/Fas antigen (sCD95L, sFasL) in supernatants from CaSki and HeLa cell line cultures after the incubation with All-trans-retinoic acid. HPV-16 and HPV18 - positive cell lines were cultivated with All-trans-retinoic acid in concentrations of 1 x 10(-6) M/L and 1 x 10(-8) M/L. The cultures were incubated for 24 hours. Control culture with 3 microl of dimethyl-sulphoxide (DMSO) was incubated under identical conditions. The concentrations of soluble APO-1/Fas antigen and Fas Ligand in cell culture supernatants were estimated using immunoenzymatic methods. The obtained results showed significant decrease of concentrations of soluble APO-1/Fas antigen in supernatants from HeLa cell lines incubated with retinol in comparison with the control culture. Moreover, the concentrations of soluble Ligand for APO-1/Fas antigen in the supernatants of CaSki and HeLa cell lines were significantly lower in the culture incubated with All-trans retinoid acid when compared to the control culture. Higher concentrations of soluble APO-1/Fas antigen in supernatants from HeLa cell line without retinol may constitute a protective mechanism of the cells infected with the virus before undergoing Fas/FasL-dependent apoptosis. Lower concentrations of soluble APO-1/Fas antigen and soluble Ligand for APO-1/Fas in the supernatants from CaSki and HeLa cell cultures incubated with retinol suggest that retinoids can decrease the synthesis of soluble APO-1//Fas and soluble FasL in HPV-16 and HPV - 18 positive cells and that mechanisms protecting infected cells against Fas/FasL-mediated apoptosis become defective under the influence of retinol.

Dorota Darmochwal-Kolarz; Urszula Gasowska-Giszczak; Robert Paduch; Bogdan Kolarz; Piotr Wilci?ski; Jan Oleszczuk; Anna Kwasniewska

2010-01-01

240

Apoptosis of HeLa and CaSki cell lines incubated with All-trans retinoid acid.  

Science.gov (United States)

The aim of the study was to evaluate the concentrations of a soluble form of APO-1/Fas antigen (sFas, CD95) and a soluble Ligand for APO-1/Fas antigen (sCD95L, sFasL) in supernatants from CaSki and HeLa cell line cultures after the incubation with All-trans-retinoic acid. HPV-16 and HPV18 - positive cell lines were cultivated with All-trans-retinoic acid in concentrations of 1 x 10(-6) M/L and 1 x 10(-8) M/L. The cultures were incubated for 24 hours. Control culture with 3 microl of dimethyl-sulphoxide (DMSO) was incubated under identical conditions. The concentrations of soluble APO-1/Fas antigen and Fas Ligand in cell culture supernatants were estimated using immunoenzymatic methods. The obtained results showed significant decrease of concentrations of soluble APO-1/Fas antigen in supernatants from HeLa cell lines incubated with retinol in comparison with the control culture. Moreover, the concentrations of soluble Ligand for APO-1/Fas antigen in the supernatants of CaSki and HeLa cell lines were significantly lower in the culture incubated with All-trans retinoid acid when compared to the control culture. Higher concentrations of soluble APO-1/Fas antigen in supernatants from HeLa cell line without retinol may constitute a protective mechanism of the cells infected with the virus before undergoing Fas/FasL-dependent apoptosis. Lower concentrations of soluble APO-1/Fas antigen and soluble Ligand for APO-1/Fas in the supernatants from CaSki and HeLa cell cultures incubated with retinol suggest that retinoids can decrease the synthesis of soluble APO-1//Fas and soluble FasL in HPV-16 and HPV - 18 positive cells and that mechanisms protecting infected cells against Fas/FasL-mediated apoptosis become defective under the influence of retinol. PMID:20430726

Darmochwal-Kolarz, Dorota; Gasowska-Giszczak, Urszula; Paduch, Robert; Kolarz, Bogdan; Wilci?ski, Piotr; Oleszczuk, Jan; Kwasniewska, Anna

2009-01-01

 
 
 
 
241

Mitotic catastrophe induced in HeLa cells by photodynamic treatment with Zn(II)-phthalocyanine.  

Science.gov (United States)

Photodynamic therapy (PDT) is a tool against neoplastic and non-neoplastic diseases. PDT is capable to induce different cell death mechanisms in vitro, triggered in a dose-dependent manner. Relationships between PDT and apoptosis or necrosis induction are well-known, but other cell death mechanisms triggered after PDT are less understood. Here we present our results in p53-deficient human cervix carcinoma HeLa cells subjected to sublethal PDT treatments (mortality about 40%) using Zn(II)-phthalocyanine (ZnPc) incorporated into liposomes. We obtained a rapid metaphase blockage of cells that also showed clearly altered configurations of the mitotic spindle. Cell cycle arrest was followed by aneuploidisation and cell death with apoptotic morphology. Apoptosis was also confirmed by occurrence of PARP cleavage and Bax translocation to mitochondria. These features are components of the cell death mechanism known as mitotic catastrophe and represent, to our knowledge, the first description of this cell death modality after PDT with ZnPc. PMID:18497980

Rello-Varona, Santiago; Stockert, Juan Carlos; Cañete, Magdalena; Acedo, Pilar; Villanueva, Angeles

2008-06-01

242

Mitotic catastrophe induced in HeLa cells by photodynamic treatment with Zn(II)-phthalocyanine.  

UK PubMed Central (United Kingdom)

Photodynamic therapy (PDT) is a tool against neoplastic and non-neoplastic diseases. PDT is capable to induce different cell death mechanisms in vitro, triggered in a dose-dependent manner. Relationships between PDT and apoptosis or necrosis induction are well-known, but other cell death mechanisms triggered after PDT are less understood. Here we present our results in p53-deficient human cervix carcinoma HeLa cells subjected to sublethal PDT treatments (mortality about 40%) using Zn(II)-phthalocyanine (ZnPc) incorporated into liposomes. We obtained a rapid metaphase blockage of cells that also showed clearly altered configurations of the mitotic spindle. Cell cycle arrest was followed by aneuploidisation and cell death with apoptotic morphology. Apoptosis was also confirmed by occurrence of PARP cleavage and Bax translocation to mitochondria. These features are components of the cell death mechanism known as mitotic catastrophe and represent, to our knowledge, the first description of this cell death modality after PDT with ZnPc.

Rello-Varona S; Stockert JC; Cañete M; Acedo P; Villanueva A

2008-06-01

243

Overexpression of IGF-I receptor in HeLa cells enhances in vivo radioresponse  

International Nuclear Information System (INIS)

[en] Insulin-like growth factor I receptor (IGF-IR) is a transmembrane receptor tyrosine kinase whose activation strongly promotes cell growth and survival. We previously reported that IGF-IR activity confers intrinsic radioresistance in mouse embryo fibroblasts in vitro. However, it is still unclear whether tumor cells overexpressing IGF-IR exhibit radioresistance in vivo. For this purpose, we established HeLa cells that overexpress IGF-IR (HeLa-R), subcutaneously transplanted these cells into nude mice, and examined radioresponse in the resulting solid tumors. HeLa-R cells exhibited typical in vitro phenotypes generally observed in IGF-IR-overexpressing cells, as well as significant intrinsic radioresistance in vitro compared with parent cells. As expected, the transplanted HeLa-R tumors grew at a remarkably higher rate than parent tumors. Histological analysis revealed that HeLa-R tumors expressed more VEGF and had a higher density of tumor vessels. Unexpectedly, a marked growth delay was observed in HeLa-R tumors following 10 Gy of X-irradiation. Immunostaining of HeLa-R tumors for the hypoxia marker pimonidazole revealed a significantly lower level of hypoxic cells. Moreover, clamp hypoxia significantly increased radioresistance in HeLa-R tumors. Tumor microenvironments in vivo generated by the IGF-IR expression thus could be a major factor in determining the tumor radioresponse in vivo

2007-11-30

244

Terrein induces apoptosis in HeLa human cervical carcinoma cells through p53 and ERK regulation.  

Science.gov (United States)

Terrein, a fungal metabolite derived from Aspergillus terreus, has been shown to have a variety of biological activities in human cells including inhibition of melanogenesis, as well as anti-inflammatory, antioxidant and anticancer properties. In the present study, terrein was shown to have marked anticancer activity on HeLa human cervical carcinoma cells. Terrein exhibited inhibition of proliferation within the same ranges for other cancer cell types with an IC50 at 0.29 mM. The growth inhibition that induced cell death was via apoptosis mechanisms. Chromatin condensation was observed using the Hoechst 33342 stain, a DNA-specific dye. The increase of DNA fragmentation or the sub-G0 peak was also detected by flow cytometry. The signaling used by terrein to induce apoptosis was via the death-receptor and mitochondrial pathways; the cleavage of specific fluorogenic substrates by caspase-3, -8 and -9 activities are clearly demonstrated. The mitochondria were damaged as demonstrated by the decrease of the red/green ratio of the JC-1 staining and the increase of the Bax/Bcl-2 expression ratio. Further analysis of the upstream signaling by the quantitative real-time polymerase chain reaction showed that p53, p21 and ERK were upregulated which indicates the importance of their roles on terrein signaling. This study is the first to show that terrein has an effect on the anticancer properties in cervical cancer cells by inducing apoptosis through p53 and ERK regulation. Our data may help expand the function of the terrein compound and may also aid in the discovery of new anticancer agents. PMID:23417151

Porameesanaporn, Yuwarat; Uthaisang-Tanechpongtamb, Wanlaya; Jarintanan, Faongchat; Jongrungruangchok, Suchada; Thanomsub Wongsatayanon, Benjamas

2013-02-15

245

Terrein induces apoptosis in HeLa human cervical carcinoma cells through p53 and ERK regulation.  

UK PubMed Central (United Kingdom)

Terrein, a fungal metabolite derived from Aspergillus terreus, has been shown to have a variety of biological activities in human cells including inhibition of melanogenesis, as well as anti-inflammatory, antioxidant and anticancer properties. In the present study, terrein was shown to have marked anticancer activity on HeLa human cervical carcinoma cells. Terrein exhibited inhibition of proliferation within the same ranges for other cancer cell types with an IC50 at 0.29 mM. The growth inhibition that induced cell death was via apoptosis mechanisms. Chromatin condensation was observed using the Hoechst 33342 stain, a DNA-specific dye. The increase of DNA fragmentation or the sub-G0 peak was also detected by flow cytometry. The signaling used by terrein to induce apoptosis was via the death-receptor and mitochondrial pathways; the cleavage of specific fluorogenic substrates by caspase-3, -8 and -9 activities are clearly demonstrated. The mitochondria were damaged as demonstrated by the decrease of the red/green ratio of the JC-1 staining and the increase of the Bax/Bcl-2 expression ratio. Further analysis of the upstream signaling by the quantitative real-time polymerase chain reaction showed that p53, p21 and ERK were upregulated which indicates the importance of their roles on terrein signaling. This study is the first to show that terrein has an effect on the anticancer properties in cervical cancer cells by inducing apoptosis through p53 and ERK regulation. Our data may help expand the function of the terrein compound and may also aid in the discovery of new anticancer agents.

Porameesanaporn Y; Uthaisang-Tanechpongtamb W; Jarintanan F; Jongrungruangchok S; Thanomsub Wongsatayanon B

2013-04-01

246

Curing of HeLa cells persistently infected with equine arteritis virus by a peptide-conjugated morpholino oligomer.  

UK PubMed Central (United Kingdom)

A significant consequence of equine arteritis virus (EAV) infection of horses is persistence of the virus in a variable percentage of infected stallions. We recently established an in vitro model of EAV persistence in cell culture for the purpose of furthering our understanding of EAV biology in general and viral persistence in the stallion in particular. In this study we investigated whether persistently infected HeLa cells could be cured of EAV infection by treatment with an antisense peptide-conjugated phosphorodiamidate morpholino oligomer (PPMO) designed to target the 5'-terminal region of the EAV genome. We found that persistently infected HeLa cells passaged three times in the presence of 5-10 microM EAV-specific PPMO produced no detectable virus. The PPMO-cured HeLa cells were free of infectious virus, viral antigen and EAV RNA as measured by plaque assay, indirect immunofluorescence assay and RT-PCR, respectively. Furthermore, when re-challenged with EAV at several passages after discontinuation of PPMO treatments, PPMO-cured HeLa cells were found to be refractory to re-infection and to the re-establishment of viral persistence. While these findings demonstrate that PPMO can be used to eliminate persistent EAV infection in cell culture, the efficacy of PPMO against EAV in vivo remains to be addressed.

Zhang J; Stein DA; Timoney PJ; Balasuriya UB

2010-06-01

247

Curing of HeLa cells persistently infected with equine arteritis virus by a peptide-conjugated morpholino oligomer.  

Science.gov (United States)

A significant consequence of equine arteritis virus (EAV) infection of horses is persistence of the virus in a variable percentage of infected stallions. We recently established an in vitro model of EAV persistence in cell culture for the purpose of furthering our understanding of EAV biology in general and viral persistence in the stallion in particular. In this study we investigated whether persistently infected HeLa cells could be cured of EAV infection by treatment with an antisense peptide-conjugated phosphorodiamidate morpholino oligomer (PPMO) designed to target the 5'-terminal region of the EAV genome. We found that persistently infected HeLa cells passaged three times in the presence of 5-10 microM EAV-specific PPMO produced no detectable virus. The PPMO-cured HeLa cells were free of infectious virus, viral antigen and EAV RNA as measured by plaque assay, indirect immunofluorescence assay and RT-PCR, respectively. Furthermore, when re-challenged with EAV at several passages after discontinuation of PPMO treatments, PPMO-cured HeLa cells were found to be refractory to re-infection and to the re-establishment of viral persistence. While these findings demonstrate that PPMO can be used to eliminate persistent EAV infection in cell culture, the efficacy of PPMO against EAV in vivo remains to be addressed. PMID:20206215

Zhang, Jianqiang; Stein, David A; Timoney, Peter J; Balasuriya, Udeni B R

2010-03-03

248

Liquiritigenin induces mitochondria-mediated apoptosis via cytochrome c release and caspases activation in HeLa Cells.  

Science.gov (United States)

It has been demonstrated that many flavonoids possess a potent and broad spectrum of antitumor activity. Liquiritigenin is a flavanone extracted from Glycyrrhizae. This study investigated the effects of liquiritigenin on cell viability and apoptosis induction in human cervical carcinoma (HeLa) cells. The results show that liquiritigenin significantly suppressed cell proliferation in a dose- and time-dependent manner in HeLa cells. In addition, liquiritigenin promoted apoptosis in HeLa cells, evidenced by apoptotic morphological changes and Annexin-V binding. The apoptosis induction with liquiritigenin is associated with the up-regulation of p53 and Bax, along with down-regulation of Bcl-2 and survivin. Finally, examination of the mitochondrial pathway of apoptosis revealed that cytochrome c is released from mitochondria to cytosol, associated with the activation of caspase-9 and -3, and the cleavage of poly (ADP-ribose) polymerase (PARP). Overall, the results indicate that liquiritigenin induces apoptosis in part via the mitochondrial pathway, which is associated with p53 up-regulation, release of cytochrome c and elevated activity of caspase-9 and -3 in HeLa cells. PMID:20658471

Liu, Changwei; Wang, Yu; Xie, Sirou; Zhou, Yijing; Ren, Xiangmei; Li, Xiaoting; Cai, Yunqing

2011-02-01

249

Liquiritigenin induces mitochondria-mediated apoptosis via cytochrome c release and caspases activation in HeLa Cells.  

UK PubMed Central (United Kingdom)

It has been demonstrated that many flavonoids possess a potent and broad spectrum of antitumor activity. Liquiritigenin is a flavanone extracted from Glycyrrhizae. This study investigated the effects of liquiritigenin on cell viability and apoptosis induction in human cervical carcinoma (HeLa) cells. The results show that liquiritigenin significantly suppressed cell proliferation in a dose- and time-dependent manner in HeLa cells. In addition, liquiritigenin promoted apoptosis in HeLa cells, evidenced by apoptotic morphological changes and Annexin-V binding. The apoptosis induction with liquiritigenin is associated with the up-regulation of p53 and Bax, along with down-regulation of Bcl-2 and survivin. Finally, examination of the mitochondrial pathway of apoptosis revealed that cytochrome c is released from mitochondria to cytosol, associated with the activation of caspase-9 and -3, and the cleavage of poly (ADP-ribose) polymerase (PARP). Overall, the results indicate that liquiritigenin induces apoptosis in part via the mitochondrial pathway, which is associated with p53 up-regulation, release of cytochrome c and elevated activity of caspase-9 and -3 in HeLa cells.

Liu C; Wang Y; Xie S; Zhou Y; Ren X; Li X; Cai Y

2011-02-01

250

Analysis and prediction of pathways in HeLa cells by integrating biological levels of organization with systems-biology approaches.  

UK PubMed Central (United Kingdom)

It has recently begun to be considered that cancer is a systemic disease and that it must be studied at every level of complexity using many of the currently available approaches, including high-throughput technologies and bioinformatics. To achieve such understanding in cervical cancer, we collected information on gene, protein and phosphoprotein expression of the HeLa cell line and performed a comprehensive analysis of the different signaling pathways, transcription networks and metabolic events in which they participate. A total expression analysis by RNA-Seq of the HeLa cell line showed that 19,974 genes were transcribed. Of these, 3,360 were over-expressed, and 2,129 under-expressed when compared to the NHEK cell line. A protein-protein interaction network was derived from the over-expressed genes and used to identify central elements and, together with the analysis of over-represented transcription factor motifs, to predict active signaling and regulatory pathways. This was further validated by Metal-Oxide Affinity Chromatography (MOAC) and Tandem Mass Spectrometry (MS/MS) assays which retrieved phosphorylated proteins. The 14-3-3 family members emerge as important regulators in carcinogenesis and as possible clinical targets. We observed that the different over- and under-regulated pathways in cervical cancer could be interrelated through elements that participate in crosstalks, therefore belong to what we term "meta-pathways". Additionally, we highlighted the relations of each one of the differentially represented pathways to one or more of the ten hallmarks of cancer. These features could be maintained in many other types of cancer, regardless of mutations or genomic rearrangements, and favor their robustness, adaptations and the evasion of tissue control. Probably, this could explain why cancer cells are not eliminated by selective pressure and why therapy trials directed against molecular targets are not as effective as expected.

Higareda-Almaraz JC; Valtierra-Gutiérrez IA; Hernandez-Ortiz M; Contreras S; Hernandez E; Encarnacion S

2013-01-01

251

Analysis and Prediction of Pathways in HeLa Cells by Integrating Biological Levels of Organization with Systems-Biology Approaches  

Science.gov (United States)

It has recently begun to be considered that cancer is a systemic disease and that it must be studied at every level of complexity using many of the currently available approaches, including high-throughput technologies and bioinformatics. To achieve such understanding in cervical cancer, we collected information on gene, protein and phosphoprotein expression of the HeLa cell line and performed a comprehensive analysis of the different signaling pathways, transcription networks and metabolic events in which they participate. A total expression analysis by RNA-Seq of the HeLa cell line showed that 19,974 genes were transcribed. Of these, 3,360 were over-expressed, and 2,129 under-expressed when compared to the NHEK cell line. A protein-protein interaction network was derived from the over-expressed genes and used to identify central elements and, together with the analysis of over-represented transcription factor motifs, to predict active signaling and regulatory pathways. This was further validated by Metal-Oxide Affinity Chromatography (MOAC) and Tandem Mass Spectrometry (MS/MS) assays which retrieved phosphorylated proteins. The 14-3-3 family members emerge as important regulators in carcinogenesis and as possible clinical targets. We observed that the different over- and under-regulated pathways in cervical cancer could be interrelated through elements that participate in crosstalks, therefore belong to what we term “meta-pathways”. Additionally, we highlighted the relations of each one of the differentially represented pathways to one or more of the ten hallmarks of cancer. These features could be maintained in many other types of cancer, regardless of mutations or genomic rearrangements, and favor their robustness, adaptations and the evasion of tissue control. Probably, this could explain why cancer cells are not eliminated by selective pressure and why therapy trials directed against molecular targets are not as effective as expected.

Higareda-Almaraz, Juan Carlos; Valtierra-Gutierrez, Ilse A.; Hernandez-Ortiz, Magdalena; Contreras, Sandra; Hernandez, Erika; Encarnacion, Sergio

2013-01-01

252

Human papillomavirus type 16 and 18 L1 protein peptide binding to VERO and HeLa cells inhibits their VLPs binding.  

UK PubMed Central (United Kingdom)

Human papillomaviruses (HPVs) are the cause of epithelial lesions, HPV type 16 and type 18 being associated with the development of anogenital cancer. The L1 Major Capsid Protein (L1) represents about 90% of total HPV protein and is involved in virus-host cell interaction, but little is known about this binding process. L1 sequences from HPV types 16 and 18 were synthesized in 56 20-mer peptides, covering the entire protein, HPLC-purified, (125)I-radiolabeled and tested in VERO and HeLa cell-binding assays to identify those peptides with high specific binding activity. Peptides 18283 (residues 54-77) and 18294 (274-308) from HPV16 L1, as well as 18312 (59-78) and 18322 (259-278) from HPV18 L1, presented high specific target cell binding activity. Peptide 18283 and 18294 affinity constants were 300 and 600 nM, respectively. Enzyme cell treatment before binding assay indicated that VERO and HeLa cell peptide receptor is a surface-exposed protein. There was a 60% reduction in peptide 18283 binding to heparin lyase-treated cells. Cross-linking assays showed that these proteins molecular weights were around 69 and 54 kDa. Peptides 18283 and 18294 specifically inhibited HPV-16 VLP binding to HeLa cells. According to the L1- and VLP-reported structure, both peptides are close on the VLP-surface, belonging to the outer surface broad pockets suggested as being potential receptor sites. Furthermore, it has been reported that a conserved motif from peptide 18294 is the target for neutralizing antibodies. These results suggest that such binding sequences are used by the virus as cell-binding regions.

Vera-Bravo R; Ocampo M; Urquiza M; García JE; Rodríguez LE; Puentes A; López R; Curtidor H; Suárez JE; Torres E; Guzmán F; Díaz D; Cortes J; Bravo MM; Cómbita AL; Orozco O; Patarroyo ME

2003-11-01

253

Do altered activities of superoxide dismutases and the level of NF-kB modulate the effects of gamma radiation in HeLaS3 cells?  

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Full Text Available Most experimental models, including cell culture studies, have demon­strated that over-expression of manganese superoxide dismutase (MnSOD) in cells bearing a carcinoma phenotype has anti-proliferative and tumour suppression chara­cteristics. In contrast, when cervical carcinoma biopsies express MnSOD, there is a poor prognosis and resistance to radiation therapy. The results herein indicate that human cervical adenocarcinoma (HeLaS3) cells have increased MnSOD activity (up to 50 % of the total SOD activity) due to low expression of its repressor p53 and a high level of oxidative stress arising from the cell culture conditions. High MnSOD activity may be related to HeLaS3 cell radioresistance, illustrated by a high IC50 of 3.4 Gy and by a relatively high level of cell viability after gamma irradiation. In contrast to MnSOD activity, cytosolic CuZnSOD activity decreased after ionising radiation. The catalase (Cat) activity was unchanged. IR also increa­sed the nitric oxide synthase (NOS) activity. Such conditions lead to increased con­centrations of the superoxide radical, hydrogen peroxide and NO., which together may be responsible for the decreased expression of NF-kB and unaltered Cat ac­tivity. Therefore, the disturbed redox balance within HeLaS3 cells may be respon­sible for the cytotoxicity observed at higher irradiation doses. It could be concluded that inhibition of the CuZnSOD activity may be an important target for the selective killing of radioresistant cancer cells.

ANA NICIFOROVIC; MIROSLAV ADZIC; SNEZANA D. SPASI?; MARIJA B. RADOJCIC

2007-01-01

254

Uptake and quantification of intracellular concentration of lipophilic pro-oligonucleotides in HeLa cells.  

UK PubMed Central (United Kingdom)

Several lipophilic prodrugs of oligonucleotides (T12 and T20) bearing enzymolabile protecting groups and labeled with fluorescein were synthesized. Their cellular uptake was studied by three different approaches using fluorescence: fluorescence microscopy, flow cytometry and spectrofluorometry. The corresponding prooligonucleotides (pro-oligos) were rapidly and efficiently taken up by HeLa cells and were found homogeneously in the cytoplasm and in the nucleus. The uptake was proportional to their relative lipophilicity and likely proceeded through a passive diffusion mechanism. Uptake followed a dose-response curve. This prooligo approach led to a 2-log increase of uptake in comparison with a T20 phosphorothioate oligonucleotide. Finally, an intracellular concentration of pro-oligo was estimated between 4 and 6 microM for an external concentration of 1 microM and up to 27 microM for an incubation at 10 microM.

Bologna JC; Vivès E; Imbach JL; Morvan F

2002-02-01

255

THE LOCALIZATION BY ELECTRON MICROSCOPY OF HELA CELL SURFACE ENZYMES SPLITTING ADENOSINE TRIPHOSPHATE.  

UK PubMed Central (United Kingdom)

Cultures of normally proliferating Hela cells have been examined in thin sections by electron microscopy following glutaraldehyde fixation, staining in Wachstein and Meisel's adenosine triphosphate containing medium, postosmication, and embedding in an epoxy resin. The cells were stained in suspension in order to ensure uniform accessibility to reagents. Discrete localization of the enzyme reaction product (lead phosphate) was found at the plasma membranes of about half the cells, but nowhere else. It appeared in the form of an intensely electron-opaque deposit lying close against the outer surface of the cells and varying in amount from a chain of small particles to a dense band about 30 mmicro in width. This opaque reaction product was present over microvilli when absent elsewhere on a cell, was heaviest where microvilli and processes were profuse, and was minimal or lacking where cell surfaces were smooth. These observations are discussed in relation to both the idea that surface enzyme activity varies with the physiological phase of individual cells in a population, and the problem of how such enzyme activity becomes manifest at a given site on a morphologically changing membrane system.

EPSTEIN MA; HOLT SJ

1963-11-01

256

Defective caspase-3 activation and caspase-independent apoptosis in UV-irradiated HeLa S3 cells  

International Nuclear Information System (INIS)

Full text: Following exposure to radiation, most hematopoietic cells show a typical morphological characteristic of apoptosis and die quickly before the next mitosis. In contrast, death of non-hematopoietic cells, such as human tumor cells and fibroblasts, occurs after one or several cell divisions. Recently, it has been reported that many tumor cell lines die in interphase 12 h or longer after irradiation with relatively high doses of UV or ionizing radiation. However, the relationship between delayed interphase cell death and apoptosis is not clear. In this study, we used two kinds of cells, mouse lymphoma 3SB and human leukemic Jurkat cells and two human carcinoma cell lines (HeLa S3 and MCF-7). When irradiated with UV, 3SB and Jurkat cells showed an extensive release of cytochrome c from mitochondria and exhibited a clear production of oligonucleosomal DNA fragments within 3 h of incubation after irradiation. Simultaneously, activation of caspase-9 and its downstream caspases was detected. In the case of HeLa S3 and MCF-7 cells, DNA fragmentation could be detected at 24 or 48 h of post-irradiation incubation, but relatively early release of cytochrome c was observed (within 6 h after exposure). Interestingly, UV-irradiated HeLa S3 cells exhibited extremely low levels of caspase-3 like activity, similar to those in caspase-3-deficient MCF-7 cells, suggesting that the inhibition of apoptosis in HeLa S3 cells occurs downstream of cytochrome c release. A similar cell type-specific apoptosis was also observed when irradiated with ?-rays. To confirm the existence of caspase-independent apoptosis, we examined the effect of a caspase inhibitor, Z-VAD-FMK, on the induction of DNA fragmentation by UV exposure, and found that Z-VAD-FMK completely blocked DNA fragmentation in 3SB and Jurkat cells but did not suppress the delayed production of oligonuclesomal DNA fragments in HeLa S3 and MCF-7 cells. These data indicate that the delayed form of apoptosis in HeLa S3 cells occurs in a caspase-independent pathway

2003-01-01

257

Effects of Agaricus blazei Murill extract on sensitivity to chemotherapeutic agents in HeLa cells and its resistant sublines.  

Science.gov (United States)

Agaricus blazei Murill (ABM; Japanese name: Kawahiratake or Agarikusutake) extract is a widely used dietary supplement. However, limited information is available on the effects of the extract on the effectiveness of the chemotherapeutic agents. In this study, we examined the effects of ABM extract (Kyowa Wellness Co., Ltd.) on sensitivity to chemotherapeutic agents, paclitaxel and doxorubicin as MDR1/P-glycoprotein substrates, and cisplatin and 5-fluorouracil as non-substrates, in human cervical carcinoma HeLa cells, and paclitaxel-resistant and cisplatin-resistant derivatives (HeLa/TXL and HeLa/CDDP, respectively). The extract had no growth inhibitory effects on HeLa and the resistant cells at concentrations ranging from 7.6 × 10(-4) ? g/ml to 8.0 × 10(2)? g/ml, indicating no remarkable cytotoxic activity in vitro. In the presence of 0.1, 0.5, and 1 ? g/ml of ABM extract, sensitivity to paclitaxel, cisplatin and 5-fluorouracil did not change in HeLa, HeLa/TXL and HeLa/CDDP cells. However, the extract reduced sensitivity to doxorubicin in HeLa/TXL and HeLa/CDDP cells in a concentration-dependent manner. In conclusion, the concomitant use of ABM extract minimally affected sensitivity to various chemotherapeutic agents in HeLa cells and resistant sublines in vitro. PMID:22432463

Takara, Kohji; Shin, Yasunori; Obata, Yukihisa; Kitada, Noriaki; Ohnishi, Noriaki; Yokoyama, Teruyoshi

2008-01-01

258

Intracellular viscoelasticity of HeLa cells during cell division studied by video particle-tracking microrheology.  

Science.gov (United States)

Cell division plays an important role in regulating cell proliferation and differentiation. It is managed by a complex sequence of cytoskeleton alteration that induces dividing cells to change their morphology to facilitate their division. The change in cytoskeleton structure is expected to affect the intracellular viscoelasticity, which may also contribute to cellular dynamic deformation during cell division. However, the intracellular viscoelasticity during cell division is not yet well understood. In this study, we injected 100-nm (diameter) carboxylated polystyrene beads into the cytoplasm of HeLa cells and applied video particle tracking microrheology to measure their intracellular viscoelasticity at different phases during cell division. The Brownian motion of the intracellular nanoprobes was analyzed to compute the viscoelasticity of HeLa cells in terms of the elastic modulus and viscous modulus as a function of frequency. Our experimental results indicate that during the course of cell division, both intracellular elasticity and viscosity increase in the transition from the metaphase to the anaphase, plausibly due to the remodeling of cytoskeleton and redistributions of molecular motors, but remain approximately the same from the anaphase to the telophase. PMID:23864037

Chen, Yin-Quan; Kuo, Chia-Yu; Wei, Ming-Tzo; Wu, Kelly; Su, Pin-Tzu; Huang, Chien-Shiou; Chiou, Arthur

2014-01-01

259

Intracellular viscoelasticity of HeLa cells during cell division studied by video particle-tracking microrheology.  

UK PubMed Central (United Kingdom)

Cell division plays an important role in regulating cell proliferation and differentiation. It is managed by a complex sequence of cytoskeleton alteration that induces dividing cells to change their morphology to facilitate their division. The change in cytoskeleton structure is expected to affect the intracellular viscoelasticity, which may also contribute to cellular dynamic deformation during cell division. However, the intracellular viscoelasticity during cell division is not yet well understood. In this study, we injected 100-nm (diameter) carboxylated polystyrene beads into the cytoplasm of HeLa cells and applied video particle tracking microrheology to measure their intracellular viscoelasticity at different phases during cell division. The Brownian motion of the intracellular nanoprobes was analyzed to compute the viscoelasticity of HeLa cells in terms of the elastic modulus and viscous modulus as a function of frequency. Our experimental results indicate that during the course of cell division, both intracellular elasticity and viscosity increase in the transition from the metaphase to the anaphase, plausibly due to the remodeling of cytoskeleton and redistributions of molecular motors, but remain approximately the same from the anaphase to the telophase.

Chen YQ; Kuo CY; Wei MT; Wu K; Su PT; Huang CS; Chiou A

2014-01-01

260

Specific permeability and selective formation of gap junction channels in connexin-transfected HeLa cells  

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DNAs coding for seven murine connexins (Cx) (Cx26, Cx31, Cx32, Cx37, Cx40, Cx43, and Cx45) are functionally expressed in human HeLa cells that were deficient in gap junctional communication. We compare the permeabilities of gap junctions comprised of different connexins to iontophoretically injected...

 
 
 
 
261

Irradiation with He-Ne laser can influence the cytotoxic response of HeLa cells to ionizing radiation  

Energy Technology Data Exchange (ETDEWEB)

A monolayer of HeLa cells in plateau-phase of growth was exposed to He-Ne laser radiation (632.8 nm, 100 J/m[sup 2]) either 5, 60 or 180 min before [gamma]-irradiation (0.2-10 Gy, 6.75 Gy/min). (Author).

Karu, T.; Pyatibrat, L. (Academy of Sciences, Troitzk (Russian Federation). Laser Technology Center); Kalendo, G. (Russian Cancer Research Center, Moscow (Russian Federation))

1994-06-01

262

A heat-labile protein of Chlamydia trachomatis binds to HeLa cells and inhibits the adherence of chlamydiae.  

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From highly purified elementary bodies (EBs) of Chlamydia trachomatis, we have identified a protein of 38 kDa that selectively binds to monolayer cultures of HeLa cells. This protein, which we have named the chlamydial cytadhesin (CCA), is present on the surface of the EBs of three C. trachomatis se...

Joseph, T D; Bose, S K

263

Cellular uptake and biotransformation of arsenic V in transformed human cell lines HeLa S(3) and Hep G(2).  

UK PubMed Central (United Kingdom)

The biotransformation of arsenate was studied in two human transformed cell lines (epithelial HeLa S(3) and hepatoma Hep G(2) cells) by the determination of several arsenic species both in the culture medium and in the cells. Arsenate reduction, which was observed in the two cell lines, was higher in Hep G(2) cells. Methylation appeared to be a minor pathway for HeLa cells, but was more important in the hepatoma cells.

Peel AE; Brice A; Marzin D; Erb F

1991-01-01

264

Soluble ephrin a1 is necessary for the growth of HeLa and SK-BR3 cells  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Ephrin A1 (EFNA1) is a member of the A-type ephrin family of cell surface proteins that function as ligands for the A-type Eph receptor tyrosine kinase family. In malignancy, the precise role of EFNA1 and its preferred receptor, EPHA2, is controversial. Several studies have found that EFNA1 may suppress EPHA2-mediated oncogenesis, or enhance it, depending on cell type and context. However, little is known about the conditions that influence whether EFNA1 promotes or suppresses tumorigenicity. EFNA1 exists in a soluble form as well as a glycophosphatidylinositol (GPI) membrane attached form. We investigated whether the contradictory roles of EFNA1 in malignancy might in part be related to the existence of both soluble and membrane attached forms of EFNA1 and potential differences in the manner in which they interact with EPHA2. Results Using a RNAi strategy to reduce the expression of endogenous EFNA1 and EPHA2, we found that both EFNA1 and EPHA2 are required for growth of HeLa and SK-BR3 cells. The growth defects could be rescued by conditioned media from cells overexpressing soluble EFNA1. Interestingly, we found that overexpression of the membrane attached form of EFNA1 suppresses growth of HeLa cells in 3D but not 2D. Knockdown of endogenous EFNA1, or overexpression of full-length EFNA1, resulted in relocalization of EPHA2 from the cell surface to sites of cell-cell contact. Overexpression of soluble EFNA1 however resulted in more EPHA2 distributed on the cell surface, away from cell-cell contacts, and promoted the growth of HeLa cells. Conclusions We conclude that soluble EFNA1 is necessary for the transformation of HeLa and SK-BR3 cells and participates in the relocalization of EPHA2 away from sites of cell-cell contact during transformation.

Alford Spencer; Watson-Hurthig Adam; Scott Nadia; Carette Amanda; Lorimer Heather; Bazowski Jessa; Howard Perry L

2010-01-01

265

HeLa-S3 Cell Growth Conditions in Serum-Free Medium and Adaptability for Proliferation in Suspension Culture  

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Full Text Available Serum-free cell culture methods are now routinely support mammalian cell growth, a practice adopted for ethical, scientific and safety concerns. The HeLa-S3 cell line is a subclone of the HeLa cell line that can grow in Serum-Free Medium (SFM) as well as suspension cultures. In order to optimize its culturing conditions in SFM, the present study investigated the efficacy of insulin and L-glutamine additives as biotic factors as well as osmotic stress as abiotic factor, all affecting growth kinetics and metabolism. Insulin was used with different concentrations ranging from 10 to 50 mg L-1. It was found that cell growth is dependent on insulin up to a concentration of 25 mg L-1 at which maximum cell number as well as cell viability were achieved. Similarly, L-glutamine was used in the range of 3 to 8 mmol L-1 and was found optimum at 3 mmol L-1. However, osmotic stress using saline solution addition showed that osmolality in the range of 314 to 350 mOsm kg-1 is preferable to cells. The study also showed the successful sequential cell adaptation from adherent culture mode to suspension culture in which cells were able to grow in small clumps of spherical-shaped cells. Based on this cultivation strategy, HeLa-S3 cells were completely adapted to proliferate suspended in serum-free medium with sustained growth kinetics and physiological properties.

Sherif H. Abdeen; Ahmed M. Abdeen; Hesham A. El-Enshasy; Abdalla A. El Shereef

2011-01-01

266

C60-ToF SIMS imaging of frozen hydrated HeLa cells.  

UK PubMed Central (United Kingdom)

Sample preparation continues to be a major challenge for secondary ion mass spectrometry studies of biological materials. Maintaining the native hydrated state of the material is important for preserving both chemical and spatial information. Here, we discuss a method which combines a sample wash and dry protocol discussed by Berman et al (1 (1)) followed by plunge freezing in liquid ethane for a frozen-hydrated analysis of mammalian cells (HeLa). This method allows for the removal of the growth media and maintains the hydrated state of the cells so that they can be prepared frozen-hydrated without the need for a freeze-fracture device. The cells, which were grown on silicon, have been successfully re-grown after the cleaning procedure, confirming that a significant portion of the cells remain undamaged during the wash and dry. Results from preliminary SIMS measurements show that is it possible to detect a large variety of bio-molecular signals, including intact lipids from the plasma membrane in the mass range of 700-900 Da from single cells, with little external water interference at the surface.

Piwowar AM; Keskin S; Delgado MO; Shen K; Hue JJ; Lanekoff I; Ewing AG; Winograd N

2013-01-01

267

Effects of hyperthermia on the sedimentation of nucleoids from HeLa cells in sucrose gradients  

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The changes in sedimentation distance of nucleoids in neutral sucrose gradients in the presence of various concentrations of the intercalating dye ethidium bromide are believed to reflect changes in the amount of DNA supercoiling within the nucleoid. When HeLa cells were heated to 45/sup o/C for 30 min before cell lysis, sedimentation of their nucleoids was markedly affected; at all concentrations of ethidium bromide in the gradient, nucleoids from heated cells sedimented farther than those from unheated cells. After cellular protein was labeled with (/sup 3/H)leucine for 15 to 17 hr, approximately 4% of the total /sup 3/H radioactivity cosedimented with nucleoids from unheated cells. This percentage increased with heating time (10-70 min) and temperature (44-46/sup o/C); e.g., after 20 min at 45/sup o/C the fraction of /sup 3/H cosedimenting with nucleoids was 7%. The heat-induced change in nucleoid sedimentation distance was a linear function of the percentage of /sup 3/H cosedimenting with nucleoids. These results show that hyperthermia causes an increase in the protein content of nucleoids. The additional protein affects sedimentation by increasing the mass of the nucleoids and by decreasing the amount of DNA available for supercoiling and thereby the efficiency of DNA rewinding.

Roti Roti, J.L. (Univ. of California, San Francisco); Painter, R.B.

1982-01-01

268

DNA synthesis in generation 1 in x-irradiated HeLa cells  

International Nuclear Information System (INIS)

Measurements of DNA replication in a line of HeLa S3 cells during the generation (Generation 1) following that in which the cells are irradiated with 500 rad of 220-kV x rays (Generation 0) were carried out according to a number of different experimental protocols. These involved preirradiation labeling of the cells with low levels of [14C] thymidine in Generation-1 to provide a measure of the template DNA, synchronization by mitotic collection in Generation 0, resynchronization by either mitotic recollection or temporary drug-induced blockages in Generation 1, and either labeled-thymidine incorporation or density-label transfer during Generation 1. The results show that those cells that progress through S phase of Generation 1 and divide at the next mitosis replicate a full complement of DNA. However, apparent deficits of as much as 45% are measured if resynchronization in Generation 1 is effected by drug tretment following manipulations of the culture in the presence of particular media and drugs during Generation 0. These are attributed to combined radiation- and drug-induced disturbances in cell progression

1981-01-01

269

Cytochrome C-dependent Fas-independent apoptotic pathway in HeLa cells induced by delta12-prostaglandin J2.  

UK PubMed Central (United Kingdom)

Cyclopentenone prostaglandins (PGs) have antiproliferative activity on various tumor cell growth in vitro. Particularly, 9-deoxy-delta(9,12)-13,14-dihydro PGD(2) (delta(12)-PGJ(2)) was reported for its antineoplastic and apoptotic effects on various cancer cells, but its mechanism inducing apoptosis is still not clear. In this study, we have characterized apoptosis induced by delta(12)-PGJ(2) in HeLa cells. Treatment of delta(12)-PGJ(2) induced apoptosis as indicated by DNA fragmentation, chromatin condensation, and formation of apoptotic body. We also observed release of cytochrome c from mitochondria and activation of caspase cascade including caspase-3, -8, and -9. And the pan-caspase inhibitor z-Val-Ala-Asp (OMe) fluoromethyl-ketone (z-VAD-fmk) and Q-Val-Asp (OMe)-CH(2)-OPH (Q-VD (OMe)-OPH) prevented cell death induced by delta(12)-PGJ(2) showing participation of caspases in this process. However, protein expression level of Bcl-2 family was not altered by delta(12)-PGJ(2), seems to have no effect on HeLa cell apoptosis. And ZB4, an antagonistic Fas-antibody, exerted no effect on the activation of caspase 8 indicating that Fas receptor-ligand interaction was not involved in this pathway. Treatment of delta(12)-PGJ(2) also leads to suppression of nuclear factor kappaB (NF-kappaB) as indicated by nuclear translocation of p65/RelA and c-Rel and its DNA binding ability analyzed by EMSA. Taken together, our results suggest that delta(12)-PGJ(2)-induced apoptosis in HeLa cell utilized caspase cascade without Fas receptor-ligand interaction and accompanied with NF-kappaB inactivation.

Kim BE; Roh SR; Kim JW; Jeong SW; Kim IK

2003-08-01

270

Cytochrome C-dependent Fas-independent apoptotic pathway in HeLa cells induced by delta12-prostaglandin J2.  

Science.gov (United States)

Cyclopentenone prostaglandins (PGs) have antiproliferative activity on various tumor cell growth in vitro. Particularly, 9-deoxy-delta(9,12)-13,14-dihydro PGD(2) (delta(12)-PGJ(2)) was reported for its antineoplastic and apoptotic effects on various cancer cells, but its mechanism inducing apoptosis is still not clear. In this study, we have characterized apoptosis induced by delta(12)-PGJ(2) in HeLa cells. Treatment of delta(12)-PGJ(2) induced apoptosis as indicated by DNA fragmentation, chromatin condensation, and formation of apoptotic body. We also observed release of cytochrome c from mitochondria and activation of caspase cascade including caspase-3, -8, and -9. And the pan-caspase inhibitor z-Val-Ala-Asp (OMe) fluoromethyl-ketone (z-VAD-fmk) and Q-Val-Asp (OMe)-CH(2)-OPH (Q-VD (OMe)-OPH) prevented cell death induced by delta(12)-PGJ(2) showing participation of caspases in this process. However, protein expression level of Bcl-2 family was not altered by delta(12)-PGJ(2), seems to have no effect on HeLa cell apoptosis. And ZB4, an antagonistic Fas-antibody, exerted no effect on the activation of caspase 8 indicating that Fas receptor-ligand interaction was not involved in this pathway. Treatment of delta(12)-PGJ(2) also leads to suppression of nuclear factor kappaB (NF-kappaB) as indicated by nuclear translocation of p65/RelA and c-Rel and its DNA binding ability analyzed by EMSA. Taken together, our results suggest that delta(12)-PGJ(2)-induced apoptosis in HeLa cell utilized caspase cascade without Fas receptor-ligand interaction and accompanied with NF-kappaB inactivation. PMID:14508070

Kim, Bo-Eun; Roh, Sung Rae; Kim, Jin Woo; Jeong, Seong-Whan; Kim, In-Kyung

2003-08-31

271

The epimer of kaurenoic acid from Croton antisyphiliticus is cytotoxic toward B-16 and HeLa tumor cells through apoptosis induction.  

Science.gov (United States)

Cancer has become the leading cause of death in developing countries due to increased life expectancy of the population and changes in lifestyle. Studies on active principles of plant have motivated researchers to develop new antitumor agents that are specific and effective for treatment of neoplasms. Kaurane diterpenes are considered important compounds in the development of new and highly effective anticancer chemotherapeutic agents due to their cytotoxic properties in the induction of apoptosis. We evaluated the cytotoxic and apoptotic activity of the epimer of kaurenoic acid (EKA) isolated from the medicinal plant Croton antisyphiliticus (Euphorbiaceae) toward tumor cell lines HeLa and B-16 and normal fibroblasts 3T3. Based on analyses with the MTT test, EKA showed cytotoxic activity, with half maximal inhibitory concentration values of 59.41, 68.18 and 60.30 µg/mL for the B-16, HeLa and 3T3 cell lines, respectively. The assay for necrotic or apoptotic cells by differential staining showed induction of apoptosis in all three cell lines. We conclude that EKA is not selective between tumor and normal cell lines; the mechanism of action of EKA is induction of apoptosis, which is part of the innate mechanism of cell defense against neoplasia. PMID:23613246

Fernandes, V C; Pereira, S I V; Coppede, J; Martins, J S; Rizo, W F; Beleboni, R O; Marins, M; Pereira, P S; Pereira, A M S; Fachin, A L

2013-04-02

272

The epimer of kaurenoic acid from Croton antisyphiliticus is cytotoxic toward B-16 and HeLa tumor cells through apoptosis induction.  

UK PubMed Central (United Kingdom)

Cancer has become the leading cause of death in developing countries due to increased life expectancy of the population and changes in lifestyle. Studies on active principles of plant have motivated researchers to develop new antitumor agents that are specific and effective for treatment of neoplasms. Kaurane diterpenes are considered important compounds in the development of new and highly effective anticancer chemotherapeutic agents due to their cytotoxic properties in the induction of apoptosis. We evaluated the cytotoxic and apoptotic activity of the epimer of kaurenoic acid (EKA) isolated from the medicinal plant Croton antisyphiliticus (Euphorbiaceae) toward tumor cell lines HeLa and B-16 and normal fibroblasts 3T3. Based on analyses with the MTT test, EKA showed cytotoxic activity, with half maximal inhibitory concentration values of 59.41, 68.18 and 60.30 µg/mL for the B-16, HeLa and 3T3 cell lines, respectively. The assay for necrotic or apoptotic cells by differential staining showed induction of apoptosis in all three cell lines. We conclude that EKA is not selective between tumor and normal cell lines; the mechanism of action of EKA is induction of apoptosis, which is part of the innate mechanism of cell defense against neoplasia.

Fernandes VC; Pereira SI; Coppede J; Martins JS; Rizo WF; Beleboni RO; Marins M; Pereira PS; Pereira AM; Fachin AL

2013-01-01

273

Identification and characterization of a DNA primase activity present in herpes simplex virus type 1-infected HeLa cells  

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A novel DNA primase activity has been identified in HeLa cells infected with herpes simplex virus type 1 (HSV-1). Such an activity has not been detected in mock-infected cells. The primase activity coeluted with a portion of HSV-1 DNA polymerase from single-stranded DNA agarose columns loaded with high-salt extracts derived from infected cells. This DNA primase activity could be distinguished from host HeLa cell DNA primase by several criteria. First, the pH optimum of the HSV primase was relatively broad and peaked at 8.2 to 8.7 pH units. Second, freshly isolated HSV DNA primase was less salt sensitive than the HeLa primase. Third, antibodies raised against individual peptides of the calf thymus DNA polymerase:primase complex cross-reacted with the HeLa primase but did not react with the HSV DNA primase. Fourth, freshly prepared HSV DNA primase appeared to be associated with the HSV polymerase, but after storage at 4{degree}C for several weeks, the DNA primase separated from the viral DNA polymerase. This free DNA primase had an apparent molecular size of approximately 40 kilodaltons, whereas free HeLa DNA primase had an apparent molecular size of approximately 110 kilodaltons. On the basis of these data, the authors believe that the novel DNA primase activity in HSV-infected cells may be virus coded and that this enzyme represents a new and important function involved in the replication of HSV DNA.

Holmes, A.M.; Wietstock, S.M.; Ruyechan, W.T. (Uniformed Services Univ. of the Health Sciences, Bethesda, MD (USA))

1988-03-01

274

Identification and characterization of a DNA primase activity present in herpes simplex virus type 1-infected HeLa cells  

International Nuclear Information System (INIS)

[en] A novel DNA primase activity has been identified in HeLa cells infected with herpes simplex virus type 1 (HSV-1). Such an activity has not been detected in mock-infected cells. The primase activity coeluted with a portion of HSV-1 DNA polymerase from single-stranded DNA agarose columns loaded with high-salt extracts derived from infected cells. This DNA primase activity could be distinguished from host HeLa cell DNA primase by several criteria. First, the pH optimum of the HSV primase was relatively broad and peaked at 8.2 to 8.7 pH units. Second, freshly isolated HSV DNA primase was less salt sensitive than the HeLa primase. Third, antibodies raised against individual peptides of the calf thymus DNA polymerase:primase complex cross-reacted with the HeLa primase but did not react with the HSV DNA primase. Fourth, freshly prepared HSV DNA primase appeared to be associated with the HSV polymerase, but after storage at 4 degree C for several weeks, the DNA primase separated from the viral DNA polymerase. This free DNA primase had an apparent molecular size of approximately 40 kilodaltons, whereas free HeLa DNA primase had an apparent molecular size of approximately 110 kilodaltons. On the basis of these data, the authors believe that the novel DNA primase activity in HSV-infected cells may be virus coded and that this enzyme represents a new and important function involved in the replication of HSV DNA

1988-01-01

275

Chromatin assembly factor 1 (CAF-1) colocalizes with replication foci in HeLa cell nuclei.  

UK PubMed Central (United Kingdom)

In the S-phase of the eukaryotic cell cycle, newly replicated DNA is assembled into chromatin. I used indirect immunofluorescence microscopy to localize the sites of chromatin assembly in respect to DNA replication. Replication foci in the nuclei of permeabilized HeLa cells were labeled by incorporation of biotin-16-dUTP and detected by fluorescent streptavidin. Prelabeling of replication foci in vivo with bromodeoxyuridine showed that replication in permeabilized cells proceeds at preexisting replication forks. The localization of chromatin assembly factor 1 (CAF-1) was determined with subunit-specific monoclonal antibodies. CAF-1 is not detectable in mitotic cells and is detectable only at background levels in about 60% of all interphase nuclei. The other interphase nuclei show an intense punctate immunostaining of CAF-1. These sites of CAF-1 colocalize with replication foci during all stages of the S-phase. No other discrete sites of CAF-1 are observed. Human replication protein A (RPA) colocalizes with these replication/chromatin assembly sites. In addition, extra nuclear sites of RPA are observed that probably represent prereplication foci, poised for initiation of DNA replication.

Krude T

1995-10-01

276

Multiple origins of spontaneously arising micronuclei in HeLa cells: Direct evidence from long-term live cell imaging  

Energy Technology Data Exchange (ETDEWEB)

Although micronuclei (MNi) are extensively used to evaluate genotoxic effects and chromosome instability, the most basic issue regarding their origins has not been completely addressed due to limitations of traditional methods. Recently, long-term live cell imaging was developed to monitor the dynamics of single cell in a real-time and high-throughput manner. In the present study, this state-of-the-art technique was employed to examine spontaneous micronucleus (MN) formation in untreated HeLa cells. We demonstrate that spontaneous MNi are derived from incorrectly aligned chromosomes in metaphase (displaced chromosomes, DCs), lagging chromosomes (LCs) and broken chromosome bridges (CBs) in later mitotic stages, but not nuclear buds in S phase. However, most of bipolar mitoses with DCs (91.29%), LCs (73.11%) and broken CBs (88.93%) did not give rise to MNi. Our data also show directly, for the first time, that MNi could originate spontaneously from (1) MNi already presented in the mother cells; (2) nuclear fragments that appeared during mitosis with CB; and (3) chromosomes being extruded into a minicell which fused with one of the daughter cells later. Quantitatively, most of MNi originated from LCs (63.66%), DCs (10.97%) and broken CBs (9.25%). Taken together, these direct evidences show that there are multiple origins for spontaneously arising MNi in HeLa cells and each mechanism contributes to overall MN formation to different extents.

Rao Xiaotang; Zhang Yingyin; Yi Qiyi; Hou Heli; Xu Bo; Chu Liang; Huang Yun; Zhang Wenrui [Laboratory of Molecular and Cell Genetics, Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027 (China); Fenech, Michael [CSIRO Human Nutrition, PO Box 10041, Adelaide BC, Adelaide, SA 5000 (Australia); Shi Qinghua [Laboratory of Molecular and Cell Genetics, Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Hefei, Anhui 230027 (China)], E-mail: qshi@ustc.edu.cn

2008-11-10

277

Discrete foci containing RNAse A are found in nucleoli of HeLa cells after aging in culture  

Directory of Open Access Journals (Sweden)

Full Text Available We have studied by means of ultrastructural immunocytochemistry the localization of RNase A in nuclei of HeLa cells in control conditions and following cell ageing in culture. We have found that roundish, electron dense foci, which contain a significant amount of RNAse A, can be detected within nucleoli of aged cells. These bodies also contain RNA and lack ribosomal S3 proteins, and may represent either simple storage sites or areas where RNA degradation takes place.

M. Costanzo; B. Cisterna; O. O. Zharskaya; O. V. Zatsepina; M. Biggiogera

2011-01-01

278

Radiation-Induced DNA Labelling in G1 Phase in HeLa-S3 Cells  

International Nuclear Information System (INIS)

The primary aim of the experiments was to examine in HeLa-S3 tissue culture cells the effect of a single dose of X-irradiation on the rate of entry of cells into synthesizing DNA. To determine the rate at which cells enter DNA synthesis, different tracer techniques m a y be used. In this study, autoradiography with 3H-cytidine was chosen. 3H-cytidine enters a relatively large cellular precursor pool and can be utilized from there for DNA synthesis for at least one generation time. Using this technique, one observes autoradiographically the rise in the relative number of DNA-labelled cells as a function of the proliferative rate with time after flash-exposure of the cells to 3H-cytidine. Tracing the influx of DNA precursor from the cellular pool into DNA is physiological and eliminates the requirement of handling the cultures during experiment. Moreover, the effects on various phases of the cell cycle maybe analysed without having the cultures in a synchronous state of growth. After X-irradiation with 500 and 1000 R, an immediate mitotic delay was observed, which recovered between 16 and 24 hours after irradiation. Approximately 10% of the cell population was induced into synthesizing DNA. The effect occurred within 2 hours after irradiation. This increase of the labelling index was not due to changes in the autoradiographic efficiency, as was observed from measurements by interference microscopy, but to nuclear thickness and total nuclear mass. Analysis of the labelling index curves indicated that mainly cells of the latter part of the G1 phase were induced to synthesize DNA by irradiation. The rate of transition of G1 cells into S-phase following the initial effect was near normal and indistinguishable from the control values, at least for 8 hours after irradiation. (author)

1968-01-01

279

NMK-TD-100, a Novel Microtubule Modulating Agent, Blocks Mitosis and Induces Apoptosis in HeLa Cells by Binding to Tubulin  

Science.gov (United States)

Thiadiazoles are one of the most widely utilized agents in medicinal chemistry, having a wide range of pharmacologic activity. Microtubules (MTs) have always remained a sought-after target in rapidly proliferating cancer cells. We screened for the growth inhibitory effect of synthetic 5-(3-indolyl)-2-substituted-1,3,4-thiadiazoles on cancer cells and identified NMK-TD-100, as the most potent agent. Cell viability experiments using human cervical carcinoma cell line (HeLa cells) indicated that the IC50 value was 1.42±0.11 µM for NMK-TD-100 for 48 h treatment. In further study, we examined the mode of interaction of NMK-TD-100 with tubulin and unraveled the cellular mechanism responsible for its anti-tumor activity. NMK-TD-100 induced arrest in mitotic phase of cell cycle, caused decline in mitochondrial membrane potential and induced apoptosis in HeLa cells. Immunofluorescence studies using an anti-?-tubulin antibody showed a significant depolymerization of the interphase microtubule network and spindle microtubule in HeLa cells in a concentration-dependent manner. However, the cytotoxicity of NMK-TD-100 towards human peripheral blood mononuclear cells (PBMC) was lower compared to that in cancer cells. Polymerization of tissue purified tubulin into microtubules was inhibited by NMK-TD-100 with an IC50 value of 17.5±0.35 µM. The binding of NMK-TD-100 with tubulin was studied using NMK-TD-100 fluorescence enhancement and intrinsic tryptophan fluorescence of tubulin. The stoichiometry of NMK-TD-100 binding to tubulin is 1:1 (molar ratio) with a dissociation constant of ~1 µM. Fluorescence spectroscopic and molecular modeling data showed that NMK-TD-100 binds to tubulin at a site which is very near to the colchicine binding site. The binding of NMK-TD-100 to tubulin was estimated to be ~10 times faster than that of colchicine. The results indicated that NMK-TD-100 exerted anti-proliferative activity by disrupting microtubule functions through tubulin binding and provided insights into its potential of being a chemotherapeutic agent.

Bhattacharya, Surela; Kumar, N. Maruthi; Ganguli, Arnab; Tantak, Mukund P.; Kumar, Dalip; Chakrabarti, Gopal

2013-01-01

280

Cytochalasin B induces apoptosis through the mitochondrial apoptotic pathway in HeLa human cervical carcinoma cells.  

Science.gov (United States)

Cytochalasin B (CB) is a cell-permeable mycotoxin. It inhibits cytoplasmic division by blocking the formation of contractile microfilaments, it inhibits cell movement and induces nuclear extrusion. In the present study, we investigated the anticancer activity of CB in HeLa human cervical carcinoma cells. CB showed significant cytotoxicity, with an IC50 of 7.9 µM, in a WST-8 assay and significantly inhibited cell proliferation. Furthermore, results from Annexin V-FITC/propidium iodide double-staining indicated that CB induced early apoptosis of HeLa cells in a time-dependent manner. The cells exhibited apoptotic morphology, including cell shrinkage and nuclear condensation. CB induced cell cycle arrest at the S phase. We also observed inhibition of DNA replication in a [3H]-thymidine incorporation assay. Furthermore, CB induced a time-dependent increase in reactive oxygen species and a decrease in mitochondrial membrane potential. Western blot analysis showed an increase in levels of mitochondrial factors Bax and Bcl-2, which was followed by activation of caspase-9 and -3. These results suggested that CB induced apoptosis via a mitochondrial-dependent pathway in HeLa cells. PMID:23863920

Hwang, Jiyoung; Yi, Myeongjin; Zhang, Xin; Xu, Yi; Jung, Jee H; Kim, Dong-Kyoo

2013-07-17

 
 
 
 
281

Ultrastructural effects of two phthalocyanines in CHO-K1 and HeLa cells after laser irradiation  

Directory of Open Access Journals (Sweden)

Full Text Available The effects of Photodynamic Therapy using 2nd generation photosensitizers have been widely investigated aiming clinical application treatment of solid neoplasms. In this work, ultrastructure changes caused by the action of two 2nd generation photosensitizers and laser irradiation on CHO-K1 and HeLa (neoplastic) cells were analyzed by transmission electron microscopy. Aluminum phthalocyanine chloride, aluminum phthalocyanine tetrasulfonate chloride and radiation from a semiconductor laser at a fluency of 0.5 J/cm² (Power=26mW; l=670nm) were used. The results showed induction of apoptosis. Such alterations where observed in HeLa but not in CHO-K1 cells after Aluminum phthalocyanine tetrasulfonate chloride (AlPcS4) photodynamic treatment. The Aluminum phthalocyanine chloride (AlPc) photodynamic treatment induced necrosis on the neoplastic cell line, and cytoplasm and nuclear alterations on the normal cell line.

Marcelo de CastroPazos; Cristina Pacheco-Soares; Newton Soares da Silva; Renato Augusto DaMatta; Marcos Tadeu T. Pacheco

2003-01-01

282

Ultrastructural effects of two phthalocyanines in CHO-K1 and HeLa cells after laser irradiation  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english The effects of Photodynamic Therapy using 2nd generation photosensitizers have been widely investigated aiming clinical application treatment of solid neoplasms. In this work, ultrastructure changes caused by the action of two 2nd generation photosensitizers and laser irradiation on CHO-K1 and HeLa (neoplastic) cells were analyzed by transmission electron microscopy. Aluminum phthalocyanine chloride, aluminum phthalocyanine tetrasulfonate chloride and radiation from a sem (more) iconductor laser at a fluency of 0.5 J/cm² (Power=26mW; l=670nm) were used. The results showed induction of apoptosis. Such alterations where observed in HeLa but not in CHO-K1 cells after Aluminum phthalocyanine tetrasulfonate chloride (AlPcS4) photodynamic treatment. The Aluminum phthalocyanine chloride (AlPc) photodynamic treatment induced necrosis on the neoplastic cell line, and cytoplasm and nuclear alterations on the normal cell line.

de CastroPazos, Marcelo; Pacheco-Soares, Cristina; Soares da Silva, Newton; DaMatta, Renato Augusto; Pacheco, Marcos Tadeu T.

2003-12-01

283

Ruthenium (II) polypyridyl complexes stabilize the bcl-2 promoter quadruplex and induce apoptosis of Hela tumor cells.  

Science.gov (United States)

In the present study, the interaction between GC-rich sequence of bcl-2 gene P1 promoter (Pu39) and two ruthenium (II) polypyridyl complexes, [Ru(bpy)?(tip)]²? (1) and [Ru(phen)?(tip)]²? (2), was investigated by UV-Visible, fluorescence spectroscopy, circular dichroism, fluorescence resonance energy transfer melting assay and polymerase chain reaction stop assay. Those experimental results indicated that the two complexes can effectively stabilize the G-quadruplex of Pu39. It was found that the complex 2 exhibited greater cytotoxic activity than 1 against human Hela cells and can enter into Hela cells in a short period of time to effectively induce apoptosis of cells. Further experiments found that complexes 1 and 2 had as potent inhibitory effects on ECV-304 cell migration as suramin. Those noteworthy results provide new insights into the development of anticancer agents for targeting G-quadruplex DNA. PMID:23543383

Wang, Chuan; Yu, Qianqian; Yang, Licong; Liu, Yanyu; Sun, Dongdong; Huang, Yongchao; Zhou, Yanhui; Zhang, Qianling; Liu, Jie

2013-03-30

284

Ruthenium (II) polypyridyl complexes stabilize the bcl-2 promoter quadruplex and induce apoptosis of Hela tumor cells.  

UK PubMed Central (United Kingdom)

In the present study, the interaction between GC-rich sequence of bcl-2 gene P1 promoter (Pu39) and two ruthenium (II) polypyridyl complexes, [Ru(bpy)?(tip)]²? (1) and [Ru(phen)?(tip)]²? (2), was investigated by UV-Visible, fluorescence spectroscopy, circular dichroism, fluorescence resonance energy transfer melting assay and polymerase chain reaction stop assay. Those experimental results indicated that the two complexes can effectively stabilize the G-quadruplex of Pu39. It was found that the complex 2 exhibited greater cytotoxic activity than 1 against human Hela cells and can enter into Hela cells in a short period of time to effectively induce apoptosis of cells. Further experiments found that complexes 1 and 2 had as potent inhibitory effects on ECV-304 cell migration as suramin. Those noteworthy results provide new insights into the development of anticancer agents for targeting G-quadruplex DNA.

Wang C; Yu Q; Yang L; Liu Y; Sun D; Huang Y; Zhou Y; Zhang Q; Liu J

2013-06-01

285

Cell proliferation and viability of HELA and HEP-2P neoplastic cultures in vitro treated with new polyphenolic or polysaccharidic biopreparations of vegetable and fungal origin  

Directory of Open Access Journals (Sweden)

Full Text Available The in vitro action of some new autochthonous polysaccharidic and polyphenolic biopreparations, specifically extracted from fungal and vegetable sources, upon the proliferation and viability of the HeLa and Hep-2p cancerous cells was investigated. The significant perturbation of the mitosis process of the tumoral cells, as well as the profound diminution of the cell viability of the neoplastic cell cultures argue the behaviour of some polysaccharidic and polyphenolic extracts as in vitro active cytostatic and cytotoxic agents. This primary characterization of some natural fungal or vegetable extracts as cytostatic and/or cytotoxic agents offers the informational background for the introduction of those biopreparations in the in vivo antitumoral screening program on different experimental tumoral systems.

Pincu Rotinberg; Cosmin Mihai; Daniela Gherghel; Elena Truta; Gabriela Capraru; Ruxandra Cretu; Ion Neacsu; Hellen Rotinberg

2008-01-01

286

Synthesis of polyfunctionalized piperidone oxime ethers and their cytotoxicity on HeLa cells.  

Science.gov (United States)

A series of twenty 2,6-diarylpiperidin-4-one O-methyloximes were synthesized with fluoro/chloro/bromo/methyl/methoxy/ethoxy/isopropyl substituents on various positions of the phenyl at C-2 and C-6 in association with/without methyl substituent on the secondary amino group and methyl/ethyl/isopropyl substituents on the active methylene centers. Regardless of their substitution all compounds predominantly exist in the chair conformation except 3m, which adopts a twist-boat conformation. All the synthesized compounds were evaluated for their in vitro antiproliferative activity against human cervical carcinoma (HeLa) cell line. The cytotoxicity of the test compounds was determined by measuring the number of live cells after 24 h of treatment by MTT assay method. This preliminary SAR suggests some lead molecules 3c-f, 3j-k, 4d-g, and 4i with a scope of further structural optimization of the piperidone pharmacophore toward the development of anticancer drug synthesis. PMID:21983445

Parthiban, Paramasivam; Pallela, Ramjee; Kim, Se-Kwon; Park, Dong Ho; Jeong, Yeon Tae

2011-09-21

287

Synthesis of polyfunctionalized piperidone oxime ethers and their cytotoxicity on HeLa cells.  

UK PubMed Central (United Kingdom)

A series of twenty 2,6-diarylpiperidin-4-one O-methyloximes were synthesized with fluoro/chloro/bromo/methyl/methoxy/ethoxy/isopropyl substituents on various positions of the phenyl at C-2 and C-6 in association with/without methyl substituent on the secondary amino group and methyl/ethyl/isopropyl substituents on the active methylene centers. Regardless of their substitution all compounds predominantly exist in the chair conformation except 3m, which adopts a twist-boat conformation. All the synthesized compounds were evaluated for their in vitro antiproliferative activity against human cervical carcinoma (HeLa) cell line. The cytotoxicity of the test compounds was determined by measuring the number of live cells after 24 h of treatment by MTT assay method. This preliminary SAR suggests some lead molecules 3c-f, 3j-k, 4d-g, and 4i with a scope of further structural optimization of the piperidone pharmacophore toward the development of anticancer drug synthesis.

Parthiban P; Pallela R; Kim SK; Park DH; Jeong YT

2011-11-01

288

Effects of a tumor promoter on phospholipid metabolism in HeLa cells  

Energy Technology Data Exchange (ETDEWEB)

The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) caused a marked stimulation of inorganic (/sup 32/P)orthophosphate incorporation into HeLa-cell phosphatidylcholine (PC), phosphatidylethanolamine (PE), and lysophosphatidylethanolamine. The increased incorporation of inorganic (/sup 32/P)orthophosphate into PE and lysophosphatidylethanolamine in the presence of TPA was not associated with an increase in PE synthesis as detected by the incorporation of (/sup 3/H)serine or (/sup 3/H)ethanolamine. The PC-specific exchange protein from beef liver was used to insert PC labeled with (/sup 3/H)choline, inorganic (/sup 32/P)orthophosphate, or (/sup 14/C)arachidonic acid plus (/sup 3/H)palmitic acid into the outer monolayer of intact HeLa cell membranes. Radioactivity from the latter two compounds was rapidly incorporated into PE and lysophosphatidylethanolamine; the incorporation was stimulated by TPA. It was concluded that TPA stimulated the formation of PE by base exchange between ethanolamine and PC.

Guy, G.R.; Murray, A.W.

1983-11-01

289

Chlamydia trachomatis serovar L2 induces protein tyrosine phosphorylation during uptake by HeLa cells.  

DEFF Research Database (Denmark)

Birkelund S , Johnsen H , Christiansen G . Institute of Medical Microbiology, University of Aarhus, Denmark. Chlamydia trachomatis L2 is an obligate intracellular microorganism with a unique biphasic life cycle. The extracellular form, the elementary body (EB), is infectious but metabolically inactive. Attachment of EBs to host cells is medicated by a heparan sulfate-like glycosaminoglycan. Following attachment, the EB is internalized within a membrane-bound vesicle, and during the first 8 h of infection the vesicles are transported to a perinuclear location where they aggregate and fuse. By use of a monoclonal antibody against phosphotyrosine, we showed that three classes of proteins are tyrosine phosphorylated: a triple band of 68, 66, and 64 kDa, a 97-kDa band, and a 140-kDa band. The phosphorylation could be detected by immunoblotting from 15 min after infection of HeLa cells. We followed the movement of the EBs and the tyrosine phosphorylation of proteins by double-labelling immunofluorescence microscopy with the same monoclonal anti-phosphotyrosine antibody and a polyclonal antibody against the C. trachomatis L2 outer membrane complex. During the first 8 h of infection, the phosphorylation colocalized with EBs. Sixteen hours after infection, EBs have reorganized to the replicating reticulate bodies, forming an inclusion. At this time, phosphorylation was seen as dotted spots in the periphery of the inclusion.

Birkelund, Svend; Johnsen, H

1994-01-01

290

Enhanced reactivation of UV-irradiated adenovirus 2 in HeLa cells treated with non-mutagenic chemical agents  

International Nuclear Information System (INIS)

Treatment of HeLa cells with ethanol and sodium arsenite, compounds which are known to elicit the heat-shock response, before infection with UV-irradiated adenovirus 2 has been found to result in the enhanced reactivation of the damaged virus in a manner similar to that obtained by pre-irradiation or heating of the cells. Enhanced reactivation may be the result of the inhibition of DNA synthesis caused by these agents since hydroxyurea also produced a significant enhancement. (orig.).

1985-01-01

291

Antioxidant Potential of Ulva rigida Extracts: Protection of HeLa Cells Against H2O2 Cytotoxicity.  

UK PubMed Central (United Kingdom)

The rising demand for natural antioxidants instead of synthetic materials, especially in biomedical applications, has led to increased interest in the search for bioactive compounds with potent antioxidant activity. In the present study, we tested the antioxidant effect of both a crude extract and an ethanol precipitate of Ulva rigida in HeLa cells exposed to hydrogen peroxide (H2O2). HeLa cells treated with H2O2 (1 mmol l(-1) for 3 h) exhibited significant damage to their morphology, a significant decrease in cell survival, and a remarkable leakage of lactate dehydrogenase (LDH). However, the co-exposure of cells to H2O2 and the crude extract or ethanol precipitate of U. rigida induced fewer morphological cytotoxic effects, a significant increase in cell viability, and a significant decrease in LDH release. Biochemical studies have demonstrated that U. rigida extracts have a strong radicalscavenging activity and contain protein, sugar, and phenolic content. The overall results suggest that U. rigida extracts protect HeLa cells from death induced by oxidative stress, and it is likely that these effects are related to the phenolic, protein, and polysaccharide compounds contained in this alga. Hence, U. rigida can be used to treat diseases ascribed to oxidative disorders.

Mezghani S; Bourguiba I; Hfaiedh I; Amri M

2013-09-01

292

Radiosensitizing effect of gold nanoparticles in carbon ion irradiation of human cervical cancer cells  

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Noble metal nanoparticles have received considerable attention in biotechnology for their role in bio sensing due to surface plasmon resonance, medical diagnostics due to better imaging contrast and therapy. The radiosensitization effect of gold nanoparticles (AuNP) has been gaining popularity in radiation therapy of cancer cells. The better depth dose profile of energetic ion beam proves its superiority over gamma radiation for fighting against cancer. In the present work, the glucose capped gold nanoparticles (Glu-AuNP) were synthesised and internalized in the HeLa cells. Transmission electron microscopic analysis of ultrathin sections of Glu-AuNP treated HeLa cells confirmed the internalization of Glu-AuNPs. Control HeLa cells and Glu-AuNp treated HeLa cells were irradiated at different doses of 62 MeV 12C ion beam (LET - 290keV/{mu}m) at BIO beam line of using 15UD Pelletron accelerator at Inter University Accelerator Centre, New Delhi, India. The survival fraction was assessed by colony forming assay which revealed that the dose of carbon ion for 90% cell killing in Glu-AuNP treated HeLa cells and control HeLa cells are 2.3 and 3.2 Gy respectively. This observation shows {approx} 28% reduction of {sup 12}C{sup 6+} ion dose for Glu-AuNP treated HeLa cells as compared to control HeLa cells.

Kaur, Harminder; Avasthi, D. K.; Pujari, Geetanjali; Sarma, Asitikantha [Inter University Accelerator Centre, Aruna Asaf Ali Marg, Post box-10502, New Delhi-110067 (India)

2013-07-18

293

HeLa cell tumor response to 60Co, Cs-137, Cf-252 radiations and cisplatin chemotherapy in nude mice  

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HeLa cells were implanted into athymic nude mice from tissue culture and solid tumors established (HeLa cell tumor or HCT). Large cell numbers of 1 X 10/sup 7/ were required to obtain consistent and progressive growth, and tumor growth followed a Gompertzian mode. Irradiation studies were carried out using acute Cobalt-60 (60Co), low-dose-rate (LDR) Cs-137 and LDR Cf-252. Cf-252, a neutron-emitting radioisotope, produced an immediate tumor shrinkage and regression response after a dose of 279 cGy. Acute 60Co or LDR Cs-137 irradiation with 1000 cGy had little effect on the HCT. After a dose of 2000 cGy of 60Co radiation tumor shrinkage followed a latent period of approximately 5 days. Cisplatin had no effect on the HCT in nude mice in stationary or late exponential growth.

Maruyama, Y.; Feola, J.M.; Beach, J.L.

1984-07-15

294

Effect of 12-O-tetradecanoylphorbol-13-acetate (TPA) on phosphatidylethanolamine metabolism in HeLa cells  

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The potent tumor promoter, TPA, exerts its earliest effects at the plasma membrane. Recent findings have shown that TPA stimulates a phospholipase C-mediated turnover of phosphatidyl-choline in several different cell types. The present study was undertaken to investigate whether TPA elicits a similar effect on the phosphatidylethanolamine (PE) pool of HeLa cells. Three different series of experiments were performed. First, in HeLa cells pulse-labeled with (/sup 3/H)ethanolamine, TPA stimulated a 5-fold release of aqueous radiolabeled products into the extra-cellular medium after a 1-hour incubation. Second, when (/sup 3/H)ethanolamine and TPA were added simultaneously to the cells, TPA stimulated a 2-fold incorporation of radiolabel into the cellular PE pool. In both the release and incorporation of (/sup 3/H)ethanolamine, TPA had no significant effect on PE mass. Finally, when HeLa cells were incubated with exogenous 1-radyl-2-acyl-sn-glycero-3-phospho-(/sup 3/H)ethanolamine, TPA stimulated the formation of an aqueous radiolabeled product in the medium, which was identified as phosphoethanolamine. These results provide evidence that TPA stimulates a phospholipase C-mediated turnover of PE.

Mueller, H.W.; Vance, D.E.

1986-05-01

295

Multiple signal transduction pathways in okadaic acid induced apoptosis in HeLa cells  

International Nuclear Information System (INIS)

[en] Okadaic acid (OA) is the major component of diarrhetic shell fish poisoning toxins and a potent inhibitor of protein phosphatase 1 and 2A. We investigated the signal transduction pathways involved in OA induced cell death in HeLa cells. OA induced cytotoxicity and apoptosis at IC50 of 100 nM. OA treatment resulted in time dependent increase in reactive oxygen species and depleted intracellular glutathione levels. Loss of mitochondrial membrane permeability led to translocation of bax, cytochrome-c and AIF from mitochondria to cytosol. The cells under fluorescence microscope showed typical apoptotic morphology with condensed chromatin, and nuclear fragmentation. We investigated the mitochondrial-mediated caspase cascade. The time dependent activation and cleavage of of bax, caspases-8, 10, 9, 3 and 7 was observed in Western blot analysis. In addition to caspase-dependent pathway AIF mediated caspase-independent pathway was involved in OA mediated cell death. OA also caused time dependent inhibition of protein phosphatase 2A activity and phosphorylation of p38 and p42/44 MAP kinases. Inhibitor studies with Ac-DEVO-CHO and Z-VAD-FMK could not prevent the phosphorylation of p38 and p42/44 MAP kinases. Our experiments with caspase inhibitors Ac-DEVD-CHO, Z-IETD-FMK and Z-VAD-FMK inhibited capsase-3, 8 cleavages but did not prevent OA-induced apoptosis and DNA fragmentation. Similarly, pretreatment with cyclosporin-A and N-acetylcysteine could not prevent the DNA fragmentation. In summary, the results of our study show that OA induces multiple signal transduction pathways acting either independently or simultaneously leading to apoptosis

2009-02-04

296

Fractionation of HeLa cell nuclear extracts reveals minor small nuclear ribonucleoprotein particles  

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Upon chromatographic fractionation of HeLa cell nuclear extracts, small RNAs of 145 and 66/65 nucleotides, respectively, were detected that are distinct from the abundant small RNAs present in the extract. These RNAs are precipitated by antibodies directed against the trimethylguanosine cap structure, characteristic for small nuclear RNAs (snRNAs) of the U type. The RNAs of 145 and 66/65 nucleotides appear to be associated with at least one of the proteins common to the major small nuclear ribonucleoprotein particles U1 to U6, since they are specifically bound by anti-Sm antibodies. These criteria characterize the RNAs that are 145 and 66/65 nucleotides in length as U-type snRNAs. Upon gel filtration, the RNAs are found within particles of molecular weights approx. = 150,000 and 115,000 respectively. The RNA of 145 nucleotides represents a different minor snRNA, designated U11, whereas the RNA of 66/65 nucleotides may correspond to either mammalian U7 or U10 RNA.

Kroemer, A.

1987-12-01

297

Fractionation of HeLa cell nuclear extracts reveals minor small nuclear ribonucleoprotein particles  

International Nuclear Information System (INIS)

Upon chromatographic fractionation of HeLa cell nuclear extracts, small RNAs of 145 and 66/65 nucleotides, respectively, were detected that are distinct from the abundant small RNAs present in the extract. These RNAs are precipitated by antibodies directed against the trimethylguanosine cap structure, characteristic for small nuclear RNAs (snRNAs) of the U type. The RNAs of 145 and 66/65 nucleotides appear to be associated with at least one of the proteins common to the major small nuclear ribonucleoprotein particles U1 to U6, since they are specifically bound by anti-Sm antibodies. These criteria characterize the RNAs that are 145 and 66/65 nucleotides in length as U-type snRNAs. Upon gel filtration, the RNAs are found within particles of molecular weights ? 150,000 and 115,000 respectively. The RNA of 145 nucleotides represents a different minor snRNA, designated U11, whereas the RNA of 66/65 nucleotides may correspond to either mammalian U7 or U10 RNA.

1987-01-01

298

Bisquinolinium compounds induce quadruplex-specific transcriptome changes in HeLa S3 cell lines  

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Full Text Available Abstract Background Guanosine rich sequences capable of forming G-quadruplex (G4) motifs are enriched near the gene transcription start site (TSS) in the human genome. When probed at the single gene level, G-quadruplex motifs residing in promoter regions show substantial effects on gene transcription. Moreover, further changes in transcription levels are noticed when G4-motifs are targeted with G-quadruplex-specific small molecules. Results Global studies concerning general changes of the transcriptome via targeting promoter-based G-quadruplex motifs are very limited and have so far only been carried out with compounds displaying weak selectivity for quadruplex sequences. Here we utilize two G-quadruplex-specific bisquinolinium derivatives PhenDC3 and 360A and investigate their effects on the expression of the HeLa S3 transcriptome. Our results show wide-spread changes in the transcriptome with specificity for genes with G-quadruplex motifs near their transcription start sites (TSS). Using real-time PCR we further confirmed the specificity of PhenDC3 and 360A as potent molecules to target G-quadruplex-regulated genes. Conclusions Specific effects on quadruplex-containing genes have been observed utilizing whole-transcriptome analysis upon treatment of cultured cells with quadruplex-selective bisquinolinium compounds.

Halder Rashi; Riou Jean-Francois; Teulade-Fichou Marie-Paule; Frickey Tancred; Hartig Jörg S

2012-01-01

299

MDM2 promotes the proteasomal degradation of p73 through the interaction with Itch in HeLa cells.  

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It has been shown that MDM2 inhibits the transcriptional and pro-apoptotic activities of p73 but does not promote its proteasomal degradation. In this study, we found that MDM2 indirectly induces the degradation of p73 through the interaction with Itch in HeLa cells. During adriamycin (ADR)-mediated apoptosis, p53 and p73 were induced to stabilize in association with a significant reduction of MDM2 and Itch, suggesting that, in addition to Itch, MDM2 could also be involved in the stability control of p73. As expected, forced expression of MDM2 resulted in a remarkable reduction of p73. MDM2-mediated degradation of p73 was inhibited by MG-132. Intriguingly, siRNA-mediated knockdown of Itch significantly attenuated the negative effect of MDM2 on p73. Additionally, MDM2 bound to Itch in HeLa cells but not in H1299 cells. Collectively, our present findings suggest that MDM2 promotes Itch-mediated degradation of p73 through the interaction with Itch in HeLa cells. PMID:21093410

Kubo, Natsumi; Okoshi, Rintaro; Nakashima, Kumiko; Shimozato, Osamu; Nakagawara, Akira; Ozaki, Toshinori

2010-11-17

300

18 S ribosomal RNA is degraded during ribosome maturation in irradiated HeLa cells  

International Nuclear Information System (INIS)

[en] The effects of ionizing radiation (137Cs) on processing of ribosomal RNA (rRNA) were studied by pulse-labeling HeLa S3 cells with [3H]uridine immediately prior to irradiation. The 45 S rRNA precursor, and its two major daughter species, 28 and 18 S rRNA, were separated by gel electrophoresis and the extent of radiolabel incorporation into each was determined at various times after irradiation. This approach permitted kinetic analysis of processing of the 45 S rRNA which had been predominantly synthesized (radiolabeled) prior to irradiation. Since they both derive from the same 45 S pre-rRNA transcript, 28 and 18 S rRNA are produced with a stoichiometry of 1:1, as observed in control cells in the present studies. However, within 1 h following 10 Gy an altered stoichiometry of 28 S:18 S rRNA was apparent, reaching 1.6:1 by 5-7 h following irradiation. This alteration was also observed following the higher dose of 20 Gy, but not following exposures of 5 Gy or less. The 18 S portion of the 45 S pre-rRNA is transcribed prior to the 28 S portion. Consequently, an increase in the 28 S/18 S ratio can only be due to degradation of the 18 S species during or after processing. This alteration may represent a response to radiation-induced growth arrest, by reducing the number of newly synthesized ribosomes that would otherwise be required for cell propagation

1989-01-01

 
 
 
 
301

ERK-1 MAP kinase prevents TNF-induced apoptosis through bad phosphorylation and inhibition of Bax translocation in HeLa Cells.  

UK PubMed Central (United Kingdom)

Extracellular signal-regulated kinase (ERK) 1/2 signaling is involved in tumor cell survival through the regulation of Bcl-2 family members. To explore this further and to demonstrate the central role of the mitochondria in the ERK1/2 pathway we used the HeLa cellular model where apoptosis was induced by tumor necrosis factor (TNF) and cycloheximide (CHX). We show that HeLa cells overexpressing ERK-1 displayed resistance to TNF and CHX. HeLa cells overexpressing a kinase-deficient form of ERK-1 (K71R) were more sensitive to TNF and CHX. In the ERK-1 cells, Bad was phosphorylated during TNF + CHX treatment. In the HeLa wt cells and in the K71R clones TNF and CHX decreased Bad phosphorylation. ERK-1 cells treated with TNF and CHX did not release cytochrome c from the mitochondria. By contrast, HeLa wt and K71R clones released cytochrome c. Bax did not translocate to the mitochondria in ERK-1 cells treated with TNF + CHX. Conversely, HeLa wt and K71R clones accumulated Bax in the mitochondria. In the HeLa wt cells and in both ERK-1 transfectants Bid was cleaved and accumulated in the mitochondria. The caspase-8 inhibitor IETD-FMK and the mitochondrial membrane permeabilization inhibitor bongkrekic acid (BK), partially prevented cell death by TNF + CHX. Anisomycin, a c-Jun N-terminal kinases activator, increased TNF-killing. The ERK-1 cells were resistant to TNF and anisomycin, whereas K71R clones resulted more sensitive. Our study demonstrates that in HeLa cells the ERK-1 kinase prevents TNF + CHX apoptosis by regulating the intrinsic mitochondrial pathway through different mechanisms. Inhibition of the intrinsic pathway is sufficient to almost completely prevent cell death.

Pucci B; Indelicato M; Paradisi V; Reali V; Pellegrini L; Aventaggiato M; Karpinich NO; Fini M; Russo MA; Farber JL; Tafani M

2009-12-01

302

ERK-1 MAP kinase prevents TNF-induced apoptosis through bad phosphorylation and inhibition of Bax translocation in HeLa Cells.  

Science.gov (United States)

Extracellular signal-regulated kinase (ERK) 1/2 signaling is involved in tumor cell survival through the regulation of Bcl-2 family members. To explore this further and to demonstrate the central role of the mitochondria in the ERK1/2 pathway we used the HeLa cellular model where apoptosis was induced by tumor necrosis factor (TNF) and cycloheximide (CHX). We show that HeLa cells overexpressing ERK-1 displayed resistance to TNF and CHX. HeLa cells overexpressing a kinase-deficient form of ERK-1 (K71R) were more sensitive to TNF and CHX. In the ERK-1 cells, Bad was phosphorylated during TNF + CHX treatment. In the HeLa wt cells and in the K71R clones TNF and CHX decreased Bad phosphorylation. ERK-1 cells treated with TNF and CHX did not release cytochrome c from the mitochondria. By contrast, HeLa wt and K71R clones released cytochrome c. Bax did not translocate to the mitochondria in ERK-1 cells treated with TNF + CHX. Conversely, HeLa wt and K71R clones accumulated Bax in the mitochondria. In the HeLa wt cells and in both ERK-1 transfectants Bid was cleaved and accumulated in the mitochondria. The caspase-8 inhibitor IETD-FMK and the mitochondrial membrane permeabilization inhibitor bongkrekic acid (BK), partially prevented cell death by TNF + CHX. Anisomycin, a c-Jun N-terminal kinases activator, increased TNF-killing. The ERK-1 cells were resistant to TNF and anisomycin, whereas K71R clones resulted more sensitive. Our study demonstrates that in HeLa cells the ERK-1 kinase prevents TNF + CHX apoptosis by regulating the intrinsic mitochondrial pathway through different mechanisms. Inhibition of the intrinsic pathway is sufficient to almost completely prevent cell death. PMID:19777442

Pucci, Bruna; Indelicato, Manuela; Paradisi, Valentina; Reali, Valentina; Pellegrini, Laura; Aventaggiato, Michele; Karpinich, Natalie O; Fini, Massimo; Russo, Matteo A; Farber, John L; Tafani, Marco

2009-12-01

303

Comparative experimental studies into radioimmunoscintigraphy using radioactive antibodies in animals with HeLa cell carcinomas and Yoshida sarcomas  

International Nuclear Information System (INIS)

TPA-positive and TPA-negative tumour-bearing animal systems (HeLa cell carcinomas in RNU rats and Yoshida sarcomas in Wistar rats) were examined to show that the method of scanning can well be used to visualise tumour tissue. In this connection, further attempts were made to shed light on the specifity of immunoscintigraphy in the search for tumour tissue. 125-Iodine-anti-TPA was found to be a specific carcinoma-seeking substance. The amount of antibodies accumulating in the tumour was multiplied by previous intravenous treatment of test animals with unspecific immunoglobulin. In control studies using 125-iodine-immunoglobulin the site of the carcinomatous tissue could not be determined with sufficient diagnostic accuracy. It was found that the discriminating power of radioimmunoscintigraphy using 125-iodine-anti-TPA is quite unrelated to an increased circulation in the proliferating carcinomatous tissue. For the detection of TPA in HeLa cell carcinomas anti-TPA PAP stains were prepared. Radionuclide studies using 125-iodine-anti-TPA were also useful in the visualisation of the Yoshida sarcoma, even though this scores negative on TPA. Here, the amounts of radioactivity accumulating in the tumour were smaller than with the HeLa cell carcinoma. Moreover, peak levels were measured after no less than one day, as compared to the five days required for HeLa cell tumours to reach maximum levels. This finding would appear to provide presumptive evidence that there are other, unspecific mechanisms of tumour selectivity. (orig/MG)

1988-01-01

304

Application of a FRET probe for Caspase-3 activation in living HeLa cells by sequentially treated cisplatin and TRAIL  

Science.gov (United States)

Caspase-3 is a kind of cysteine proteases that plays an important role in cell apoptosis. We have constructed a FRET (fluorescence resonance energy transfer) probe fused with ECFP (enhanced cyan fluorescence protein) and DsRed (Discosoma red fluorescent protein) with a linker containing a caspase-3 cleavage sequence (CCS, DEVD).It could be observed much change in fluorescence emission ratio when the probe was cleaved by caspase-3. Therefore, application of this probe we can real-time detected the activation of caspase-3. It was already confirmed that caspase-3 was activated in HeLa cells treated by cisplatin or TRAIL (Tumor necrosis factor (TNF)-related apoptosis-inducing ligand). In the present study, we detected the activation of caspase-3 during cisplatin or TRAIL induced apoptosis in living HeLa cells, and also observed the activation of caspase-3 caused by both cisplatin and TRAIL combined treatment. Our results demonstrated a synergistic effect between cisplatin and TRAIL. Cisplatin is one of the most broadly used drugs in the Clinical applications of cancer chemotherapy, and TRAIL, which belongs to the TNF family proteins, can selectively induce apoptosis in many transformed cells but not in normal cells. Therefore, TRAIL is a very valuably prospective utility as its potential tumor-specific cancer therapeutic. Most of anticancer drugs can induce apoptosis which mediated by the activation of caspase pathway. We can select the best synergistic effect group by our FRET probe. This finding would be useful in the design of treatment modalities for patients.

Lin, Juqiang; Zhang, Zhihong; Yi, Qiushi; Zeng, Shaoqun; Luo, Qingming

2006-03-01

305

RGDC functionalized titanium dioxide nanoparticles induce less damage to plasmid DNA but higher cytotoxicity to HeLa cells.  

Science.gov (United States)

In this paper, nano-TiO(2) was functionalized by different methods, and its genotoxicity and cytotoxicity were studied in detail. The genotoxicity of nano-TiO(2) was evaluated by observing its interactions with pUC19 plasmid DNA at a single molecule level using atomic force microscopy. The results show that with the assistance of UVA radiation, RGDC functionalized nano-TiO(2) induced less damage to plasmid DNA than unmodified ones. The HeLa cell-specific PDT effect was investigated by cytotoxicity assay correspondingly. RGDC-functionalized nano-TiO(2) shows the highest killing effect to HeLa cells with the assistance of UVA radiation. The reasons that cause the contradiction between genotoxicity and cytotoxicity were analyzed, and the molecular mechanisms of the PDT effects were discussed. The results show that the genotoxicity of nano-TiO(2) to plasmid DNA and its cytotoxicity to HeLa cells are related but also different. The RGDC functionalization is an effective method to increase the cytotoxicity of nano-TiO(2). PMID:23210915

Yin, Yuan; Zhu, Wei-Wei; Guo, Li-Ping; Yang, Ran; Li, Xin-Song; Jiang, Yong

2012-12-20

306

RGDC functionalized titanium dioxide nanoparticles induce less damage to plasmid DNA but higher cytotoxicity to HeLa cells.  

UK PubMed Central (United Kingdom)

In this paper, nano-TiO(2) was functionalized by different methods, and its genotoxicity and cytotoxicity were studied in detail. The genotoxicity of nano-TiO(2) was evaluated by observing its interactions with pUC19 plasmid DNA at a single molecule level using atomic force microscopy. The results show that with the assistance of UVA radiation, RGDC functionalized nano-TiO(2) induced less damage to plasmid DNA than unmodified ones. The HeLa cell-specific PDT effect was investigated by cytotoxicity assay correspondingly. RGDC-functionalized nano-TiO(2) shows the highest killing effect to HeLa cells with the assistance of UVA radiation. The reasons that cause the contradiction between genotoxicity and cytotoxicity were analyzed, and the molecular mechanisms of the PDT effects were discussed. The results show that the genotoxicity of nano-TiO(2) to plasmid DNA and its cytotoxicity to HeLa cells are related but also different. The RGDC functionalization is an effective method to increase the cytotoxicity of nano-TiO(2).

Yin Y; Zhu WW; Guo LP; Yang R; Li XS; Jiang Y

2013-01-01

307

Antibodies against recombinant catalytic domain of lethal toxin of Clostridium sordellii neutralize lethal toxin toxicity in HeLa cells.  

UK PubMed Central (United Kingdom)

Lethal toxin of Clostridium sordellii (MLD 150 ng/kg) is one of the most potent Clostridial toxins and is responsible for most of the diseases including sudden death syndrome in cattle, sheep and toxic shock syndrome, necrotizing faciitis, neonatal omphalitis and gangrene in humans. Lethal toxin (TcsL) is a single chain protein of about 270 kDa. In the present study, 1.6 kb DNA fragment encoding for the catalytic domain of TcsL was PCR amplified, cloned in pQE30 UA vector and expressed in E. coli SG 13009. The expression of recombinant lethal toxin protein (rTcsL) was optimized and it was purified under native conditions using a single step Ni-NTA affinity chromatography. The purified recombinant protein was used for the production of polyclonal antibodies in mice and rabbit. The raised antibodies reacted specifically with the purified rTcsL and intact native lethal toxin on Western blot. The biological activity of the recombinant protein was tested in HeLa cells where it showed the cytotoxicity. Further, the polyclonal antibodies were used for in-vitro neutralization of purified rTcsL, acid precipitated C. sordellii and C. difficile native toxins in HeLa cells. Mice and rabbit anti-rTcsL sera effectively neutralized the cytotoxicity of rTcsL and C. sordellii native toxin but it did not neutralize the cytotoxicity of C. difficile toxin in HeLa cells.

Arya P; Ponmariappan S; Singh L; Prasad GB

2013-02-01

308

Cytotoxicity studies of microwave assisted natural product extracts in HeLa and MCF-7 cell lines  

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Full Text Available Microwave assisted extraction of leaves and stems of seven tropical plants viz. Ficus hispida, Andrographis paniculata, Alistonia scholaris, Aporusa lindlyeyana, Melastoma malabathicum, Adenon indicum and Alternanthera dentata was carried out. Optimum extraction conditions were 600 W microwave power, 6 min extraction time (spread over two extraction cycles of 3 min each) and 10 min pre-leaching time using powdered stem and leaves samples. The two solvents employed were 90% ethanol and hexane with 10:1 (mL/g) as the solvent to material loading ratio. The residues obtained were screened for their cytotoxic activity by MTT assay in HeLa and MCF-7 cell lines. The ethanol extract of F. hispida (Moraceae) and A. scholaris (Apocyanaceae) leaves exhibited excellent cytotoxic activity with an IC50 value of 8.47µg/mL in HeLa and 10.23 µg/mL in MCF-7 cell lines, respectively. The alcoholic leaf extract of A. sholaris showed IC50 values of 20.62 µg/mL and 18.50 µg/mL in HeLa and MCF-7 cells respectively.

Sunil DHANYA; N. V. Anil KUMAR; Anand Shivaram NAYAK; U. Sachin RAJ; Deepa PRABHU; RAVIKIRAN

2013-01-01

309

The induction of DNA synthesis in the chick red cell nucleus in heterokaryons during the first cell cycle after fusion with HeLa cells.  

UK PubMed Central (United Kingdom)

Induction of DNA synthesis in embryonic chick red cells has been examined during the first and second cell cycles after fusion with HeLa cells synchronized in different parts of G1 and S-phase. The data indicate that: (i) the younger the embryonic blood the more rapidly the red cells are induced into DNA synthesis; (ii) the greater the ratio of HeLa to chick nuclei in the heterokaryon, the more rapidly the induction occurs; (iii) DNA synthesis in the chick nucleus can continue after the HeLa nucleus has left S-phase and entered either G2 or mitosis; (iv) the induction potential of late S-phase HeLa is somewhat lower than that of early or mid S-phase cells; (v) less than 10% of the chick DNA is replicated during the first cycle after fusion and only a small proportion (15%) of the chick nuclei approach the 4C value of DNA during the second cycle after fusion; (vi) the newly synthesized DNA is associated either with the condensed regions of the nucleus or with the boundaries between condensed and non-condensed regions; (vii) the chick chromosomes at the first and second mitosis after fusion are in the form of PCC prematurely condensed chromosomes); they are never fully replicated and are often fragmentary; (viii) DNA synthesis in the chick nuclei is accompanied by an influx of protein (both G1 and S-phase protein) from the HeLa component of the heterokaryon.

Johnson RT; Mullinger AM

1975-08-01

310

Dose-rate effects in plateau-phase cultures of S3 HeLa and V79 cells  

International Nuclear Information System (INIS)

[en] Dose-rate effects on cell survival were studied for log-, fed plateau-, and unfed plateau-phase cultures of V79 and S3 HeLa cells. For log-phase cultures, repair, cell-cycle redistribution, and cell division during exposure can contribute to the overall dose-rate effect, but their relative contributions are difficult to determine. With plateau-phase cultures, the cell-cycle times are greatly lengthened, for those cells that are in cycle. Hence, the contribution to the overall dose-rate effect of cell-cycle redistribution and cell division during the exposure could be minimized using plateau-phase cultures. With respect to the acute dose-survival curves, there was a clear loss in effectiveness when the dose rate was lowered to 154 rad/hr for both fed and unfed plateau-phase HeLa and V79 cells. There was no further reduction in effectiveness per unit dose, however, when the dose rate was reduced to 55 rad/hr. Since there was virtually no cell division or cell-cycle redistribution, it may be that a limit to the repair-dependent dose-rate effect at 370C has been reached at a dose rate of 154 rad/hr

1979-01-01

311

Uptake of (/sup 3/H)ouabain from the cell surface into the lysosomal compartment of HeLa cells  

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(/sup 3/H)Ouabain specifically bound at sublethal concentrations to Na,K-ATPase on the surface of HeLa cells is taken up (internalized) by the cells at a rate of three membrane equivalents of labeled sites per generation. Immediately following a pulse label with the glycoside, codistribution of radioactivity with the surface marker 5'-nucleotidase is found in both conventional sucrose-gradient fractionation and in fractionation following a digitonin treatment. At appropriate concentrations digitonin increases the buoyant density of the HeLa surface membrane and solubilizes the lysosomal marker ..beta..-hexosaminidase (Tulkens et al., 1974). After internalization, (/sup 3/H)ouabain is also solubilized by digitonin. A shear analysis is described which shows internalized ouabain and ..beta..-hexosaminidase to be codistributed in a particulate fraction that is homogeneous with respect to shear; extrapolation to zero-shear shows that little or none of either marker is found in the soluble fraction of the cytosol. Both markers are coreleased from the particulate fraction by osmotic shock. Although internalized ouabain is subsequently released from these cells with a half-time of about 70 hr, apparently by exocytosis, the shear sensitivity of the remaining cell-associated ouabain does not change for up to 72 hr. Thus ouabain (together with Na,K-ATPase.) appears to be taken up from the surface into a lysosomal compartment and, by at least one criterion, this compartment does not change its physical properties with time, i.e., does not ''age.''

Cook, J.S.; Tate, E.H.; Shaffer, C.

1982-02-01

312

Observations of the first postirradiation division of HeLa cells following continuous or fractionated exposure to ? rays  

International Nuclear Information System (INIS)

[en] The first postirradiation division of synchronized S3 HeLa cells was studied using both continuous and fractionated irradiation treatments. Synchronized HeLa cells continuously irradiated at a dose rate of 37 rad/hr eventually accumulate in mitosis. If the continuous irradiation is stopped before the cells enter G2 or even after they have progressed for a limited time into the G2 arrest that develops, very little subsequent accumulation of cells in mitosis occurs. If they progress for a longer time into the G2 arrest, then some mitotic accumulation does occur after the irradiation is stopped. When synchronized cells were allowed to progress through G1 and S before the irradiation was started, very little cell division occurred during subsequent continuous irradiation and extensive mitotic accumulation was observed. Thus, for continuous irradiation of HeLa cells, the dose received by a cell during G2 or a G2 delay apparently determines whether it will be able to divide if it reaches mitosis. Arguing against the notion that continuous irradiation during G2 is required to produce a mitotic accumulation was the result of an expriment which showed that a similar effect was obtained using two acute doses: the first to produce a G2 delay and the second to give the necessary dose during the delay. The first dose alone resulted in little mitotic accumulation. The time of delivery of the second dose during the G2 delay affected the extent of mitotic accumulation observed. There was less mitotic accumulation when second acute doses were given early or at intermediate times during the delay than when they were given late during the G2 delay. An accumulation of cells in mitosis was also observed by using a combination of low-dose-rate irradiation to induce a G2 delay, followed immediately by an acute dose of either 500 or 1000 rad. The low-dose-rate treatment alone resulted in no mitotic accumulation

1979-01-01

313

Effect of combined treatment of HeLa S3 cells with radiation and etoposide on cell survival  

International Nuclear Information System (INIS)

[en] Etoposide, a semisynthetic derivative of podophyllotoxin, is an active cytotoxic agent. In this paper, radiation sensitivity of HeLa S3 cells was evaluated with and without etoposide. Exponentially growing monolayer cells were incubated with etoposide at 2.5 ?g/ml for 1 hour immediately after irradiation. The combination of radiation and etoposide showed a significant enhancement in cell killing as compared with radiation alone. The difference in surviving fraction is principally due to the decrease of the shoulder of the cell survival curve. When etoposide was combined with radiation at the different time intervals in exponentially growing cells, a significant enhancement of cell killing was found immediately before and after radiation, and at 6 hours after radiation. When plateau phase cells were incubated with etoposide at 2.5 ?g/ml for 1 hour immediately after radiation, no enhancement of cell killing was observed. These results are discussed in connection with the cellular repair and cell cycle kinetics. (author)

1988-01-01

314

Effect of combined treatment of HeLa S3 cells with radiation and etoposide on cell survival  

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Etoposide, a semisynthetic derivative of podophyllotoxin, is an active cytotoxic agent. In this paper, radiation sensitivity of HeLa S3 cells was evaluated with and without etoposide. Exponentially growing monolayer cells were incubated with etoposide at 2.5 ..mu..g/ml for 1 hour immediately after irradiation. The combination of radiation and etoposide showed a significant enhancement in cell killing as compared with radiation alone. The difference in surviving fraction is principally due to the decrease of the shoulder of the cell survival curve. When etoposide was combined with radiation at the different time intervals in exponentially growing cells, a significant enhancement of cell killing was found immediately before and after radiation, and at 6 hours after radiation. When plateau phase cells were incubated with etoposide at 2.5 ..mu..g/ml for 1 hour immediately after radiation, no enhancement of cell killing was observed. These results are discussed in connection with the cellular repair and cell cycle kinetics.

Kubota, Nobuo; Ikegami, Tadashi; Watai, Kiichi; Kakehi, Masae; Matsui, Kengo

1988-10-01

315

Dihydroartemisinin induces radiosensitivity in cervical cancer cells by modulating cell cycle progression.  

UK PubMed Central (United Kingdom)

OBJECTIVE: To investigate the radiosensitizing effects of dihydroartemisinin (DHA) and its underlying mechanisms in cervical cancer cells. METHODS: This experimental study was conducted between May 2009 and August 2012 in the School of Radiation Medicine and Protection, Soochow University, Suzhou, China. HeLa and Siha cells were assigned as the control group and DHA as treated group. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, clonogenic assay, cell cycle analysis, and apoptosis analysis were carried out in 2 cell lines of both groups. RESULTS: The inhibitory effect of DHA on the HeLa and Siha cell lines was dependent on both concentration and time. Dihydroartemisinin increased the radiosensitivity of HeLa cells, but not of Siha cells. Apoptosis and the gap2/mitosis (G2/M) phase transition induced by x-irradiation was enhanced by DHA treatment in HeLa cells. Irradiation, combined with DHA, decreased Wee1 expression while increasing Cyclin B1 expression in HeLa cells. CONCLUSION: Dihydroartemisinin potently abrogates G2 checkpoint control in HeLa cells. It can relieve the G2/M arrest induced by irradiation; thus, it can be used as an effective radiosensitizer, which will probably promote the entry of more irradiation-damaged cells into mitosis.

Luo J; Chen X; Chen G; Zhou X; Lu X; Ling Y; Zhang S; Zhu W; Cao J

2013-03-01

316

Dihydroartemisinin induces radiosensitivity in cervical cancer cells by modulating cell cycle progression  

Directory of Open Access Journals (Sweden)

Full Text Available Objectives: To investigate the radiosensitizing effects of dihydroartemisinin (DHA) and its underlying mechanisms in cervical cancer cells. Methods: This experimental study was conducted between May 2009 and August 2012 in the School of Radiation Medicine and Protection, Soochow University, Suzhou, China. HeLa and Siha cells were assigned as the control group and DHA as treated group. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, clonogenic assay, cell cycle analysis, and apoptosis analysis were carried out in 2 cell lines of both groups. Results: The inhibitory effect of DHA on the HeLa and Siha cell lines was dependent on both concentration and time. Dihydroartemisinin increased the radiosensitivity of HeLa cells, but not of Siha cells. Apoptosis and the gap2/mitosis (G2/M) phase transition induced by x-irradiation was enhanced by DHA treatment in HeLa cells. Irradiation, combined with DHA, decreased Wee1 expression while increasing Cyclin B1 expression in HeLa cells. Conclusion: Dihydroartemisinin potently abrogates G2 checkpoint control in HeLa cells. It can relieve the G2/M arrest induced by irradiation; thus, it can be used as an effective radiosensitizer, which will probably promote the entry of more irradiation-damaged cells into mitosis. 

Judong Luo; Xialin Chen; Guanglie Chen; Xifa Zhou; Xujing Lu; Yang Ling; Shuyu Zhang; Wei Zhu; Jianping Cao

2013-01-01

317

Analysis of gene expression profiles in HeLa cells in response to overexpression or siRNA-mediated depletion of NASP  

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Full Text Available Abstract Background NASP (Nuclear Autoantigenic Sperm Protein) is a linker histone chaperone required for normal cell division. Changes in NASP expression significantly affect cell growth and development; loss of gene function results in embryonic lethality. However, the mechanism by which NASP exerts its effects in the cell cycle is not understood. To understand the pathways and networks that may involve NASP function, we evaluated gene expression in HeLa cells in which NASP was either overexpressed or depleted by siRNA. Methods Total RNA from HeLa cells overexpressing NASP or depleted of NASP by siRNA treatment was converted to cRNA with incorporation of Cy5-CTP (experimental samples), or Cy3-CTP (control samples). The labeled cRNA samples were hybridized to whole human genome microarrays (Agilent Technologies, Wilmington, Delaware, USA). Various gene expression analysis techniques were employed: Significance Analysis of Microarrays (SAM), Expression Analysis Systematic Explorer (EASE), and Ingenuity Pathways Analysis (IPA). Results From approximately 36 thousand genes present in a total human genome microarray, we identified a set of 47 up-regulated and 7 down-regulated genes as a result of NASP overexpression. Similarly we identified a set of 56 up-regulated and 71 down-regulated genes as a result of NASP siRNA treatment. Gene ontology, molecular network and canonical pathway analysis of NASP overexpression demonstrated that the most significant changes were in proteins participating in organismal injury, immune response, and cellular growth and cancer pathways (major "hubs": TNF, FOS, EGR1, NF?B, IRF7, STAT1, IL6). Depletion of NASP elicited the changed expression of proteins involved in DNA replication, repair and development, followed by reproductive system disease, and cancer and cell cycle pathways (major "hubs": E2F8, TP53, FGF, FSH, FST, hCG, NF?B, TRAF6). Conclusion This study has demonstrated that NASP belongs to a network of genes and gene functions that are critical for cell survival. We have confirmed the previously reported interactions between NASP and HSP90, HSP70, histone H1, histone H3, and TRAF6. Overexpression and depletion of NASP identified overlapping networks that included TNF as a core protein, confirming that both high and low levels of NASP are detrimental to cell cycle progression. Networks with cancer-related functions had the highest significance, however reproductive networks containing follistatin and FSH were also significantly affected, which confirmed NASP's important role in reproductive tissues. This study revealed that, despite some overlap, each response was associated with a unique gene signature and placed NASP in important cell regulatory networks.

Alekseev Oleg M; Richardson Richard T; Alekseev Oleg; O'Rand Michael G

2009-01-01

318

Cell killing and division delay in asynchronous and synchronized HeLa cells irradiated with alpha particles or x rays  

International Nuclear Information System (INIS)

HeLa cells irradiated with a single or two split doses of ? particles or X rays were observed with time-lapse photography or examined for their colony-forming ability. The cell cycle-dependent variation of cell killing and division delay were compared in synchronous and asynchronous cell populations. Cellular damage by ? particles was manifested in the form of cessation of division, or death, rather than partial division which was predominant for X irradiation. The pattern of cell killing with ? particles was similar to that found with X rays, in that high sensitivity was noted at or close to mitosis, while a resistant peak remained at late S but not in early G1. The pattern of division delay was similar for X rays and ? particles during G2-M, with a maximum delay at mid G2 and no delay past the transition point, but differed during G1-S. During this period, division delay increased with cell age, whereas it showed a broad peak at G1-S boundary and a trough at late S for X rays. However, such was not the case for ? particles.

1984-01-01

319

Synthesis and methylation of ribosomal RNA in HeLa cells infected with the herpes virus pseudorabies virus  

Energy Technology Data Exchange (ETDEWEB)

The effects of infection with the herpes virus pseudorabies virus on the metabolism of HeLa cell ribosomal RNA were examined. There is a decline both in the synthesis of nucleolar 45S ribosomal precursor RNA and in its processing to mature cytoplasmic RNA. The methylated oligonucleotides in the ribosomal RNA species were studied. The methylation of cytoplasmic ribosomal RNA was essentially unchanged. However there was some undermethylation of the nucleolar precursor. If undermethylated RNA does not mature then this may partly explain the reduced processing in the infected cells.

Furlong, J.C.; Kyriakidis, S.; Stevely, W.S. (Glasgow Univ. (UK))

1982-01-01

320

Synthesis and methylation of ribosomal RNA in HeLa cells infected with the herpes virus pseudorabies virus  

International Nuclear Information System (INIS)

[en] The effects of infection with the herpes virus pseudorabies virus on the metabolism of HeLa cell ribosomal RNA were examined. There is a decline both in the synthesis of nucleolar 45S ribosomal precursor RNA and in its processing to mature cytoplasmic RNA. The methylated oligonucleotides in the ribosomal RNA species were studied. The methylation of cytoplasmic ribosomal RNA was essentially unchanged. However there was some undermethylation of the nucleolar precursor. If undermethylated RNA does not mature then this may partly explain the reduced processing in the infected cells. (Author)

1982-01-01

 
 
 
 
321

Pronounced transcriptional regulation of apoptotic and TNF-NF-kappa-B signaling genes during the course of thymoquinone mediated apoptosis in HeLa cells.  

UK PubMed Central (United Kingdom)

Thymoquinone (TQ) is the active ingredient extracted from the essential oil of Nigella sativa. A number of studies implicated TQ as an antitumor agent. In this study, cytotoxic effects of the oil of N. sativa and TQ were evaluated on human cervical cancer cell line, HeLa cells. IC50 value was ~0.125 ?l/ml for N. sativa oil preparations and 12.5 ?M for TQ. TQ strongly inhibited wound healing at all concentrations ranging from 12.5 to 100 ?M in a scratch wound healing assay. Additionally, induction of apoptosis by TQ was assessed by Giemsa staining and TQ was found to induce apoptosis in cancer cells especially at concentrations of 50 and 100 ?M. TQ-mediated transcriptional regulation of 84 genes involved in apoptosis was studied using a PCR array. At low dose (12.5 ?M), TQ was found to induce expression of four pro-apoptotic genes: BIK (~22.7-fold), FASL (~2.9-fold), BCL2L10 (~2.1-fold), and CASP1 (~2-fold). TQ was also found to reduce the expression of an anti-apoptotic gene implicated in NF-kappa-B signaling and cancer: RELA (~8-fold). At high dose (100 ?M), TQ mediated the expression of 21 genes implicated directly in apoptosis (6 genes), TNF signaling (10 genes), and NF-kappa-B signaling (3 genes) such as BIK, BID, TNFRSF10A, TNFRSF10B, TNF, TRAF3, RELA, and RELB. In conclusion, this study implicates the role of TQ in the inhibition of cancer cell proliferation and migration. At the same time, our results strongly suggest that TQ intervenes with TNF and NF-kappa-B signaling during TQ-mediated induction of apoptosis in cancer cells.

Sakalar C; Yuruk M; Kaya T; Aytekin M; Kuk S; Canatan H

2013-08-01

322

Induction of apoptosis in human cervical carcinoma HeLa cells by polymethoxylated flavone-rich Citrus grandis Osbeck (Dangyuja) leaf extract.  

Science.gov (United States)

Citrus grandis Osbeck (Dangyuja) has a high content of flavonoids with health-related properties. Although previous data have revealed the anticancer potency of some Citrus species, the underlying molecular mechanisms of this activity by leaf extracts have not been studied in detail. The purpose of this study was to evaluate the cytotoxic effects of citrus leaves on five human cancer cell lines and to determine the possible mechanisms of cell death elicited by the chloroform fraction (CF) of the Dangyuja leaf. The CF of Dangyuja strongly decreased the survival rate of HeLa cells, among the tested cell lines. CF treatment induced the down-regulation of anti-apoptotic Bcl-2 expression, resulting in the proteolytic activation of caspases and the degradation of poly (ADP-ribose) polymerase (PARP) protein. Arrested cell growth and induction of apoptosis were confirmed by flow cytometry and DNA fragmentation analysis, respectively. The major components of the CF were identified as isosinensetin, sinensetin, tetramethyl-O-isoscutellarein, nobiletin, tangeretin, and 5-hydroxy-6,7,8,3',4'-pentamethoxyflavone by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). Our results suggest that the CF of Dangyuja leaves is an excellent source of functional polymethoxylated flavones, which may help prevent cervical cancer and may potentially be a useful agent for the treatment of certain malignancies. PMID:20538032

Kim, Hana; Moon, Jeong Yong; Mosaddik, Ashik; Cho, Somi Kim

2010-06-09

323

Relationship of ROS and NO in X-ray induced bystander effects in HeLa cells  

International Nuclear Information System (INIS)

[en] Accumulating evidence indicates that irradiated cells can release signals which induce a series of biological responses in non-exposed cells. This is known as irradiation-induced bystander effects. Both reactive oxygen species(ROS)and nitric oxide(NO) play important roles in bystander effects. In this study, we determined the relationship of ROS and NO in the signaling pathway of bystander effects. HeLa cells were treated with or without dimethy sulfoxide(DMSO) before X-ray irradiation, and micronuclei formation as well as cell proliferation rate was detected in both irradiated and bystander cells. In addition,we also detected inducible nitric oxide synthase(iNOS)expression and NO level in irradiated cells using Western blotting and DAF-FM DA fluorescent probe, respectively. Our results showed that micronuclei were induced in irradiated and bystander cells while DMSO treatment significantly suppressed the formation of micronuclei in both of them. We also found that when cells were irradiated their proliferation rate was suppressed while DMSO treatment eliminated this inhibition effect.In contrast, the cells received conditioned medium from irradiated cells proliferated more quickly than the cells received medium from non-irradiated cells while DMSO treatment reduced the difference. Finally, we found that irradiated cells had higher level of iNOS and NO compared to non-irradiated controls, whereas DMSO treatment decreased their levels. These results suggest that ROS is the upstream signal of NO in X-ray induced bystander effects in HeLa cells. (authors)

2009-01-01

324

[Detection of cytotoxicity of biocompatible materials in MRC-5, HeLa, and RC-IAL cell lines  

UK PubMed Central (United Kingdom)

The sensitivity of diploid and heteroploid cell lines for detection of cytotoxicity using the agar diffusion method on cell culture, was tested with ascorbic acid solution of different concentrations. A total of 562 samples of 21 various materials were tested. The heteroploid cell line, RC-IAL, showed in relation to the MRC-5 and HeLa cell lines, greater sensitivity because it showed the presence of cytotoxic effect with the lowest concentration used (10 and 25 micrograms/ml) of ascorbic acid and showed greater diameter of cytotoxic halo in 15 samples and equal diameter in 16 of the 43 positive samples (7.6%). Out of 43 positive samples, the MRC-5 line did not show cytotoxicity in 3 sponge samples and 1 of acrylic resin. The PVC (polyvinylchloride) and polyethylene rarely showed positivity, while, the plastic, cotton and acrylic resin demonstrated cytotoxicity in about 5% of samples. We thus suggest the use of the RC-IAL and HeLa cell lines for continuation of this type of analysis at Adolfo Lutz Institute.

Cruz AS; Figueiredo CA; Martinez CH; Gomes LF

1992-03-01

325

Sulfated fucan from marine alga inhibits HeLa cells infection by HTLV-1 free particles: semi-quantitative analysis  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english A sulfated fucan from Laminaria abyssalis marine alga prevented the interaction of HTLV-1 particles, purified from the MT-2 cell line, with HeLa cells. The infection obtained using a concentrated virus suspension was detected only by amplification of the newly synthesized HTLV-1 proviral cDNA by the nested-polymerase chain reaction (PCR). The sulfated polysaccharide was not toxic to the cells at a concentration of 100 µg/mL and prevented infection by the viral particles (more) when added to the cell monolayers. The proviral cDNA was only detected when the sulfated polysaccharide was added to the cells three hours post-infection, indicating that the inhibitory activity occurred in the initial stages of virus-cell interaction. Our results demonstrate, for the first time, the ability of a sulfated fucan from marine algae to inhibit virus transmission through free virus particles.

Romanos, Maria T. V.; Andrada-Serpa, Maria J.; Mourão, Paulo A. S.; Yoneshigue-Valentin, Yocie; Pereira, Mariana S.; Santos, Norma; Wigg, Marcia D.

2011-04-01

326

Lactoferrin and free secretory component of human milk inhibit the adhesion of enteropathogenic Escherichia coli to HeLa cells  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Diarrhoea caused by Escherichia coli is an important cause of infant morbidity and mortality in developing countries. Enteropathogenic Escherichia coli (EPEC) is considered one of the major causes of diarrhoea in children living in developing countries. The ability of diarrhoeagenic strains of E. coli to adhere to and colonize the intestine is the first step towards developing the disease. EPEC strains adhere to enterocytes and HeLa cells in a characteristic pattern known as localized adherence. Many epidemiological studies of diarrhoea have shown that breast-feeding protects infants from intestinal infections. Both immunoglobulin and non-immunoglobulin elements of human milk are thought to contribute to the protection from diarrhoeal agents. Results The effects of human milk and its protein components on the localized adherence of EPEC were investigated. Non-immunoglobulin components of human milk responsible for the inhibition of EPEC adhesion to HeLa cells were isolated by chromatographic fractionation of human whey proteins. Besides secretory immunoglobulin A, which has been previously reported to affect the adhesion of EPEC, free secretory component (fSC) and lactoferrin (Lf) were isolated. Even in concentrations lower than those usually found in whole milk, fSC and Lf were able to inhibit the adhesion of EPEC. ?-lactalbumin was also isolated, but showed no activity on EPEC adhesion. Conclusions This study demonstrated that the immunoglobulin fraction, the free secretory component and lactoferrin of human milk inhibit EPEC adhesion to HeLa cells. These results indicate that fSC and Lf may be important non-specific defence factors against EPEC infections.

de Araújo Andréa; Giugliano Loreny

2001-01-01

327

Effects of DNA polymerase inhibitors on replicative and repair DNA synthesis in ultraviolet-irradiated HeLa cells  

Energy Technology Data Exchange (ETDEWEB)

Aphidicolin specifically inhibits eukaryotic DNA polymerase ..cap alpha.., while 2',3'-dideoxythymidine 5'-triphosphate (d/sub 2/TTP) inhibits DNA polymerase ..beta.. and ..gamma.. but not ..cap alpha... 1-..beta..-D-Arabinofuranosylcytosine 5'-triphosphate (araCTP) inhibits both DNA polymerase ..cap alpha.. and ..beta.. although to a different extent. Here we measured the effects of these inhibitors on repair DNA synthesis of U.V.-irradiated HeLa cells by two different methods. Firstly, aphidicolin, 1-..beta..-D-arabinofuranosylcytosine (araC, a precursor of araCTP) and 2',3'-dideoxythimidine (d/sub 2/Thd, a precursor of d/sub 2/TTP) were added directly to the culture medium. In this case, aphidicolin and araC strongly inhibited replicative DNA synthesis of HeLa cells, and they also inhibited repair synthesis after U.V.-irradiation but to a much lesser extent. In contrast, high concentrations of d/sub 2/Thd inhibited repair DNA synthesis to a higher extent than replicative DNA synthesis. Secondly, the active form of inhibitor, d/sub 2/TTP, was microinjected directly into cytoplasm or nuclei or U.V.-irradiated HeLa cells. Microinjection of d/sub 2/TTP effectively inhibited repair synthesis. The microinjection of d/sub 2/TTP, into either cytoplasm or nucleus, strongly inhibited replicative synthesis. These results might indicate that multiple DNA polymerases are involved in repair synthesis as well as in replicative synthesis.

Morita, T.; Nakamura, H. (Aichi Cancer Center, Nagoya (Japan)); Tsutsui, Y.; Nishiyama, Y. (Nagoya Univ. (Japan). Faculty of Medicine); Yoshida, S. (Aichi Prefectural Colony, Kasugai (Japan). Inst. for Developmental Research)

1982-11-01

328

Extended Electrical Model for Impedance Characterization of Cultured HeLa Cells in Non-Confluent State Using ECIS Electrodes.  

UK PubMed Central (United Kingdom)

Electric cell substrate impedance sensing has been widely used as a label free quantitative platform to study various cell biological processes and it is extremely essential to extract the parameters like the variation of the cell substrate spacing, changing projected area of the cell on the electrode and approximate cluster size during the non-confluent state to understand the mechanism of proliferation of the cells. The distributed analytical models developed so far to extract these parameters are applicable only under the conditions when the cells have become confluent. There are some lumped electrical models which have been reported for the non-confluent state but they do not provide correct estimate of the changing cell substrate spacing and the cell cluster size during growth. In this paper we develop extended distributed electrical models to characterize the impedance spectroscopy behavior of cultured HeLa cells in 200 Hz to 1 MHz range using eight well ECIS electrodes in the non-confluent state. The distributed model introduces some pseudo regularity in the arrangement of the non-confluent cells to extract the average ensemble of the significant parameters. The parameters extracted from the distributed model after 10 hours, 20 hours, and 30 hours of HeLa cell growth have been compared with the lumped circuit model and has been observed to fit the experimental data with a seven times improved fit quality factor. Further, the changing cell radius and cluster radius extracted at three different instants of time from the distributed analytical model have been found to match closely the microscopic observation in contrast to the lumped circuit model.

Mondal D; Roychaudhuri C

2013-09-01

329

Inhibition of the MRP1-mediated transport of the menadione-glutathione conjugate (thiodione) in HeLa cells as studied by SECM  

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Oxidative stress induced in live HeLa cells by menadione (2-methyl-1,4-napthaquinone) was studied in real time by scanning electrochemical microscopy (SECM). The hydrophobic molecule menadione diffuses through a living cell membrane where it is toxic to the cell. However, in the cell it is conjugate...

Koley, Dipankar; Bard, Allen J.

330

The fibrate decreases radiation sensitivity via peroxisome proliferator-activated receptor {alpha}-mediated superoxide dismutase induction in HeLa cells  

Energy Technology Data Exchange (ETDEWEB)

The fibrates are ligands for peroxisome proliferator-activated receptor (PPAR) {alpha} and used clinically as hypolipidemic drugs. The fibrates are known to cause peroxisome proliferation, enhance superoxide dismutase (SOD) expression and catalase activity. The antioxidant actions of the fibrates may modify radiation sensitivity. Here, we investigated the change of the radiation sensitivity in two cervix cancer cell lines in combination with fenofi brate (FF). Activity and protein expression of SOD were measured according to the concentration of FF. The mRNA expressions were measured by using real time reverse-transcription polymerase chain reaction. Combined cytotoxic effect of FF and radiation was measured by using clonogenic assay. In HeLa cells total SOD activity was increased with increasing FF doses up to 30 {mu}M. In the other hand, the catalase activity was increased a little. As with activity the protein expression of SOD1 and SOD2 was increased with increasing doses of FF. The mRNAs of SOD1, SOD2, PPAR{alpha} and PPAR{gamma} were increased with increasing doses of FF. The reactive oxygen species (ROS) produced by radiation was decreased by preincubation with FF. The surviving fractions (SF) by combining FF and radiation was higher than those of radiation alone. In Me180 cells SOD and catalase activity were not increased with FF. Also, the mRNAs of SOD1, SOD2, and PPAR{alpha} were not increased with FF. However, the mRNA of PPAR{gamma} was increased with FF. FF can reduce radiation sensitivity by ROS scavenging via SOD induction in HeLa. SOD induction by FF is related with PPAR{alpha}.

Liu, Xianguang; An, Zhengzhe; Song, Hye Jin; Kim, Won Dong; Park, Woo Yoon [Chungbuk National University College of Medicine, Cheongju (Korea, Republic of); Jang, Seong Soon [The Catholic University of Korea College of Medicine, Seoul (Korea, Republic of); Yu, Jae Ran [Konkuk University College of Medicine, Chungju (Korea, Republic of)

2012-06-15

331

Influence of He-Ne laser radiation on HeLa cell survival rate after gamma-irradiation  

International Nuclear Information System (INIS)

A monolayer of HeLa cells, at the stationary phase of growth, exposed to He-Ne laser radiation either 5 min or 60 min prior to gamma-irradiation (0.1-10 Gy; 6.75 Gy/min), or 5 min after irradiation has been investigated. With a 5-min interval between irradiation sessions (both sequences) the survival curves are virtually the same as those for gamma-irradiated cells only. With He-Ne laser radiation delivered 60 min before gamma-irradiation with doses exceeding 5 Gy, fraction of radioresistant cells is identified whose D0 is almost twice as thigh as D0 basic cell mass. A hypothesis is proposed that He-Ne laser the radiation activates, in some cells, the processes that promote the repair of radiation damages.

1992-01-01

332

Lung cancer - small cell  

Science.gov (United States)

Cancer - lung - small cell; Small cell lung cancer; SCLC ... About 15% of all lung cancer cases are SCLC. Small cell lung cancer is slightly more common in men than women. Almost all cases of SCLC ...

333

In vitro uptake and release of 201Tl and 99mTc-MIBI in HeLa cell  

International Nuclear Information System (INIS)

201Tl is useful in the diagnosis of tumor malignancy determined by the grade of washout rate to the normal tissue especially in lung tumors and thyroid tumors. 99mTc-MIBI, a tracer of myocardial blood flow is also a tumor tracer. We determined whether the growth of the tumor cell is related to the uptake and release of these tracers in cultured cells (HeLa cell). Cultured HeLa cells were incubated for 1 hr with 10 kBq of either tracer for the kinetic study of cellular uptake and additionally incubated for 90 min with cold medium for the kinetic study of cellular release. These cells cultured with various concentrations of actinomycin D (ACD) were used to examine the correlation between cellular growth and kinetics of these tracers. The uptake in the cell reached a plateau at about 60 min and the uptake at 60 min correlated with the number of the cells and was identified for specific accumulation in the cell. The uptake of 201Tl and 99mTc-MIBI at 60 min was 5.17% and 1.32% respectively of the given dose. The release for 90 min showed almost the same tendency for these tracers. The cellular release of 201Tl with the addition of ACD was increased in association with the concentration of ACD. On the other hand, the cellular release of 99mTc-MIBI showed no change by the addition of ACD. In conclusion, 201Tl showed slower washout in high growth cells than in low growth cells, and could act as an indicator of tumor malignancy by assessment of its washout by the tumor. (author)

1995-01-01

334

Size-dependent cellular uptake efficiency, mechanism, and cytotoxicity of silica nanoparticles toward HeLa cells.  

UK PubMed Central (United Kingdom)

In this study, we investigated and reported the cellular uptake efficiency, mechanism, and cytotoxicity of silica nanoparticles (SNPs) with different sizes. Using confocal laser scanning microscope (CLSM), flow cytometry (FCM), and graphite furnace atomic absorption spectrometry (GFAAS), the qualitative and quantitative experimental results showed that the cellular uptake of SNPs toward HeLa cells is size-dependent. To further examine the uptake process, three different inhibitors including sucrose, Filipin III, and Cytochalasin D (Cyt D) were introduced to pretreat the HeLa cells. It appeared that the largest SNPs (SNPs-307.6) take an energy-dependent uptake pathway (clathrin dependent and caveolin independent) while that for the medium size SNPs-167.8 involves clathrin and caveolin dependent endocytosis. In contrast, the smallest SNPs (SNPs-55.6) follow not only energy required clathrin and caveolin dependent endocytosis but also an energy independent pathway to efficiently enter the cells. Moreover, the cellular uptake efficiency of SNPs, which also show excellent biocompatibility, is size-dependent in the order of 55.6>167.8>307.6 nm. This knowledge is fundamentally important and will facilitate more development of size-defined SNPs as the transporters for various purposes.

Zhu J; Liao L; Zhu L; Zhang P; Guo K; Kong J; Ji C; Liu B

2013-03-01

335

Radioadaptive response to the medium-mediated bystander induction of DNA strand breaks in HeLa cells  

International Nuclear Information System (INIS)

[en] Full text: Numerous investigators have reported two cellular responses of importance at low doses that have a potential impact on the risk estimation of ionizing radiation. The radioadaptive response confers resistance to a subsequent dose by a low priming dose, while the bystander effect exaggerates the effect of small doses. The present study was conducted to examine the interaction of the radioadaptive response with the bystander effect in HeLa cells. The culture was irradiated with 0.5 to 8 Gy of 140 kVp X-rays and one hour later, the medium was taken, passed through a filter and transferred to the parallel culture of non-irradiated HeLa cells as non-targeted cells. After incubation for 30 min, the induced DNA damage was analyzed by the single cell gel-electrophoresis assay under alkaline or neutral conditions. The treatments resulted in a dose-dependent increase in tail moment under either conditions, indicating the induction of DNA single- and double-strand breaks. The clonogenic survival of non-irradiated cells was also reduced after they were cultured in the medium that was taken from irradiated cultures. Any change was not observed when the medium alone was irradiated. These results give the disputed evidence that certain genotoxic factor(s) released from irradiated cells into the culture medium can induce DNA strand breaks leading to cell death. It is also suggested that physical contact between irradiated and non-irradiated cells may not be required for the bystander effect. In adapted cells that were pre-exposed to 5 cGy of X-rays and cultured for 4 h beforehand, the yield of DNA strand breaks induced by X-rayed medium was reduced by about 50 %. The results, in conjunction with our early finding (Ikushima et al., 1996) suggest that the radioadaptive response resulting from such a low dose may diminish the bystander effect through an enhanced DNA repair function

2003-01-01

336

Using a micro electroporation chip to determine the optimal physical parameters in the uptake of biomolecules in HeLa cells.  

UK PubMed Central (United Kingdom)

In this study, a new micro electroporation (EP) cell chip with three-dimensional (3D) electrodes was fabricated by means of MEMS technology, and tested on cervical cancer (HeLa) cells. Extensive statistical data of the threshold electric field and pulse duration were determined to construct an EP "phase diagram", which delineates the boundaries for 1) effective EP of five different size molecules and 2) electric cell lysis at the single-cell level. In addition, these boundary curves (i.e., electric field versus pulse duration) were fitted successfully with an exponential function with three constants. We found that, when the molecular size increases, the corresponding electroporation boundary becomes closer to the electric cell lysis boundary. Based on more than 2000 single-cell measurements on five different size molecules, the critical size of molecule was found to be approximately 40 kDa. Comparing to the traditional instrument, MEMS-based micro electroporation chip can greatly shorten the experimental time.

He H; Chang DC; Lee YK

2007-05-01

337

Using a micro electroporation chip to determine the optimal physical parameters in the uptake of biomolecules in HeLa cells.  

Science.gov (United States)

In this study, a new micro electroporation (EP) cell chip with three-dimensional (3D) electrodes was fabricated by means of MEMS technology, and tested on cervical cancer (HeLa) cells. Extensive statistical data of the threshold electric field and pulse duration were determined to construct an EP "phase diagram", which delineates the boundaries for 1) effective EP of five different size molecules and 2) electric cell lysis at the single-cell level. In addition, these boundary curves (i.e., electric field versus pulse duration) were fitted successfully with an exponential function with three constants. We found that, when the molecular size increases, the corresponding electroporation boundary becomes closer to the electric cell lysis boundary. Based on more than 2000 single-cell measurements on five different size molecules, the critical size of molecule was found to be approximately 40 kDa. Comparing to the traditional instrument, MEMS-based micro electroporation chip can greatly shorten the experimental time. PMID:16820330

He, Huiqi; Chang, Donald C; Lee, Yi-Kuen

2006-05-23

338

Yatein from Chamaecyparis obtusa suppresses herpes simplex virus type 1 replication in HeLa cells by interruption the immediate-early gene expression.  

Science.gov (United States)

Inhibitory effects of methanolic extracts from nine Chinese herbs on herpes simplex virus type 1 (HSV-1) replication were studied. By a bioassay-guided fractionation procedure, yatein (C(22)H(23)O(7); M.W.399) was isolated from Chamaecyparis obtusa; yatein significantly suppressed HSV-1 multiplication in HeLa cells without apparent cytotoxicity. To further localize the point in the HSV-1 replication cycle where arrest occurred, a set of key regulatory events leading to the viral multiplication was examined, including viral immediate-early (alpha) and late (gamma) gene expression and DNA replication. Results indicated that levels of glycoprotein B (gB) and gC mRNA expression in HeLa cells were impeded by yatein. Data from polymerase chain reaction showed that replication of HSV-1 DNA in HeLa cells was arrested by yatein. Furthermore, yatein decreased ICP0 and ICP4 gene expression in HeLa cells. Results of an electrophoretic mobility shift assay demonstrated that yatein interrupted the formation of alpha-trans-induction factor/C1/Oct-1/GARAT multiprotein complex. The mechanisms of antiviral action of yatein seem to be mediated, by inhibiting HSV-1 alpha gene expression, including expression of the ICP0 and ICP4 genes, and by arresting HSV-1 DNA synthesis and structural protein expression in HeLa cells. These results suggest that yatein is an antiviral agent against HSV-1 replication. PMID:16540181

Kuo, Yuh-Chi; Kuo, Yueh-Hsiung; Lin, Yuang-Lian; Tsai, Wei-Jern

2006-02-20

339

Inhibition of signal transducer and activator of transcription 3 and cyclooxygenase-2 is involved in radiosensitization of cepharanthine in HeLa cells.  

UK PubMed Central (United Kingdom)

OBJECTIVES: To investigate the radiosensitizing effects of cepharanthine (CEP) in the human cervical adenocarcinoma HeLa cell line and to examine the underlying mechanisms. MATERIALS/METHODS: Survival of HeLa cells after treatment with or without ionizing radiation (IR) and CEP administration was investigated. MTT assays and apoptosis analysis were used to assess cytotoxicity. Nude mouse xenografts were established to evaluate the antitumor effects of CEP and IR in vivo. Expression of signal transducer and activator of transcription 3 (STAT3) and its downstream signaling molecules as well as cyclooxygenase-2 (COX-2) were examined by Western blot analysis. RESULTS: Clonogenic assays showed that treatment with CEP and IR resulted in significant radiosensitization. Cepharanthine and IR treatment achieved maximum cytotoxic effects on HeLa cells with regard to apoptosis induction. Cepharanthine efficiently decreased IR-induced STAT3 and COX-2 activation. The STAT3 target genes, including the antiapoptotic Bcl-2 and the cell cycle regulator c-Myc, were decreased concomitantly. In vivo administration of CEP (20 mg/kg every 2 days) combined with radiation in HeLa xenografts enhanced tumor growth delay and apoptosis (indicated by activated caspase-3 Western blot analysis), with reduced expression of STAT3, Bcl-2, c-Myc, and COX-2. CONCLUSIONS: Cepharanthine was shown to induce radiation sensitization in HeLa cells in vitro and in vivo. The inhibitory effects of CEP on STAT3 signaling pathway and COX-2 help us to better understand the radiosensitization of CEP.

Fang ZH; Li YJ; Chen Z; Wang JJ; Zhu LH

2013-05-01

340

Is high mobility group protein 17 phosphorylated in vivo? Re-examination of the HeLa cell cycle data.  

UK PubMed Central (United Kingdom)

When in vivo [32P] phosphate labeled HMG proteins from unsynchronized HeLa cells are separated by electrophoresis in acid-urea polyacrylamide gels, as opposed to separation in SDS-polyacrylamide, HMG 17 does not show any 32P incorporation. Likewise, no 32P radioactivity was found in HMG 17 protein isolated at different stages of the cell cycle from synchronized cells. By contrast, HMG 14 reveals a previously reported (Bhorjee, J.S. (1981) Proc. Natl. Acad. Sci. U.S.A. 78, 6944-6948) cell cycle stage-specific dependent phosphorylation with maximum 32P radioactivity in the G2 phase relative to G1. Furthermore, HMG 14 is resolved into multiple electrophoretic forms as phosphoprotein in the acid-urea system. The results presented seriously question the data on the in vivo phosphorylation of HMG 17, and suggest that these be reevaluated.

Bhorjee JS; Mellon I; Kifle L

1983-03-01

 
 
 
 
341

Identification of a set of miRNAs differentially expressed in transiently TIA-depleted HeLa cells by genome-wide profiling.  

UK PubMed Central (United Kingdom)

BACKGROUND: T-cell intracellular antigen (TIA) proteins function as regulators of cell homeostasis. These proteins control gene expression globally at multiple levels in response to dynamic regulatory changes and environmental stresses. Herein we identified a micro(mi)RNA signature associated to transiently TIA-depleted HeLa cells and analyzed the potential role of miRNAs combining genome-wide analysis data on mRNA and miRNA profiles. RESULTS: Using high-throughput miRNA expression profiling, transient depletion of TIA-proteins in HeLa cells was observed to promote significant and reproducible changes affecting to a pool of up-regulated miRNAs involving miR-30b-3p, miR125a-3p, miR-193a-5p, miR-197-3p, miR-203a, miR-210, miR-371-5p, miR-373-5p, miR-483-5p, miR-492, miR-498, miR-503-5p, miR-572, miR-586, miR-612, miR-615-3p, miR-623, miR-625-5p, miR-629-5p, miR-638, miR-658, miR-663a, miR-671-5p, miR-769-3p and miR-744-5p. Some up-regulated and unchanged miRNAs were validated and previous results confirmed by reverse transcription and real time PCR. By target prediction of the miRNAs and combined analysis of the genome-wide expression profiles identified in TIA-depleted HeLa cells, we detected connections between up-regulated miRNAs and potential target genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) database analysis suggest that target genes are related with biological processes associated to the regulation of DNA-dependent transcription, signal transduction and multicellular organismal development as well as with the enrichment of pathways involved in cancer, focal adhesion, regulation of actin cytoskeleton, endocytosis and MAPK and Wnt signaling pathways, respectively. When the collection of experimentally defined differentially expressed genes in TIA-depleted HeLa cells was intersected with potential target genes only 7 out of 68 (10%) up- and 71 out of 328 (22%) down-regulated genes were shared. GO and KEGG database analyses showed that the enrichment categories of biological processes and cellular pathways were related with innate immune response, signal transduction, response to interleukin-1, glomerular basement membrane development as well as neuroactive ligand-receptor interaction, endocytosis, lysosomes and apoptosis, respectively. CONCLUSION: All this considered, these observations suggest that individual miRNAs could act as potential mediators of the epigenetic switch linking transcriptomic dynamics and cell phenotypes mediated by TIA proteins.

Sánchez-Jiménez C; Carrascoso I; Barrero J; Izquierdo JM

2013-01-01

342

Apoptosis in HeLa cell exposed to different dose, dose-rate of 32P ?-irradiation and the correlation with cell-killing efficacy  

International Nuclear Information System (INIS)

[en] In an attempt to elucidate some aspects of the radiobiological basis of targeted radiotherapy in oncology, the apoptosis occurred have been studied in Hela cell lines after exposing to different doses and dose-rate radiation of 32P and the relationship between apoptosis occurred and the capacity of cell proliferation, which might be of help to the understanding of targeted radiotherapy. Asynchronous Hela cells were exposed to ? radiation from 32P absorbed in filter papers which were put closely under culture dishes of growing monolayer of Hela cell. The radiation response characteristics to different dose, dose-rate and radiation time were evaluated through cell-proliferation assessed by the colony-forming assay, cell cycle perturbation studied by flow cytometry and quantity analysis of apoptosis analyzed by flow cytometry and fluorescence microscopy. Morphological and flow cytometry analysis showed a delayed apoptosis. The programmed cell death approached a plateau between 48-72h post-irradiation. Electron and fluorescence microscopic studies showed the presence of morphologically apoptotic cells. Single dose radiation showed a higher apoptosis ratio than multiple low dose radiation, which did not correlate with clonal-forming assay, suggesting apoptosis ratio at a near time point post-irradiation is not a convincing indicator of radiation efficacy in the current experimental setting

2001-01-01

343

Differential effects of calcium antagonists on ABCG2/BCRP-mediated drug resistance and transport in SN-38-resistant HeLa cells.  

UK PubMed Central (United Kingdom)

The effects of 9 calcium antagonists on ABCG2/BCRP-mediated resistance and transport were examined in HeLa and SN-38-resistant HeLa (HeLa/SN100) cells, overexpressing ABCG2/BCRP. Sensitivity to mitoxantrone, an ABCG2/BCRP substrate, in HeLa/SN100 cells was significantly reversed by the coexistence of the calcium antagonists, except for diltiazem and verapamil. The accelerated transport activity of Hoechst33342, an ABCG2/BCRP substrate, in HeLa/SN100 cells was significantly decreased by the presence of the calcium antagonists, except for diltiazem, nifedipine or verapamil, returning to the level of HeLa cells. The present study classifies the calcium antagonists into 3 categories: strong (benidipine, felodipine, nicardipine, nisoldipine and nitrendipine), moderate (amlodipine and nifedipine) and weak (diltiazem and verapamil) inhibitors of ABCG2/BCRP.

Takara K; Matsubara M; Yamamoto K; Minegaki T; Takegami S; Takahashi M; Yokoyama T; Okumura K

2012-03-01

344

Differential effects of calcium antagonists on ABCG2/BCRP-mediated drug resistance and transport in SN-38-resistant HeLa cells.  

Science.gov (United States)

The effects of 9 calcium antagonists on ABCG2/BCRP-mediated resistance and transport were examined in HeLa and SN-38-resistant HeLa (HeLa/SN100) cells, overexpressing ABCG2/BCRP. Sensitivity to mitoxantrone, an ABCG2/BCRP substrate, in HeLa/SN100 cells was significantly reversed by the coexistence of the calcium antagonists, except for diltiazem and verapamil. The accelerated transport activity of Hoechst33342, an ABCG2/BCRP substrate, in HeLa/SN100 cells was significantly decreased by the presence of the calcium antagonists, except for diltiazem, nifedipine or verapamil, returning to the level of HeLa cells. The present study classifies the calcium antagonists into 3 categories: strong (benidipine, felodipine, nicardipine, nisoldipine and nitrendipine), moderate (amlodipine and nifedipine) and weak (diltiazem and verapamil) inhibitors of ABCG2/BCRP. PMID:22200670

Takara, Kohji; Matsubara, Mika; Yamamoto, Kazuhiro; Minegaki, Tetsuya; Takegami, Shigehiko; Takahashi, Minoru; Yokoyama, Teruyoshi; Okumura, Katsuhiko

2011-12-22

345

Xeroderma pigmentosum variant (XP-V) correcting protein from HeLa cells has a thymine dimer bypass DNA polymerase activity.  

Science.gov (United States)

Xeroderma pigmentosum variant (XP-V) represents one of the most common forms of this cancer-prone DNA repair syndrome. Unlike classical XP cells, XP-V cells are normal in nucleotide excision repair but defective in post-replication repair. The precise molecular defect in XP-V is currently unknown, but it appears to be a protein involved in translesion synthesis. Here we established a sensitive assay system using an SV40 origin-based plasmid to detect XP-V complementation activity. Using this system, we isolated a protein from HeLa cells capable of complementing the defects in XP-V cell extracts. The protein displays novel DNA polymerase activity which replicates cyclobutane pyrimidine dimer-containing DNA templates. The XPV polymerase activity was dependent on MgCl2, sensitive to NEM, moderately sensitive to KCl, resistant to both aphidicolin and ddTTP, and not stimulated by PCNA. In glycerol density gradients, the activity co-sedimented with a 54 kDa polypeptide at 3.5S, indicating that the monomeric form of this polypeptide was responsible for the activity. The protein factor corrected the translesion defects of extracts from three XPV cell strains. Bypass DNA synthesis by the XP-V polymerase occurred only in the presence of dATP, indicating that it can incorporate only dATP to bypass a di-thymine lesion. PMID:10369688

Masutani, C; Araki, M; Yamada, A; Kusumoto, R; Nogimori, T; Maekawa, T; Iwai, S; Hanaoka, F

1999-06-15

346

Preincubation of recombinant Ipa proteins of Shigella sonnei promotes entry of non-invasive Escherichia coli into HeLa cells.  

UK PubMed Central (United Kingdom)

Invasion plasmid antigens of Shigella sonnei, IpaB, C, D, were expressed as fusion proteins either with maltose-binding protein (MBP) or Strept-tag sequence. Affinity-purified IpaB and IpaD were separated from MBP by digestion with Factor Xa. Recombinant IpaC having Strept-tag sequence at its C-terminal was also purified by avidin affinity column chromatography. These recombinant proteins showed the ability to cause non-invasive Escherichia coli K-12 to internalize HeLa cell only when all of the proteins were preincubated with the bacterial prior to the inoculation of the mixture into HeLa cell culture. Electron microscopy also showed internalized bacteria within HeLa cells suggesting that functional complex of invasins (IpaB, C and D) were formed in vitro.

Terajima J; Moriishi E; Kurata T; Watanabe H

1999-10-01

347

Specific transcription of an adenoviral gene that possesses no TATA sequence homology in extracts of HeLa cells.  

UK PubMed Central (United Kingdom)

Transcription of the adenovirus type 2 (Ad2) IVa2 gene, which contains no TATA-like sequence in the region immediately upstream of the IVa2 cap sites (Baker, C. C., and Ziff, E. B. (1981) J. Mol. Biol. 149, 189-221), has been examined in extracts of HeLa cells (Manley, J. L., Fire, A., Cano, A., Sharp, P. A., and Gefter, M.L. (1980) Proc. Natl. Acad. Sci. U.S.A. 77, 3855-3859). Run-off transcripts of the predicted length of those initiated at the IVa2 cap sites were synthesized from different Ad2 DNA templates, each of which also contained the major late transcriptional control region. Mapping of the 5' ends of the RNA made from one template by a nuclease protection assay established the fidelity of initiation of IVa2 transcription in vitro. The efficiency of IVa2 expression in whole HeLa extracts was influenced quite dramatically by monovalent and divalent metal ion concentrations and the concentration of extract protein present in the reaction mixture. Under certain conditions, IVa2 run-off transcripts were made almost as efficiently as those from the Ad2 major late transcriptional control region. However, conditions promoting optimal IVa2 transcription in vitro did not favor recognition of the major late transcriptional control region, and vice versa: the synthesis of IVa2 and major late run-off transcripts responded differently to all parameters tested.

Leong K; Flint SJ

1984-09-01

348

3,4-dihydroxyphenyl acetic acid and (+)-epoxydon isolated from marine algae-derived microorganisms induce down regulation of epidermal growth factor activated mitogenic signaling cascade in Hela cells.  

UK PubMed Central (United Kingdom)

BACKGROUND: Epidermal growth factor receptor (EGFR) is a member of the receptor tyrosine kinase (RTK) family. Epidermal growth factor induces its dimerization and stimulates phosphorylation of intracellular tyrosine residues. Phosphorylation of EGFR is studied for cancer therapy because EGFR regulates many cellular processes including cell proliferation, differentiation, and survival. Hence, down-regulation of EGFR kinase activity results in inhibition of signaling cascades amenable for proliferation and progression of cell cycle. METHODS: In the study, we purified 3,4-dihydroxyphenyl acetic acid and (+)-epoxydon from Aspergillus sp. isolated from marine brown alga Ishige okamurae and Phoma herbarum isolated from marine red alga Hypnea saidana respectively and determined its anti-tumor activities against HeLa human cervical cancer cells. RESULTS: Two compounds suppressed EGFR activity in vitro with IC50 values for 3,4-dihydroxyphenyl acetic acid and (+)-epoxydon were 2.8 and 0.6 ?g/mL respectively and reduced the viable numbers of HeLa cells. Immunoblotting analysis exhibited that the compounds induced inhibition of cell growth by causing downregulation of the mitogenic signaling cascade, inactivation of p90RSK, and release of cytochrome c from mitochondria. CONCLUSIONS: Results suggest that decreased expression of active EGFR and EGFR-related downstream molecules by treatment with the compounds may results in the inhibition of cell growth and inducement of apoptosis.

Jo MJ; Bae SJ; Son BW; Kim CY; Kim GD

2013-01-01

349

Evidence from uv transcription mapping in HeLa cells that heterogeneous nuclear RNA is the messenger RNA precursor  

International Nuclear Information System (INIS)

[en] The effects of uv irradiation on the incorporation of [3H]uridine in HeLa (human) cell mRNA, rRNA, heterogeneous nuclear RNA (hnRNA) and early mRNA from adenovirus type 2 have been compared. The uv target size of cell mRNA is at least 3 times larger than the average size of the mRNA itself and larger than the adenovirus-2 early mRNA, which is known to derive from transcription units of about 1.5-5.0 kilobases. The uv target size of hnRNA, in contrast, is about the same as its size determined by sedimentation and overlaps with the target size of mRNA. It is concluded that most mRNA derives from a higher molecular weight hnRNA molecule

1977-01-01

350

Cleavage of Eukaryotic Translation Initiation Factor 4G by Exogenously Added Hybrid Proteins Containing Poliovirus 2Apro in HeLa Cells: Effects on Gene Expression  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Efficient cleavage of both forms of eukaryotic initiation factor 4G (eIF4G-1 and eIF4G-2) has been achieved in HeLa cells by incubation with hybrid proteins containing poliovirus 2Apro. Entry of these proteins into cells is promoted by adenovirus particles. Substantial levels of ongoing translation ...

Novoa, Isabel; Carrasco, Luis

351

Heat-enhanced reactivation of UV-irradiated adenovirus 2 is not associated with enhanced mutagenesis in HeLa cells  

Energy Technology Data Exchange (ETDEWEB)

The reversion frequency of an adenovirus 2 temperature-sensitive growth mutant irradiated with different doses of UV light was determined after infection of control, UV-irradiated and heat-shocked HeLa cells. No enhancement of mutagenesis by treatment of the cells was observed. Heat-enhanced viral reactivation does not therefore display a significant error-prone component.

Piperakis, S.M.; McLennan, A.G. (Liverpool Univ. (UK). Dept. of Biochemistry)

1984-04-01

352

Heat-enhanced reactivation of UV-irradiated adenovirus 2 is not associated with enhanced mutagenesis in HeLa cells  

International Nuclear Information System (INIS)

The reversion frequency of an adenovirus 2 temperature-sensitive growth mutant irradiated with different doses of UV light was determined after infection of control, UV-irradiated and heat-shocked HeLa cells. No enhancement of mutagenesis by treatment of the cells was observed. Heat-enhanced viral reactivation does not therefore display a significant error-prone component. (orig.).

1984-