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1

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.

Liu Xianting

2010-01-01

2

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

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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. PMID:23925245

Adey, Andrew; Burton, Joshua N; Kitzman, Jacob O; Hiatt, Joseph B; Lewis, Alexandra P; Martin, Beth K; Qiu, Ruolan; Lee, Choli; Shendure, Jay

2013-08-01

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HIF-1 and NDRG2 contribute to hypoxia-induced radioresistance of cervical cancer Hela cells  

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Hypoxia inducible factor 1 (HIF-1), the key mediator of hypoxia signaling pathways, has been shown involved in hypoxia-induced radioresistance. However, the underlying mechanisms are unclear. The present study demonstrated that both hypoxia and hypoxia mimetic cobalt chloride could increase the radioresistance of human cervical cancer Hela cells. Meanwhile, ectopic expression of HIF-1 could enhance the resistance of Hela cells to radiation, whereas knocking-down of HIF-1 could increase the sensitivity of Hela cells to radiation in the presence of hypoxia. N-Myc downstream-regulated gene 2 (NDRG2), a new HIF-1 target gene identified in our lab, was found to be upregulated by hypoxia and radiation in a HIF-1-dependent manner. Overexpression of NDRG2 resulted in decreased sensitivity of Hela cells to radiation while silencing NDRG2 led to radiosensitization. Moreover, NDRG2 was proved to protect Hela cells from radiation-induced apoptosis and abolish radiation-induced upregulation of Bax. Taken together, these data suggest that both HIF-1 and NDRG2 contribute to hypoxia-induced tumor radioresistance and that NDRG2 acts downstream of HIF-1 to promote radioresistance through suppressing radiation-induced Bax expression. It would be meaningful to further explore the clinical application potential of HIF-1 and NDRG2 blockade as radiosensitizer for tumor therapy.

2010-07-15

4

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

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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. PMID:24064712

Han, Bo Ram; You, Bo Ra; Park, Woo Hyun

2013-12-01

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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

2008-01-01

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

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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. PMID:22989702

Momtaz, S; Hussein, A A; Ostad, S N; Abdollahi, M; Lall, N

2013-01-01

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Effect of cerium lanthanide on Hela and MCF-7 cancer cell growth in the presence of transferring  

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The anti-cancer activity of metal ions in the lanthanide group is being considered recently. It has been reported that cerium salts might stimulate the metabolism and therefore, produce anti-cancer effects. However, little is known about the effects of protein-cerium complex in controlling cancer cell growth. The aim of the present study was to elucidate the possible pathways for the cytotoxic effect of cerium in the presence of apo-transferrin on two cancer cell lines (Hela and MCF-7), that ...

Palizban, A. A.; Sadeghi-aliabadi, H.; Abdollahpour, F.

2010-01-01

8

Study on the effects of anti-proliferative protein Tob1 on the radio-sensitivity of human cervix cancer cell line HeLa  

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In order to investigate the effects of human anti-proliferative protein Tob1 (transducer of ErbB2, 1) on the radio-sensitivity of human cervix cancer cells HeLa, the full length Tob1 recombinant plasmid pcDNA3.0/Tob1 (pc3/Tob1) was constructed and transfected into HeLa cells by lipofectamine. And G418 selection was used to get HeLa cell line stably expressed exogenous Tob1. The clonogenic assay was applied to study the effects of Tob1 on HeLa cell radio-sensitivity, while flow cytometry assay and Western Blot analysis were performed to determine the related mechanisms. It was shown that,compared with parental HeLa cells and mock plasmid pcDNA3.0 transfected HeLa/pc3, the radio-sensitivity of HeLa cells transfected with Tob1 was increased obviously. It was also found that increased Tob1 expression could enhance cell apoptosis of HeLa induced by irradiation, while up-regulated protein expression level of Bax,but down-regulated Bcl2 expression occur at the same time. These results suggested that Tob1 might play a role as radio-sensitizer of cervix cancer cell line HeLa via regulating the expression of apoptosis related genes. (authors)

2010-08-01

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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

2012-10-01

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Study on the effect of artesunate combined with irradiation on DNA damage of HeLa and Siha cells of human cervical cancer  

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In order to investigate the effect of artesunate combined with irradiation on DNA damage of HeLa and Siha cells of human cervical cancer, HeLa and Siha cells were cultured in vitro and exposed to different concentration of artesunate for 24 h and MTT assay was used to observe the inhibitory effect of different concentration of artesunate on the proliferation of HeLa and Siha cells. The cells were divided into 2 groups as the irradiated group and the union treatment group. Here it was set up four absorbed doses of 60Co ?-irradiation in each group with 0, 2, 4 and 6 Gy, and the DNA damage were detected by single cell gel electrophoresis assay. MTT analysis showed that the inhibition of artesunate on HeLa and Siha cells of cervical cancer was in concentration-dependent manners. Single cell gel electrophoresis showed that the DNA damage of HeLa cells treated with artesunate was more serious than that treated only with irradiation (P<0.05), but had no such effect on Siha cells. Artesunate can increase the radio-sensitivity of HeLa cells cervical cancer with p53 mutant, but has no such effect on wide type p53 cells. (authors)

2011-02-01

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

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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< 0.05 or P< 0.01). The inhibiting effect of 6 Gy combined with autophagy inducer rapamycin on the proliferation of Hela cells weakened (P< 0.05). The inhibiting effects of 6 Gy combined with autophagy inhibitor 3-MA on the cell proliferation were higher than those in 6 Gy group (P< 0.05). Conclusion: Ionizing radiation combined with autophagy inducers can inhibit apoptosis in Hela cells, while the ionizing radiation combined with autophagy inhibitors can promote their apoptosis. (authors)

2010-09-01

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Radiosensitization effect of folate-conjugated gold nanoparticles on HeLa cancer cells under orthovoltage superficial radiotherapy techniques  

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Due to the high atomic number of gold nanoparticles (GNPs), they are known as new radiosensitizer agents for enhancing the efficiency of superficial radiotherapy techniques by increasing the dose absorbed in tumor cells wherein they can be accumulated selectively. The aim of this study was to compare the effect of various common low energy levels of orthovoltage x-rays and megavoltage ?-rays (Co-60) on enhancing the therapeutic efficiency of HeLa cancer cells in the presence of conjugated folate and non-conjugated (pegylated) GNPs. To achieve this, GNPs with an average diameter of 52 nm were synthesized and conjugated to folic acid molecules. Pegylated GNPs with an average diameter of 47 nm were also synthesized and used as non-conjugated folate GNPs. Cytotoxicity assay of the synthesized folate-conjugated and pegylated GNPs was performed using different levels of nanoparticle concentration incubated with HeLa cells for 24 h. The radiosensitizing effect of both the conjugated and pegylated GNPs on the cells at a concentration of 50 µM was compared using MTT as well as clonogenic assays after exposing them to 2 Gy ionizing radiation produced by an orthovoltage x-ray machine at four different kVps and ?-rays of a Co-60 unit. Significant differences were noted among various irradiated groups with and without the folate conjugation, with an average dose enhancement factor (DEF) of 1.64 ± 0.05 and 1.35 ± 0.05 for the folate-conjugated and pegylated GNPs, respectively. The maximum DEF was obtained with the 180 kVp x-ray beam for both of the GNPs. Folate-conjugated GNPs can significantly enhance the cell killing potential of orthovoltage x-ray energies (especially at 180 kVp) in folate receptor-expressing cancer cells, such as HeLa, in superficial radiotherapy techniques.

Khoshgard, Karim; Hashemi, Bijan; Arbabi, Azim; Javad Rasaee, Mohammad; Soleimani, Masoud

2014-05-01

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Radiosensitization effect of folate-conjugated gold nanoparticles on HeLa cancer cells under orthovoltage superficial radiotherapy techniques.  

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Due to the high atomic number of gold nanoparticles (GNPs), they are known as new radiosensitizer agents for enhancing the efficiency of superficial radiotherapy techniques by increasing the dose absorbed in tumor cells wherein they can be accumulated selectively. The aim of this study was to compare the effect of various common low energy levels of orthovoltage x-rays and megavoltage ?-rays (Co-60) on enhancing the therapeutic efficiency of HeLa cancer cells in the presence of conjugated folate and non-conjugated (pegylated) GNPs. To achieve this, GNPs with an average diameter of 52 nm were synthesized and conjugated to folic acid molecules. Pegylated GNPs with an average diameter of 47 nm were also synthesized and used as non-conjugated folate GNPs. Cytotoxicity assay of the synthesized folate-conjugated and pegylated GNPs was performed using different levels of nanoparticle concentration incubated with HeLa cells for 24 h. The radiosensitizing effect of both the conjugated and pegylated GNPs on the cells at a concentration of 50 µM was compared using MTT as well as clonogenic assays after exposing them to 2 Gy ionizing radiation produced by an orthovoltage x-ray machine at four different kVps and ?-rays of a Co-60 unit. Significant differences were noted among various irradiated groups with and without the folate conjugation, with an average dose enhancement factor (DEF) of 1.64 ± 0.05 and 1.35 ± 0.05 for the folate-conjugated and pegylated GNPs, respectively. The maximum DEF was obtained with the 180 kVp x-ray beam for both of the GNPs. Folate-conjugated GNPs can significantly enhance the cell killing potential of orthovoltage x-ray energies (especially at 180 kVp) in folate receptor-expressing cancer cells, such as HeLa, in superficial radiotherapy techniques. PMID:24733041

Khoshgard, Karim; Hashemi, Bijan; Arbabi, Azim; Rasaee, Mohammad Javad; Soleimani, Masoud

2014-05-01

14

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

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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. PMID:23042479

Xiao, Wei; Zhou, Shumin; Xu, Hua; Li, Heng; He, Guoqing; Liu, Yingle; Qi, Yipeng

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.  

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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

2011-04-01

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The Ability to Survive Mitosis in the Presence of Microtubule Poisons Differs Significantly Between Human Nontransformed (RPE-1) and Cancer (U2OS, HeLa) Cells  

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We used live cell imaging to compare the fate of human nontransformed (RPE-1) and cancer (HeLa, U2OS) cells as they entered mitosis in nocodazole or taxol. In the same field, and in either drug, a cell in all lines could die in mitosis, exit mitosis and die within 10 h, or exit mitosis and survive ?10 h. Relative to RPE-1 cells, significantly fewer HeLa or U2OS cells survived mitosis or remained viable after mitosis: in nocodazole concentrations that inhibit spindle microtubule assembly, or...

Brito, Daniela A.; Rieder, Conly L.

2009-01-01

17

Different effects of adenylyl cyclase activators and phosphodiesterases inhibitors on cervical cancer (HeLa) and breast cancer (MCF-7) cells proliferation.  

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Breast and cervical cancers are the most common cancers in Iran and worldwide. Hormonal stimulation of cyclic adenosine mono phosphate (cAMP) and the cAMP-dependent protein kinase PKA regulates cell growth by different mechanism. cAMP can stimulate cell growth in many cell types while inhibiting cell growth in others. In some cell lines have been shown that the proliferation of tumor cells is reduced by increasing cAMP in cells. In this study, we evaluate growth arrest of selective PDE3 and non-selective PDE inhibitors, which lead to increase level of cAMP in cervical (HeLa) and breast cancer (MCF7) cell lines have been studied. Cells were incubated with different concentrations of selective, non-selective PDE inhibitors, beta adrenergic receptor agonist and direct stimulator of adenylyl cyclase. Cell viability was quantitated by MTT assay. Apoptotic cells were determined using PI staining of DNA fragmentation by flow cytometry (sub-G1 peak). Result showed that selective PDE inhibitors decreased cell viability in HeLa and MCF-7 cells as a time-dependent manner. Non-selective inhibitor and beta-adrenergic receptor agonist also decrease cell viability but they are less powerful than selective PDE3 inhibitors. Forskolin had no effect in viability of cells. Analysis of DNA fragmentation by flow cytometry showed apoptosis involved in selective PDE3 inhibitors induced toxicity in HeLa cell. Thus, the growth inhibitory effects of selective PDE3 inhibitors are more effective than non-selective inhibitor. Further studies are needed to investigate the mechanism of action is on the field. PMID:24593874

Mahdian, Davood; Shafiee-Nick, Reza; Mousavi, Seyed Hadi

2014-05-01

18

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

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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...

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

2010-01-01

19

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

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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

20

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

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Full Text Available Abstract 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 Charlotte M

2013-01-01

 
 
 
 
21

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

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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

2011-04-01

22

Requirement of T-lymphokine-activated killer cell-originated protein kinase for TRAIL resistance of human HeLa cervical cancer cells  

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T-lymphokine-activated killer cell-originated protein kinase (TOPK) appears to be highly expressed in various cancer cells and to play an important role in maintaining proliferation of cancer cells. However, the underlying mechanism by which TOPK regulates growth of cancer cells remains elusive. Here we report that upregulated endogenous TOPK augments resistance of cancer cells to apoptosis induced by tumor necrosis factor-related apoptosis inducing ligand (TRAIL). Stable knocking down of TOPK markedly increased TRAIL-mediated apoptosis of human HeLa cervical cancer cells, as compared with control cells. Caspase 8 or caspase 3 activities in response to TRAIL were greatly incremented in TOPK-depleted cells. Ablation of TOPK negatively regulated TRAIL-mediated NF-{kappa}B activity. Furthermore, expression of NF-{kappa}B-dependent genes, FLICE-inhibitory protein (FLIP), inhibitor of apoptosis protein 1 (c-IAP1), or X-linked inhibitor of apoptosis protein (XIAP) was reduced in TOPK-depleted cells. Collectively, these findings demonstrated that TOPK contributed to TRAIL resistance of cancer cells via NF-{kappa}B activity, suggesting that TOPK might be a potential molecular target for successful cancer therapy using TRAIL.

Kwon, Hyeok-Ran [Department of Biochemistry, College of Medicine, Konyang University, 685 Gasuwon-dong, Seo-gu, Daejeon 302-718 (Korea, Republic of); Lee, Ki Won [Department of Bioscience and Biotechnology, Konkuk University, 1 Hwayang-dong, Gwangjin-gu, Seoul 143-701 (Korea, Republic of); Dong, Zigang [Hormel Institute, University of Minnesota, 801 16th Avenue NE, Austin, MN 55912 (United States); Lee, Kyung Bok [Department of Biochemistry, College of Medicine, Konyang University, 685 Gasuwon-dong, Seo-gu, Daejeon 302-718 (Korea, Republic of); Oh, Sang-Muk, E-mail: sangmuk_oh@konyang.ac.kr [Department of Biochemistry, College of Medicine, Konyang University, 685 Gasuwon-dong, Seo-gu, Daejeon 302-718 (Korea, Republic of)

2010-01-01

23

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.  

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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. PMID:24008363

Kim, Hye-Sun; Yi, Bo-Rim; Hwang, Kyung-A; Kim, Seung U; Choi, Kyung-Chul

2013-10-01

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The neem limonoids azadirachtin and nimbolide induce cell cycle arrest and mitochondria-mediated apoptosis in human cervical cancer (HeLa) cells.  

Science.gov (United States)

Limonoids from the neem tree (Azadirachta indica) have attracted considerable research attention in recent years owing to their potent antioxidant and anti-proliferative effects. The present study was designed to investigate the cellular and molecular mechanisms by which azadirachtin and nimbolide exert cytotoxic effects in the human cervical cancer (HeLa) cell line. Both azadirachtin and nimbolide significantly suppressed the viability of HeLa cells in a dose-dependent manner by inducing cell cycle arrest at G0/G1 phase accompanied by p53-dependent p21 accumulation and down-regulation of the cell cycle regulatory proteins cyclin B, cyclin D1 and PCNA. Characteristic changes in nuclear morphology, presence of a subdiploid peak and annexin-V staining pointed to apoptosis as the mode of cell death. Increased generation of reactive oxygen species with decline in the mitochondrial transmembrane potential and release of cytochrome c confirmed that the neem limonoids transduced the apoptotic signal via the mitochondrial pathway. Altered expression of the Bcl-2 family of proteins, inhibition of NF-kappaB activation and over-expression of caspases and survivin provide compelling evidence that azadirachtin and nimbolide induce a shift of balance toward a pro-apoptotic phenotype. Antioxidants such as azadirachtin and nimbolide that can simultaneously arrest the cell cycle and target multiple molecules involved in mitochondrial apoptosis offer immense potential as anti-cancer therapeutic drugs. PMID:20429769

Priyadarsini, R Vidya; Murugan, R Senthil; Sripriya, P; Karunagaran, D; Nagini, S

2010-06-01

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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  

Directory of Open Access Journals (Sweden)

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.

K Pooshang Bagheri

2013-06-01

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Irradiation And Papillomavirus E2 Proteins On Hela Cells  

International Nuclear Information System (INIS)

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

2005-04-01

27

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  

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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 li...

Ehsan Oskoueian; Norhani Abdullah; Syahida Ahmad

2012-01-01

28

Juglans mandshurica Maxim extracts exhibit antitumor activity on HeLa cells in vitro.  

Science.gov (United States)

The present study examined the potential application of Juglans mandshurica Maxim extracts (HT) for cancer therapy by assessing their anti?proliferative activity, reduction of telomerase activity, induction of apoptosis and cell cycle arrest in S phase in HeLa cells. From the perspective of using HT as a herbal medicine, photomicroscopy and florescent microscopy techniques were utilized to characterize the effect of the extracts on telomerase activity and cell morphology. Flow cytometry was employed to study apoptosis and cell cycle of HeLa cells, and DNA laddering was performed. The results showed that HT inhibited cell proliferation and telomerase activity, induced apoptosis and caused S phase arrest of HeLa cells in vitro. HT inhibited HeLa cell proliferation significantly, and the highest inhibition rate was 83.7%. A trap?silver staining assay showed that HT was capable of markedly decreasing telomerase activity of HeLa cells and this inhibition was enhanced in a time? and dose?dependent manner. Results of a Hoechst 33258 staining assay showed that HeLa cells treated by HT induced cell death. Through DNA agarose gel electrophoresis, DNA ladders of HeLa cells treated with HT were observed, indicating apoptosis. In conclusion, the present study demonstrated that HT exhibited anti?tumor effects comprising the inhibition of growth and telomerase activity as well as apoptosis and cell cycle arrest in HeLa cells. PMID:24566804

Xin, Nian; Hasan, Murtaza; Li, Wei; Li, Yan

2014-04-01

29

Biofabrication of Ag nanoparticles using Sterculia foetida L. seed extract and their toxic potential against mosquito vectors and HeLa cancer cells.  

Science.gov (United States)

A one-step and eco-friendly process for the synthesis of silver-(protein-lipid) nanoparticles (Ag-PL NPs) (core-shell) has been developed using the seed extract from wild Indian Almond tree, Sterculia foetida (L.) (Sterculiaceae). The reaction temperature played a major role in controlling the size and shell formation of NPs. The amount of NPs synthesized and qualitative characterization was done by UV-vis spectroscopy and transmission electron microscopy (TEM), respectively. TEM studies exhibited controlled dispersity of spherical shaped NPs with an average size of 6.9±0.2nm. Selected area electron diffraction (SAED) and X-ray diffraction (XRD) revealed 'fcc' phase and crystallinity of the particles. X-ray photoelectron spectroscopy (XPS) was used to identify the protein-lipid (PL) bilayer that appears as a shell around the Ag core particles. The thermal stability of the Ag-PL NPs was examined using thermogravimetric analysis (TGA). Further analysis was carried out by using Fourier transform infrared spectroscopy (FTIR), where the spectra provided evidence for the presence of proteins and lipid moieties ((2n-octylcycloprop-1-enyl)-octanoic acid (I)), and their role in synthesis and stabilization of Ag NPs. This is the first report of plant seed assisted synthesis of PL conjugated Ag NPs. These formed Ag-PL NPs showed potential mosquito larvicidal activity against Aedes aegypti (L.), Anopheles stephensi Liston and Culex quinquefasciatus Say. These Ag-PL NPs can also act as promising agents in cancer therapy. They exhibited anti-proliferative activity against HeLa cancer cell lines and a promising toxicity was observed in a dose dependent manner. Toxicity studies were further supported by the cellular DNA fragmentation in the Ag-PL NPs treated HeLa cells. PMID:24863217

Rajasekharreddy, Pala; Rani, Pathipati Usha

2014-06-01

30

Photodynamic Effects of Pterin on HeLa Cells  

DEFF Research Database (Denmark)

Pterins, heterocyclic compounds widespread in biological systems, participate in relevant biological processes and are able to act as photosensitizers. In the present study, we ascertained that 2-aminopteridin-4(3H)-one, abbreviated as Ptr, is readily incorporated into and � or onto cervical cancer cells (HeLa) and that these cells die upon UV-A irradiation of Ptr. Cell death was assessed using two tests: (1) the Rhodamine 123 fluorescence assay for mitochondrial viability and (2) the Trypan Blue assay for membrane integrity. The data suggest that, for Ptr-dependent photoinitiated cell death, events related to mitochondrial failure precede those associated with the failure of the cell membrane.

Denofrio, M. Paula; Lorente, Carolina

2011-01-01

31

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

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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...

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

2013-01-01

32

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

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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 demonst...

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

2008-01-01

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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.

Syahida Ahmad

2012-09-01

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english 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 o [...] f 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, José María Eloy; Vázquez-Chagoyán, Juan Carlos; Martínez-Castañeda, José Simón; Estrada-Franco, José Guillermo; Aparicio-Burgos, José Esteban; Acosta-Dibarrat, Jorge; Barbabosa-Pliego, Alberto.

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

Science.gov (United States)

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.

2013-01-01

36

Antiproliferative effects of some medicinal plants on HeLa cells  

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Full Text Available Medicinal plants maintain the health and vitality of individuals, and also have potential curative effect on various diseases, including cancer. In this study were investigated the antiproliferative effects of water extracts of previously obtained ethanolic dry extracts of three different medicinal plants (Echinacea angustifolia, Salvia officinalis and Melissa officinalis on cell lines derived from human cervix adenocarcinoma (HeLa cells. The best cytotoxic activity (IC50 = 43.52 ?g/ml on HeLa cell lines was exhibited by Echinacea angustifolia. The extract of Salvia officinalis also showed a good cytotoxic activity against HeLa cell lines; the IC50 value was 70.41 ?g/ml. Melissa officinalis manifested a slightly weaker cytotoxic activity and an IC50 value of 122.22 ?g/ml. [Projekat Ministarstva nauke Republike Srbije, br. 34021 i br. 175011

Ceni?-Miloševi? Desanka

2013-01-01

37

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

38

Growth regulation of HeLa cells by 1060 nm photons  

International Nuclear Information System (INIS)

Living organisms are open systems dominated by electromagnetic interaction. An essential feature of a living system is its cybernetic process which imply their capability of adaptation and sensitivity to internal and external fluctuations. The experimental results show that coherent and incoherent light of 1060 nm wavelength influences the metabolic processes and consequently the proliferation of cancer cell cultures (HeLa). Light induced regulation of HeLa cell growth depends on the cell density, the state of the cell culture and the amount of light irradiation. Best proliferation inhibiting effects can be obtained by application of 200 J/m2 on HeLa cells in Lag-Phase and a typical cell density of 5.104 cells/cm2. Proceeding on the singlet oxygen hypothesis (KLIMA, H. et al.; 1990), it is shown mathematically that the dynamical behaviour of the NADH model is influenced by 1060 nm photons. Both, the experimental and the numerical results support our hypothesis: 1060 nm photons regulate the proliferation of HeLa cells. (author)

1993-01-01

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Antiproliferative effects of some medicinal plants on HeLa cells  

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Medicinal plants maintain the health and vitality of individuals, and also have potential curative effect on various diseases, including cancer. In this study were investigated the antiproliferative effects of water extracts of previously obtained ethanolic dry extracts of three different medicinal plants (Echinacea angustifolia, Salvia officinalis and Melissa officinalis) on cell lines derived from human cervix adenocarcinoma (HeLa cells). The best cytotoxic activity (IC50 = 43.52 ?g/...

Ceni?-Miloševi? Desanka; Tambur Z.; Bokonji? D.; Ivan?aji? S.; Stanojkovi? Tatjana; Grozdani? Nadja; Jurani? Zorica

2013-01-01

40

Energy metabolism in hela and walker-256 cells  

International Nuclear Information System (INIS)

Attempts to measure ATP content in terms of amino acid incorporation into proteins in the presence of oxamate with and without glucose seem to indicate that uptake of labelled amino acid is independent of ATP level. This was quite in contrast to actual growth measurements where growth inhibition was observed only in the presence of glucose. Probably oxamate interfered with the transport of amino acid into HeLa cells and this was readily releived by adding pyruvate. In another system using Walker 256 carcinoma cells, oxamate effect to interfere with the uptake of label either through incorporation or transport was not observed suggesting a difference between HeLa and Walker cells in energy potential. (author)

1973-02-01

 
 
 
 
41

Purification and reconstitution of HeLa cell microtubules.  

Science.gov (United States)

Microtubules from suspension cultures of HeLa cells have been purified by carrying them through four complete cycles of polymerization at 37 degrees C and depolymerization at 4 degrees C. These microtubules show, in addition to the major alpha- and beta-tubulin components, major proteins with molecular weights of 201 000-206 000 (comprising 4.5% of the total protein), proteins with molecular weights of 97 000, 100 000, 104 000, and 114 000 (together comprising approximately 2% of the total protein), and minor components with molecular weights of 68 000 and 151 000. HeLa microtubules have also been reconstituted from purified HeLa tubulin and proteins from HeLa microtubules separated from tubulin by DEAE-cellulose column chromatography. Experiments on the fractionation and reconstitution of both two- and four-cycle microtubules suggest that the 201 000-206 000-dalton proteins are incorporated into microtubules and promote tubulin polymerization. Microtubules formed by fractionationand reconstitution of two-cycle microtubules also contain several other proteins with molecular weights of 132 000, 146 000, 151 000, 160 000, and 284 000, although these are not present in microtubules carried through four assembly-disassembly cycles. Evidence is also presented which shows that a 68 000-dalton protein which is a prominent component of HeLa microtubules after two polymerization-depolymerization cycles does not stoichiometrically copurify with tubulin through repeated assembly--disassembly cycles and does not stimulate tubulin polymerization. On the other hand, the sedimentation of this 68 000-dalton protein is apparently influenced by the presence of polymerized microtubules, suggesting that this protein may be a component of a system whjich interacts weakly with microtubules. Finally, evidence is presented suggesting that two-cycle microtubules contain a proteolytic activity that can digest the 201 000-206 000-dalton proteins. PMID:7407082

Weatherbee, J A; Luftig, R B; Weihing, R R

1980-08-19

42

Purification of histone ubiquitin ligases from HeLa cells.  

Science.gov (United States)

Posttranslational histone modifications play an important role in regulating chromatin based nuclear processes including transcription. Of these modifications, histone ubiquitination is among the least understood. Histone ubiquitination predominately targets histones H2A and H2B. While ubiquitination of H2B is evolutionarily conserved from budding yeast to mammals, ubiquitination of H2A has not been detected in budding yeast, worms, or plants. Until recently, studies of histone ubiquitination lagged far behind the study of other histone modifications, largely because antibodies specific for ubiquitinated histones are difficult to generate. Despite this obstacle, the identification of the enzymatic machineries involved in histone ubiquitination, together with the successful use of a combination of genetic and immunoblot approaches to detect ubiquitinated histones, have helped to reveal important regulatory roles for this modification in transcriptional initiation and elongation, cell cycle progression, and DNA damage response. With the aid of the recently developed ubiquitinated histone-specific antibodies, an intriguing link between histone ubiquitination and cancer development has been established. While the enzymes involved in H2B ubiquitination were identified first in budding yeast and subsequently in higher organisms based on gene homology, the identification of the enzymatic machineries involved in H2A ubiquitination largely depended on a biochemical purification approach. The unbiased search for ubiquitin ligases targeting histones also led to the identification of a H3 and H4 ubiquitin ligase. Here we detail a protocol for the biochemical approach to identify histone ubiquitin ligase(s) from HeLa cells. Similar approaches have been successfully used to identify histone methyltransferases, histone demethylases, chromatin remodeling factors, and general transcription factors. So long as an in vitro enzymatic assay can be established, the approach we describe can be easily adapted to identify other histone and non-histone modifying enzymes. PMID:21402158

Jones, Amanda; Joo, Heui-Yun; Robbins, Woody; Wang, Hengbin

2011-07-01

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Biocompatibility of Porous Spherical Calcium Carbonate Microparticles on Hela Cells  

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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 ...

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

2012-01-01

44

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english 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, th [...] e 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.

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

45

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

46

LIV-1 suppression inhibits HeLa cell invasion by targeting ERK1/2-Snail/Slug pathway  

International Nuclear Information System (INIS)

It was reported that expression of the estrogen-regulated zinc transporter LIV-1 was particularly high in human cervical cancer cell line HeLa. This result prompted us to study the role that LIV-1 played in human cervical cancer. The results of real-time PCR showed that LIV-1 mRNA was significantly higher in cervical cancer in situ than in normal tissues. RNAi mediated suppression of LIV-1 in HeLa cells significantly inhibited cell proliferation, colony formation, migration, and invasive ability, but had no effect on cell apoptosis. Furthermore, LIV-1 suppression is accompanied by down-regulation of p44/42 MAPK, phospho-p44/42 MAPK, Snail and Slug expression levels. Hence, our data provide the first evidence that LIV-1 mRNA is overexpressed in cervical cancer in situ and is involved in invasion of cervical cancer cells through targeting MAPK-mediated Snail and Slug expression

2007-11-09

47

In vitro studies on radiosensitization effect of glucose capped gold nanoparticles in photon and ion irradiation of HeLa cells  

Energy Technology Data Exchange (ETDEWEB)

Highlights: ? Glucose capped gold nanoparticles (Glu-AuNPs) are synthesized for internalization in HeLa cells (cervical cancer cells). ? Internalization of Glu-AuNPs in HeLa cells is confirmed by cross section TEM of cells. ? Irradiation (by C ion or ?-rays) of HeLa cells with internalized Glu-AuNPs results in enhanced radiosensitization. ? There is about 30% reduction in radiation dose for 90% cell killing of HeLa cells, when internalized by Glu-AuNPs. ? The enhanced radiosensitization due to Glu-AuNPs is of interest for researchers in nanobiotechnology and radiation biology. -- Abstract: Noble metal nanoparticles are of great interest due to their potential applications in diagnostics and therapeutics. In the present work, we synthesized glucose capped gold nanoparticle (Glu-AuNP) for internalization in the HeLa cell line (human cervix cancer cells). The capping of glucose on Au nanoparticle was confirmed by Raman spectroscopy. The Glu-AuNP did not show any toxicity to the HeLa cell. The ?-radiation and carbon ion irradiation of HeLa cell with and without Glu-AuNP were performed to evaluate radiosensitization effects. The study revealed a significant reduction in radiation dose for killing the HeLa cells with internalized Glu-AuNPs as compared to the HeLa cells without Glu-AuNP. The Glu-AuNP treatment resulted in enhancement of radiation effect as evident from increase in relative biological effectiveness (RBE) values for carbon ion irradiated HeLa cells.

Kaur, Harminder; Pujari, Geetanjali [Radiation Biology Group, Inter University Accelerator Centre, Post Box 10502, New Delhi 110067 (India); Semwal, Manoj K. [Army Hospital (R and R), Delhi Cantonment, New Delhi 110010 (India); Sarma, Asitikantha [Radiation Biology Group, Inter University Accelerator Centre, Post Box 10502, New Delhi 110067 (India); Avasthi, Devesh Kumar, E-mail: dka@iuac.res.in [Radiation Biology Group, Inter University Accelerator Centre, Post Box 10502, New Delhi 110067 (India)

2013-04-15

48

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)

2006-10-01

49

The effect of uranyl acetate on human lymphoblastoid cells (RPMI 6410) and HeLa cells.  

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RPMI 6410 cells and HeLa cells were exposed to uranyl acetate. In RPMI 6410 cell cultures this produced single-membrane-bound presumably lysosomal bodies (called "uraniosomes") containing electron-dense crystals in the cultured cells and crystalline deposits in extracellular locations. Neither uraniosomes nor extracellular uranium deposits were found in HeLa cell cultures. All uraniosomes and extracellular uranium deposits analysed by electron-probed X-ray analysis were found to conta...

1982-01-01

50

Variation and adaptation of Pseudomonas aeruginosa toxicity to HeLa cells and fibroblasts.  

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The toxic components of supernatants from Pseudomonas aeruginosa cultures directed against HeLa cells and Staphylococcus aureus were evaluated with the aim of discovering interactions. Supernatants of eight different strains of P. aeruginosa were assayed for cytotoxic activity. All were active against HeLa cells; seven were toxic for S. aureus. On repeated suspension of P. aeruginosa in 0.9% sodium chloride solution, a shift from HeLa cell toxicity to staphylococcal lytic activity occurred al...

Mu?ller, H.; Kettelhack, C.; Kettelhack, M.; Sonntag, H. G.; Keilich, G.; Brossmer, R.; Richards, J.; Kinzel, V.; Ba?uerlein, E.; Pech, H.

1986-01-01

51

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

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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

52

From HeLa cell division to infectious diarrhoea  

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Hela S3 cells were grown in suspension both randomly and, synchronously using hydroxyurea which blocks cells at the G1/S interface. Cryosections were prepared, freeze-dried and analyzed by X-ray microanalysis. As cells moved into S and through M phases (Na) and (Cl) increased; both returned to normal levels upon re-entering G1 phase. The Na/K ratio was 1:1 in G1 phase. Infection of HeLa S3 cells in G1 phase with vaccinia virus resulted in no change in intracellular (Na). Infection of neonatal mice with murine rotavirus was localized to villus tip enterocytes and gave rise to diarrhoea which was maximal at 72h post-infection (p.i.). Diarrhoea was preceded by ischemia of villi (18-42h p.i.) and villus shortening (maximal at 42h p.i.), and was also coincident with a dramatic regrowth of villi. At 48h p.i. a proliferative zone of electron lucent cells was observed in villus base regions. Cryosections of infected gut, taken before, during, and after infection, together with corresponding age-matched controls, were freeze-dried and analysed by X-ray microanalysis. At 48h p.i. electron lucent villus base cells were shown to be more hydrated, and, to contain higher levels of both Na and Cl and lower levels of P, S, K and Mg than corresponding control cells. These studies increase confidence in the use of X-ray microanalysis in studying biological systems, provide some insight into the process of cell division, and constitute the basis of a new concept of diarrhoeal secretion.27 references.

Stephen, J.; Osborne, M.P.; Spencer, A.J.; Warley, A. (Univ. of Birmingham (England))

1990-09-01

53

From HeLa cell division to infectious diarrhoea  

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Hela S3 cells were grown in suspension both randomly and, synchronously using hydroxyurea which blocks cells at the G1/S interface. Cryosections were prepared, freeze-dried and analyzed by X-ray microanalysis. As cells moved into S and through M phases [Na] and [Cl] increased; both returned to normal levels upon re-entering G1 phase. The Na/K ratio was 1:1 in G1 phase. Infection of HeLa S3 cells in G1 phase with vaccinia virus resulted in no change in intracellular [Na]. Infection of neonatal mice with murine rotavirus was localized to villus tip enterocytes and gave rise to diarrhoea which was maximal at 72h post-infection (p.i.). Diarrhoea was preceded by ischemia of villi (18-42h p.i.) and villus shortening (maximal at 42h p.i.), and was also coincident with a dramatic regrowth of villi. At 48h p.i. a proliferative zone of electron lucent cells was observed in villus base regions. Cryosections of infected gut, taken before, during, and after infection, together with corresponding age-matched controls, were freeze-dried and analysed by X-ray microanalysis. At 48h p.i. electron lucent villus base cells were shown to be more hydrated, and, to contain higher levels of both Na and Cl and lower levels of P, S, K and Mg than corresponding control cells. These studies increase confidence in the use of X-ray microanalysis in studying biological systems, provide some insight into the process of cell division, and constitute the basis of a new concept of diarrhoeal secretion.27 references

1990-01-01

54

Growth inhibition of HeLa cell by internalization of Mycobacterium bovis Bacillus Calmette-Guérin (BCG Tokyo  

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Full Text Available Abstract Background Intravesical BCG immunotherapy is effective for preventing recurrence and progression in none muscle-invasive bladder cancer but the dosing schedule and duration of treatment remain empirical. The mechanisms by which intravesical BCG treatment mediates antitumor activity are currently poorly understood. Results HeLa cell infected with Mycobacterium bovis Bacillus Calmette-Guérin(BCG Tokyo which were different multiplicity of infection(MOI. Proliferation of HeLa cell reduced in a dose-dependent manner by live BCG. The cytoplasm of the HeLa cell showed variety lysosomal stages by internalized and interacted BCG. Conclusion Proliferated Live BCG secreted the protein and depressed the growth of tumor. The possibility for clinical introduction of BCG therapy for carcinoma reported with review of literature.

Asahina Izumi

2009-12-01

55

Growth inhibition of HeLa cell by internalization of Mycobacterium bovis Bacillus Calmette-Gu?rin (BCG) Tokyo  

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Background Intravesical BCG immunotherapy is effective for preventing recurrence and progression in none muscle-invasive bladder cancer but the dosing schedule and duration of treatment remain empirical. The mechanisms by which intravesical BCG treatment mediates antitumor activity are currently poorly understood. Results HeLa cell infected with Mycobacterium bovis Bacillus Calmette-Guérin(BCG) Tokyo which were different multiplicity of infection(MOI). Proliferation of HeLa cell reduced in a dose-dependent manner by live BCG. The cytoplasm of the HeLa cell showed variety lysosomal stages by internalized and interacted BCG. Conclusion Proliferated Live BCG secreted the protein and depressed the growth of tumor. The possibility for clinical introduction of BCG therapy for carcinoma reported with review of literature.

2009-01-01

56

Hemagglutinin protein of measles virus induces apoptosis of HeLa cells via both extrinsic and intrinsic pathways.  

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In this study, we investigated the potential for different components of the measles virus (MV) to induce apoptosis of HeLa cells and explored the apoptotic molecular mechanisms. After testing the 2 envelope glycoproteins hemagglutinin (H) and fusion (F), we found that MV H alone was sufficient to induce the apoptosis of HeLa cells, whereas MV F did not. MV F also had no influence on MV-H-mediated apoptosis. MV H could induce cellular apoptosis in HeLa cells through its interaction with the cellular receptor CD46 via both the TRAIL-mediated extrinsic pathway and the mitochondria-controlled intrinsic pathway, and that cross talk between these 2 pathways occurred during the process. These findings extend the functions of MV envelope glycoproteins in the pathogenesis of MV infection and suggest that MV H may be a potential therapeutic in the treatment of some cancers. PMID:24313454

Yi, Changhua; Liu, Xin; Liu, Yingle; Lu, Songya; Qi, Yipeng

2013-12-01

57

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

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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

2013-10-01

58

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

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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. PMID:23799549

Park, Woo Hyun

2013-08-01

59

Induction of apoptosis in HeLa cells via caspase activation by resveratrol and genistein.  

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Selectively inducing apoptosis in cancer cells is a much desired strategy when tolerance toward side effects is minimal during chemotherapy. In our search for natural products that can induce apoptosis in human cervical cancer cells (HeLa), we selected resveratrol and genistein for our study. We conducted several experiments to test whether genistein can synergistically enhance the apoptotic potential of resveratrol at doses lower than the usual cytotoxic dose. Both resveratrol and genistein were able to induce apoptosis by enhancing the activities of caspase-9 and caspase-3 by themselves and also in combination. After 24?h of exposure to resveratrol and genistein, individually or in combination, lowered mitochondrial membrane potential was observed in HeLa cells. In addition, the mitochondrial membrane potential in HeLa cells was decreased, forcing JC-1 to stay in the monomeric form. The monomeric JC-1(5,5',6,6' -tetrachloro-1,1',3,3'-tetraethyl benzimedazolyl carbocyanine iodide) emitted green fluorescence. In the control group, the color of the fluorescence was red due to aggregation of JC-1 in the physiological pH. The treatment groups exhibited DNA fragmentation as the hallmark of apoptotic nuclear features. We also detected an obvious decrease in the level of HDM2 gene expression after both individual and combination treatments with resveratrol and genistein. Our findings suggest that resveratrol and genistein when combined can induce apoptosis at doses lower than usual doses, through the activation of caspases cascade, and by decreasing the expression of HDM2. PMID:23356442

Dhandayuthapani, Sivanesan; Marimuthu, Palanisamy; Hörmann, Vanessa; Kumi-Diaka, James; Rathinavelu, Appu

2013-02-01

60

In vitro polymerization of microtubules from HeLa cells.  

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Although the purification of microtubules from brain by alternate cycles of polymerization and depolymerization in vitro has become routine, the application of this method to non-neural cultured cells has been less successful. Previous investigations have suggested that it was necessary to use substrate-grown cells and 4 M glycerol to obtain microtubules from cultured cells. We have developed a method for preparing microtubules from HeLa cells in spinner cultures without the use of glycerol. Microtubules can be readily carried through two complete cycles of polymerization at 37 degrees C and depolymerization at 4 degrees C in vitro. The microtubules obtained are morphologically similar to brain microtubules in electron micrographs, and the tubulin subunits have mobilities similar to those of brain tubulins on polyacrylamide gels. Typical yields in the second polymerization pellet are about 1 mg protein/ml of packed cells or 2.5-3.0% of the total protein in the soluble cell extract. The major nontubulin protein present after two cycles of polymerization and depolymerization has an apparent mol wt of 68,000 daltons. If glycerol is used during polymerization, this band is virtually absent. PMID:670297

Weatherbee, J A; Luftig, R B; Weihing, R R

1978-07-01

 
 
 
 
61

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

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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

62

Overexpression of atypical protein kinase C in HeLa cells facilitates macropinocytosis via Src activation.  

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Atypical protein kinase C (aPKC) is the first recognized kinase oncogene. However, the specific contribution of aPKC to cancer progression is unclear. The pseudosubstrate domain of aPKC is different from the other PKC family members, and therefore a synthetic peptide corresponding to the aPKC pseudosubstrate (aPKC-PS) sequence, which specifically blocks aPKC kinase activity, is a valuable tool to assess the role of aPKC in various cellular processes. Here, we learned that HeLa cells incubated with membrane permeable aPKC-PS peptide displayed dilated heterogeneous vesicles labeled with peptide that were subsequently identified as macropinosomes. A quantitative membrane binding assay revealed that aPKC-PS peptide stimulated aPKC recruitment to membranes and activated Src. Similarly, aPKC overexpression in transfected HeLa cells activated Src and induced macropinosome formation. Src-aPKC interaction was essential; substitution of the proline residues in aPKC that associate with the Src-SH3 binding domain rendered the mutant kinase unable to induce macropinocytosis in transfected cells. We propose that aPKC overexpression is a contributing factor to cell transformation by interacting with and consequently promoting Src activation and constitutive macropinocytosis, which increases uptake of extracellular factors, required for altered cell growth and accelerated cell migration. PMID:24582589

Tisdale, Ellen J; Shisheva, Assia; Artalejo, Cristina R

2014-06-01

63

Alteration in the radiosensitivity of HeLa cells by dichloromethane extract of guduchi (Tinospora cordifolia).  

Science.gov (United States)

Exposure of HeLa cells to TCE (dichloromethane extract of Tinospora cordifolia) for 4 hours before exposure to 2-Gy ?-radiation caused a significant decrease in the cell viability (approximately 50%). The surviving fraction (SF) was reduced to 0.52 after 4 hours of TCE treatment; thereafter, clonogenecity of HeLa cells declined negligibly with treatment duration up to 6 hours posttreatment. Exposure of HeLa cells to different doses of ?-radiation resulted in a dose-dependent decline in the viability of HeLa cells, whereas treatment of HeLa cells with various doses of TCE further decreased the cell viability depending not only on the irradiation dose but also on the concentration of TCE. Treatment of HeLa cells with various doses of TCE caused a significant decline in cell viability after exposure to 1 to 4 Gy ?-radiation. The increase in TCE concentration before irradiation caused a concentration-dependent reduction in the SF, and a lowest SF was observed for 4 ?g/mL TCE for all exposure doses. HeLa cells treated with TCE showed an increase in lactate dehydrogenase and decrease in glutathioneS-transferase activity at all postirradiation times. Lipid peroxidation increased up to 4 hours postirradiation and declined gradually up to 12 hours postirradiation. PMID:21106617

Rao, Shaival K; Rao, Priya S

2010-12-01

64

Expression of P-glycoprotein in HeLa cells confers resistance to ceramide cytotoxicity.  

Science.gov (United States)

The role of glucosylceramide synthase (GCS) in regulating ceramide-induced apoptosis has been widely studied. The purpose of this investigation was to evaluate the role of P-glycoprotein (P-gp) in regulating ceramide cytotoxicity by using C6-ceramide. To accomplish this, we employed HeLa cells with conditional expression of the multidrug resistance gene 1/P-gp. HeLa cells expressing P-gp (P-gp/on cells) challenged with [14C]C6-ceramide (6 µM), synthesized 4.5-fold the amount of C6-glucosylceramide (GC) compared to HeLa cells with suppressed expression of P-gp (P-gp/off cells), whereas the generated levels of C6-sphingomyelin were almost equal (33 and 29% of intracellular 14C, respectively). Tamoxifen, a P-gp antagonist, decreased the C6-GC levels from 3.5-1.0% in the P-gp/off and from 17-2.8% of the total lipid 14C levels in the P-gp/on cells. Tamoxifen did not inhibit cell-free C6-GC synthesis in the P-gp/off or P-gp/on homogenates. However, a specific GCS inhibitor, ethylenedioxy-1-phenyl-2-hexadecanoylamino-3-pyrrolidino-1-propanol (ethylenedioxy-P4), blocked synthesis by 90%. In the cytotoxicity assays, the P-gp/off cells were sensitive to C6-ceramide and the P-gp/on cells were resistant. Resistance to C6-ceramide in the P-gp/on cells was reversed by tamoxifen but not by ethylenedioxy-P4. Experiments in another cervical cancer model showed that multidrug-resistant P-gp-rich KB-V1 cells synthesized 3-fold more C6-GC from C6-ceramide than the parental, P-gp-poor KB-3-1 cells, and whereas tamoxifen had no effect on the C6-GC synthesis in the KB-3-1 cells, it inhibited synthesis by 70% in the KB-V1 cells. This study demonstrates that P-gp potentiates C6-ceramide glycosylation and if antagonized augments C6-ceramide sensitivity, both features previously ascribed to GCS. We propose that P-gp can be an effective target for enhancing short-chain ceramide cytotoxicity in the treatment of drug-resistant cancer. PMID:21042729

Chapman, Jacqueline V; Gouazé-Andersson, Valérie; Cabot, Myles C

2010-12-01

65

UDP-Glucuronosyltransferase (UGT) 1A9-Overexpressing HeLa Cells Is an Appropriate Tool to Delineate the Kinetic Interplay between Breast Cancer Resistance Protein (BRCP) and UGT and to Rapidly Identify the Glucuronide Substrates of BCRP  

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The interplay between phase II enzymes and efflux transporters leads to extensive metabolism and low bioavailability for flavonoids. To investigate the simplest interplay between one UDP-glucuronosyltransferase isoform and one efflux transporter in flavonoid disposition, engineered HeLa cells stably overexpressing UGT1A9 were developed, characterized, and further applied to investigate the metabolism of two model flavonoids (genistein and apigenin) and excretion of their glucuronides. The res...

Jiang, Wen; Xu, Beibei; Wu, Baojian; Yu, Rong; Hu, Ming

2012-01-01

66

Anticancer Activity of Certain Herbs and Spices on the Cervical Epithelial Carcinoma (HeLa) Cell Line  

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Acetone extracts of selected plant species were evaluated for their in vitro cytotoxicity against a noncancerous African green monkey kidney (Vero) cell line and an adenocarcinoma cervical cancer (HeLa) cell line. The plants studied were Origanum vulgare L. (Oregano), Rosmarinus officinalis L. (Upright and ground cove rosemary), Lavandula spica L. (Lavender), Laurus nobilis L. (Bay leaf), Thymus vulgaris L. (Thyme), Lavandula x intermedia L. (Margaret Roberts Lavender), Petroselinum crispum M...

Berrington, Danielle; Lall, Namrita

2012-01-01

67

Microtubule-associated proteins of HeLa cells: heat stability of the 200,000 mol wt HeLa MAPs and detection of the presence of MAP-2 in HeLa cell extracts and cycled microtubules.  

Science.gov (United States)

One of the major groups of microtubule-associated proteins (MAPs) found associated with the microtubules isolated from HeLa cells has a molecular weight of just over 200,000. Previous work has demonstrated that these heLa MAPs are similar in several properties to MAP-2, one of the major MAPs of mammalian neural microtubules, although the two types of proteins are immunologically distinct. The 200,000 mol wt HeLa MAPs have now been found to remain soluble after incubation in a boiling water bath and to retain the ability to promote tubulin polymerization after this treatment, two unusual properties also shown by neural MAP-2. This property of heat stability has allowed the development of a simplified procedure for purification of the 200,000 HeLa MAPs and has provided a means for detection of these proteins, even in crude cell extracts. These studies have also led to the detection of a protein in crude extracts of HeLa cells and in cycled HeLa microtubules which has been identified as MAP-2 on the basis of (a) comigration with calf brain MAP-2 on SDS PAGE, (b) presence in purified microtubules, (c) heat stability, and (d) reaction with two types of antibodies prepared against neural high molecular weight-MAPs, one of these a monoclonal antibody against hog brain MAP-2, although present in HeLa cells, is at all stages of microtubule purification a relatively minor component in comparison to the 200,000 HeLa MAP's. PMID:6173388

Weatherbee, J A; Sherline, P; Mascardo, R N; Izant, J G; Luftig, R B; Weihing, R R

1982-01-01

68

Campylobacter jejuni cell lysates differently target mitochondria and lysosomes on HeLa cells.  

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Campylobacter jejuni is the most common cause of bacterial gastroenteritis in humans. The synthesis of cytolethal distending toxin appears essential in the infection process. In this work we evaluated the sequence of lethal events in HeLa cells exposed to cell lysates of two distinct strains, C. jejuni ATCC 33291 and C. jejuni ISS3. C. jejuni cell lysates (CCLys) were added to HeLa cell monolayers which were analysed to detect DNA content, death features, bcl-2 and p53 status, mitochondria/lysosomes network and finally, CD54 and CD59 alterations, compared to cell lysates of C. jejuni 11168H cdtA mutant. We found mitochondria and lysosomes differently targeted by these bacterial lysates. Death, consistent with apoptosis for C. jejuni ATCC 33291 lysate, occurred in a slow way (>48 h); concomitantly HeLa cells increase their endolysosomal compartment, as a consequence of toxin internalization besides a simultaneous and partial lysosomal destabilization. C. jejuni CCLys induces death in HeLa cells mainly via a caspase-dependent mechanism although a p53 lysosomal pathway (also caspase-independent) seems to appear in addition. In C. jejuni ISS3-treated cells, the p53-mediated oxidative degradation of mitochondrial components seems to be lost, inducing the deepest lysosomal alterations. Furthermore, CD59 considerably decreases, suggesting both a degradation or internalisation pathway. CCLys-treated HeLa cells increase CD54 expression on their surface, because of the action of lysate as its double feature of toxin and bacterial peptide. In conclusion, we revealed that C. jejuni CCLys-treated HeLa cells displayed different features, depending on the particular strain. PMID:24880782

Canonico, B; Campana, R; Luchetti, F; Arcangeletti, M; Betti, M; Cesarini, E; Ciacci, C; Vittoria, E; Galli, L; Papa, S; Baffone, W

2014-08-01

69

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

Directory of Open Access Journals (Sweden)

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.

Alshatwi Ali A

2009-11-01

70

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

71

Leptomycin B increases radiosensitization by trichostain A in HeLa cells  

International Nuclear Information System (INIS)

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

2005-06-01

72

Genistein Inhibition of Topoisomerase II? Expression Participated by Sp1 and Sp3 in HeLa Cell  

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Full Text Available Genistein (4?, 5, 7-trihydroxyisoflavone is an isoflavone compound obtained from plants that has potential applications in cancer therapy. However, the molecular mechanism of the action of genistein on cancer cell apoptosis is not well known. In this study, we investigated the effect of genistein on topoisomerase II-? (Topo II?, an important protein involved in the processes of DNA replication and cell proliferation. The results revealed that inhibition of Topo II? expression through the regulation of Specificity protein 1 and Specificity protein 3 may be one of the reasons for genistein’s induction of HeLa cell apoptosis.

Yunzhi Li

2009-07-01

73

Thioredoxin, glutaredoxin, and thioredoxin reductase from cultured HeLa cells.  

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Thioredoxin and glutaredoxin may be important in regulating cell metabolism by mediating interchanges between sulfhydryl and disulfide groups. Components of the thioredoxin/glutaredoxin system from cultured HeLa cells have been partially purified and characterized by using Escherichia coli adenosine 3'-phosphate 5'-phosphosulfate reductase, a thioredoxin/glutaredoxin-dependent enzyme on the pathway of sulfate reduction, as an assay system. In HeLa cells, a NADPH-thioredoxin reductase and thre...

1981-01-01

74

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

2013-06-01

75

Thioredoxin, glutaredoxin, and thioredoxin reductase from cultured HeLa cells.  

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Thioredoxin and glutaredoxin may be important in regulating cell metabolism by mediating interchanges between sulfhydryl and disulfide groups. Components of the thioredoxin/glutaredoxin system from cultured HeLa cells have been partially purified and characterized by using Escherichia coli adenosine 3'-phosphate 5'-phosphosulfate reductase, a thioredoxin/glutaredoxin-dependent enzyme on the pathway of sulfate reduction, as an assay system. In HeLa cells, a NADPH-thioredoxin reductase and three heat-labile proteins (designated PI, PII, and PIII) that have thioredoxin- or glutaredoxin-like properties are found. Both PI and PIII have molecular masses of approximately 12,000 daltons and are readily reduced by their homologous HeLa thioredoxin reductase. However, only PI can be reduced efficiently by the glutathione system and neither PI nor PIII has inherent glutathione-disulfide oxidoreductase activity. PII has a molecular mass of greater than 30,000 daltons and appears to be associated with a reductase activity. The HeLa NADPH-thioredoxin reductase has been purified to near homogeneity and found to be a 116,000-dalton flavoprotein composed of two 58,000-dalton subunits. The HeLa enzyme has low species and substrate specificity and can reduce HeLa PI and PIII, E. coli thioredoxin and glutaredoxin, and the disulfide bond in 5,5'-dithiobis(2-nitrobenzoic acid). The exact in vivo roles of the HeLa thioredoxin/glutaredoxin system remain to be determined. PMID:6950391

Tsang, M L; Weatherbee, J A

1981-12-01

76

Anti-apoptotic effect of caspase inhibitors on H?O?-treated HeLa cells through early suppression of its oxidative stress.  

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Oxidative stress-induced cytotoxicity in cervical cancer cells may be of toxicological interest. In the present study, the effects of exogenous H2O2 on cell growth and death in HeLa cervical cancer cells were investigated, and the anti-apoptotic effects of various caspase (pan-caspase, caspase-3, -8 or -9) inhibitors on H2O2-treated HeLa cells were also evaluated with regard to reactive oxygen species (ROS) and glutathione (GSH) levels. Based on MTT assays, H2O2 inhibited the growth of HeLa cells with an IC50 value of ~75 µM at 24 h. H2O2 increased the number of dead cells and Annexin V-FITC-positive cells in the HeLa cells, which was accompanied by the activation of caspase-3 and the loss of mitochondrial membrane potential (MMP; ??m). However, relatively higher doses of H2O2 induced necrosis in HeLa cells. Caspase inhibitors significantly prevented H2O2-induced HeLa cell death. H2O2 increased ROS including O2•- at 24 h and increased the activity of catalase in HeLa cells. H2O2 also increased the ROS level at 1 h, and several caspase inhibitors attenuated the increased level at 1 h but not at 6, 12 and 24 h. H2O2 decreased the GSH level in HeLa cells at 1 h, and several caspase inhibitors attenuated the decreased level of GSH at this time. H2O2 induced GSH depletion at 24 h. In conclusion, H2O2 inhibited the growth of HeLa cells via apoptosis and/or necrosis, which was accompanied by intracellular increases in ROS levels and GSH depletion. Caspase inhibitors are suggested to suppress H2O2-induced oxidative stress to rescue HeLa cells at the early time point of 1 h. PMID:24627148

Park, Woo Hyun

2014-05-01

77

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

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Highlights: •Nanosecond pulsed electric field (nsPEF) is a new and unique means for life sciences. •Apoptosis was induced by nsPEF exposure in Jurkat cells. •No signs of apoptosis were detected in HeLa S3 cells exposed to nsPEFs. •Formation of poly(ADP-ribose) was induced in nsPEF-exposed HeLa S3 cells. •Two distinct modes of cell death were activated by nsPEF in a cell-dependent manner. -- Abstract: 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

2013-08-30

78

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

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Highlights: •Nanosecond pulsed electric field (nsPEF) is a new and unique means for life sciences. •Apoptosis was induced by nsPEF exposure in Jurkat cells. •No signs of apoptosis were detected in HeLa S3 cells exposed to nsPEFs. •Formation of poly(ADP-ribose) was induced in nsPEF-exposed HeLa S3 cells. •Two distinct modes of cell death were activated by nsPEF in a cell-dependent manner. -- Abstract: 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, Keiko; Akiyama, Hidenori [Institute of Pulsed Power Science, Kumamoto University, Kumamoto 860-8555 (Japan); Yano, Ken-ichi, E-mail: yanoken@kumamoto-u.ac.jp [Priority Organization for Innovation and Excellence, Kumamoto University, Kumamoto 860-8555 (Japan)

2013-08-30

79

Trypanosoma cruzi trypomastigotes induce cytoskeleton modifications during HeLa cell invasion  

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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 trypoma...

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

2011-01-01

80

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

2013-04-01

 
 
 
 
81

Inhibitory Activity of Synthesized Acetylated Procyanidin B1 Analogs against HeLa S3 Cells Proliferation  

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Full Text Available Proanthocyanidins, also known as condensed tannins and/or oligomeric flavonoids, occur in many edible plants and have various interesting biological activities. Previously, we reported a synthetic method for the preparation of various procyanidins in pure form and described their biological activities. Here, we describe the synthesis of procyanidin B1 acetylated analogs and discuss their inhibition activities against HeLa S3 cell proliferation. Surprisingly, the lower-unit acetylated procyanidin B1 strongly inhibited the proliferation of HeLa S3 cells. This molecule showed much stronger inhibitory activity than did epigallocatechin-3-O-gallate (EGCG, green tea polyphenol, and dimeric compounds that included EGCG as a unit. This result suggests that the phenolic hydroxyl groups of the upper-units in flavan-3-ols are important for their inhibitory activity against cancer cell proliferation and that a hydrophobic lower unit dimer enhances this activity.

Syuhei Okamoto

2014-02-01

82

Inhibitory activity of synthesized acetylated Procyanidin B1 analogs against HeLa S3 cells proliferation.  

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Proanthocyanidins, also known as condensed tannins and/or oligomeric flavonoids, occur in many edible plants and have various interesting biological activities. Previously, we reported a synthetic method for the preparation of various procyanidins in pure form and described their biological activities. Here, we describe the synthesis of procyanidin B1 acetylated analogs and discuss their inhibition activities against HeLa S3 cell proliferation. Surprisingly, the lower-unit acetylated procyanidin B1 strongly inhibited the proliferation of HeLa S3 cells. This molecule showed much stronger inhibitory activity than did epigallocatechin-3-O-gallate (EGCG), green tea polyphenol, and dimeric compounds that included EGCG as a unit. This result suggests that the phenolic hydroxyl groups of the upper-units in flavan-3-ols are important for their inhibitory activity against cancer cell proliferation and that a hydrophobic lower unit dimer enhances this activity. PMID:24500007

Okamoto, Syuhei; Ishihara, Sayaka; Okamoto, Taisuke; Doi, Syoma; Harui, Kota; Higashino, Yusuke; Kawasaki, Takashi; Nakajima, Noriyuki; Saito, Akiko

2014-01-01

83

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.

Hugo Alexandre Oliveira Rocha

2011-04-01

84

B cell translocation gene 2 enhances susceptibility of HeLa cells to doxorubicin-induced oxidative damage.  

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BTG2/TIS21/PC3 (B cell translocation gene 2) has been known as a p53 target gene and functions as a tumor suppressor in carcinogenesis of thymus, prostate, kidney, and liver. Although it has been known that the expression of BTG2/TIS21/PC3 is induced during chemotherapy-mediated apoptosis in cancer cells, a role of BTG2/TIS21/PC3 in cell death remains to be elucidated. In this study, the mechanism and role of BTG2 involved in the enhancement of doxorubicin (DOXO)-induced cell death were examined. Treatment of HeLa cells with DOXO revealed apoptotic phenomena, such as chromatin condensation and cleavage of poly(ADP-ribose) polymerase and lamin A/C with concomitant increase of BTG2/TIS21/PC3 expression. Employing infections of Ad-TIS21 virus and lentivirus with short hairpin RNA to BTG2, the effect of BTG2/TIS21/PC3 on the DOXO-induced apoptosis of HeLa cells and liver cancer cells was evaluated. Not only short hairpin RNA-BTG2 but also N-acetyl-L-cysteine significantly reduced the DOXO-induced HeLa cell death and generation of H2O2. Moreover, forced expression of BTG2/TIS21/PC3 using adenoviral vector augmented DOXO-induced cancer cell death concomitantly with increase of manganese-superoxide dismutase but not catalase, CuZnSOD, and glutathione peroxidase 1. The increased apoptosis by forced expression of BTG2/TIS21/PC3 could be inhibited by N-acetyl-L-cysteine and polyethylene glycol-catalase. These results therefore suggest that BTG2/TIS21/PC3 works as an enhancer of DOXO-induced cell death via accumulation of H2O2 by up-regulating manganese-superoxide dismutase without any other antioxidant enzymes. In summary, BTG2/TIS21/PC3 enhances cancer cell death by accumulating H2O2 via imbalance of the antioxidant enzymes in response to chemotherapy. PMID:18840609

Lim, Young-Bin; Park, Tae Jun; Lim, In Kyoung

2008-11-28

85

Effect of Smac gene on apoptosis of HeLa cells induced by ?-rays  

International Nuclear Information System (INIS)

To explore the effect of Smac gene on apoptosis of HeLa cells induced by ?-ray and its possible mechanisms, the full length cDNA of Smac gene was transferred into HeLa cells. 24 h after transferring, the results of Western Blot indicated the expression of Smac was increased but the expression of Survivin decreased. After HeLa cells was irradiated by ?-rays, Smac gene transferred HeLa/Smac cells showed more cell apoptosis rates and the higher activity of Caspase-3 than vector transferred control HeLa/pcDNA3.1 cells. However, the damage and repair of DNA and the cell cycle don't change significantly, comparing HeLa/Smac cells with HeLa/pcDNA3.1 cells. (authors)

2007-09-01

86

Hypoxia-inducible factor-1? enhances the malignant phenotype of multicellular spheroid HeLa cells in vitro  

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The purpose of this study was to clarify the direct effect of hypoxia-inducible factor-1? (HIF-1?) on tumor growth, apoptosis and migration in vitro. To achieve this aim, a comparison was made of the differences in growth rates, apoptotic indices and cell invasive ability in the human cervical cancer cell line HeLa and the HIF-1?-blocked counterpart in a three-dimensional spheroid culture. A significant decrease in cell proliferation and invasion, and an increase in cell apoptosis were obs...

2010-01-01

87

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

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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 G1/S cell cycle arrest and this may be associated with the RB/E2F1 pathway. However, G1/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, XIAOQING; WU, LINGJIAO; LIU, ZHUO; XIE, LIPING; WANG, SHUO

2013-01-01

88

Xanthorrhizol induces apoptosis via the up-regulation of bax and p53 in HeLa cells.  

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Xanthorrhizol is a sesquiterpenoid compound extracted from Curcuma xanthorrhiza, which is known locally as Temulawak. Traditionally, C. xanthorrhiza was found to have antibacterial, anticancer and anti-inflammatory activity. The rhizome has also been used to treat inflammation in postpartum uterine bleeding. An antiproliferative assay using methylene blue staining revealed that xanthorrhizol inhibited the proliferation of the cervical cancer cell line HeLa with an EC50 value of 6.16 microg/ml. Xanthorrhizol significantly increased apoptosis in HeLa cells, as evaluated by the Tdt-mediated dUTP nick end-labelling (TUNEL) assay and nuclear morphology by Hoechst 33258 staining. Western blot analysis, which was further confirmed by the immunostaining results, implied an up-regulation of tumor suppressor protein p53 and the pro-apoptotic protein Bax, following the treatment with xanthorrhizol. Xanthorrhizol, however, did not affect the expression of the anti-apoptotic protein, Bcl-2 and the viral oncoprotein, E6. Hence, xanthorrhizol is a promising antiproliferative and anticancer agent which induces p53 and Bax-dependent apoptosis in HeLa cervical cancer cells. PMID:16158967

Ismail, Norzila; Pihie, Azimahtol Hawariah Lope; Nallapan, Meenakshii

2005-01-01

89

Toxicity of cadmium sulfide (CdS) nanoparticles against Escherichia coli and HeLa cells.  

Science.gov (United States)

The present study endeavours to assess the toxic effect of synthesized CdS nanoparticles (NPs) on Escherichia coli and HeLa cells. The CdS NPs were characterized by DLS, XRD, TEM and AFM studies and the average size of NPs was revealed as ?3 nm. On CdS NPs exposure bacterial cells changed morphological features to filamentous form and damage of the cell surface was found by AFM study. The expression of two conserved cell division components namely ftsZ and ftsQ in E. coli was decreased both at transcriptional and translational levels upon CdS NPs exposure. CdS NPs inhibited proper cell septum formation without affecting the nucleoid segregation. Viability of HeLa cells declined with increasing concentration of CdS NPs and the IC?? value was found to be 4 ?g/mL. NPs treated HeLa cells showed changed morphology with condensed and fragmented nuclei. Increased level of reactive oxygen species (ROS) was found both in E. coli and HeLa cells on CdS NPs exposure. The inverse correlation between declined cell viabilities and elevated ROS level suggested that oxidative stress seems to be the key event by which NPs induce toxicity both in E. coli and HeLa cells. PMID:23892173

Hossain, Sk Tofajjen; Mukherjee, Samir Kumar

2013-09-15

90

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

91

Toxicity of cadmium sulfide (CdS) nanoparticles against Escherichia coli and HeLa cells  

International Nuclear Information System (INIS)

Highlights: • Toxic effect of CdS NPs on the growth and cell division in E. coli was studied. • CdS NPs affected cell surface topology and cell division. • Downregulation of both FtsZ and FtsQ was observed due to NPs exposure. • CdS NPs affected HeLa cell morphology with fragmented nuclei. • All such effects might be due to elevated oxidative stress. -- Abstract: The present study endeavours to assess the toxic effect of synthesized CdS nanoparticles (NPs) on Escherichia coli and HeLa cells. The CdS NPs were characterized by DLS, XRD, TEM and AFM studies and the average size of NPs was revealed as ?3 nm. On CdS NPs exposure bacterial cells changed morphological features to filamentous form and damage of the cell surface was found by AFM study. The expression of two conserved cell division components namely ftsZ and ftsQ in E. coli was decreased both at transcriptional and translational levels upon CdS NPs exposure. CdS NPs inhibited proper cell septum formation without affecting the nucleoid segregation. Viability of HeLa cells declined with increasing concentration of CdS NPs and the IC50 value was found to be 4 ?g/mL. NPs treated HeLa cells showed changed morphology with condensed and fragmented nuclei. Increased level of reactive oxygen species (ROS) was found both in E. coli and HeLa cells on CdS NPs exposure. The inverse correlation between declined cell viabilities and elevated ROS level suggested that oxidative stress seems to be the key event by which NPs induce toxicity both in E. coli and HeLa cells

2013-09-15

92

Toxicity of cadmium sulfide (CdS) nanoparticles against Escherichia coli and HeLa cells  

Energy Technology Data Exchange (ETDEWEB)

Highlights: • Toxic effect of CdS NPs on the growth and cell division in E. coli was studied. • CdS NPs affected cell surface topology and cell division. • Downregulation of both FtsZ and FtsQ was observed due to NPs exposure. • CdS NPs affected HeLa cell morphology with fragmented nuclei. • All such effects might be due to elevated oxidative stress. -- Abstract: The present study endeavours to assess the toxic effect of synthesized CdS nanoparticles (NPs) on Escherichia coli and HeLa cells. The CdS NPs were characterized by DLS, XRD, TEM and AFM studies and the average size of NPs was revealed as ?3 nm. On CdS NPs exposure bacterial cells changed morphological features to filamentous form and damage of the cell surface was found by AFM study. The expression of two conserved cell division components namely ftsZ and ftsQ in E. coli was decreased both at transcriptional and translational levels upon CdS NPs exposure. CdS NPs inhibited proper cell septum formation without affecting the nucleoid segregation. Viability of HeLa cells declined with increasing concentration of CdS NPs and the IC{sub 50} value was found to be 4 ?g/mL. NPs treated HeLa cells showed changed morphology with condensed and fragmented nuclei. Increased level of reactive oxygen species (ROS) was found both in E. coli and HeLa cells on CdS NPs exposure. The inverse correlation between declined cell viabilities and elevated ROS level suggested that oxidative stress seems to be the key event by which NPs induce toxicity both in E. coli and HeLa cells.

Hossain, Sk Tofajjen; Mukherjee, Samir Kumar, E-mail: dr.samirmukherjee@gmail.com

2013-09-15

93

Intracellular Water Specific MR of Microbead-adherent Cells: HeLa Cell Intracellular Water Diffusion  

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The 1H MR signal arising from flowing extracellular media in a perfused, microbead-adherent cultured cell system can be suppressed with a slice-selective, spin-echo pulse sequence. The signal from intracellular water can, thus, be selectively monitored. Herein, this technique was combined with pulsed field gradients to quantify intracellular water diffusion in HeLa cells. The intracellular water MR diffusion-signal attenuation at various diffusion times was well described by a biophysical mod...

Zhao, L.; Sukstanskii, A. L.; Kroenke, C. D.; Song, J.; Piwnica-worms, D.; Ackerman, J. J. H.; Neil, J. J.

2008-01-01

94

Effect of HA14-1 on apoptosis-regulating proteins in HeLa cells.  

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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 this 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 acids 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 downregulating the effect of anti-apoptotic proteins suggesting that HA14-1 may provide therapeutic potential for the treatment of human cervical cancer. PMID:24118733

Rehman, Kanwal; Tariq, Muhammad; Akash, Muhammad S H; Gillani, Zeeshan; Qazi, Mehmood H

2014-03-01

95

The de novo centriole assembly pathway in HeLa cells: cell cycle progression and centriole assembly/maturation  

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It has been reported that nontransformed mammalian cells become arrested during G1 in the absence of centrioles (Hinchcliffe, E., F. Miller, M. Cham, A. Khodjakov, and G. Sluder. 2001. Science. 291:1547–1550). Here, we show that removal of resident centrioles (by laser ablation or needle microsurgery) does not impede cell cycle progression in HeLa cells. HeLa cells born without centrosomes, later, assemble a variable number of centrioles de novo. Centriole assembly begins with the formation...

2005-01-01

96

Dichloroacetate shifts the metabolism from glycolysis to glucose oxidation and exhibits synergistic growth inhibition with cisplatin in HeLa cells.  

Science.gov (United States)

The unique bioenergetic feature of cancer, aerobic glycolysis or the Warburg effect, is an attractive therapeutic target for cancer therapy. Reversing the glycolytic phenotype may trigger apoptosis in tumor cells. Recently, dichloroacetate (DCA) was proven to produce significant cytotoxic effects in certain tumor cells through this distinct mechanism. In this study, the effect of DCA on the metabolism of cervical cancer HeLa cells was explored and its synergistic growth inhibition with cisplatin was also evaluated. The intracellular changes in HeLa cells following DCA exposure were analyzed through cell viability, intracellular H2O2 and pH levels, mitochondrial membrane potential (MMP), expression of apoptotic proteins and Kv1.5 channel, and intracellular-free Ca2+ concentration ([Ca2+]i). For the evaluation of combination chemotherapy, HeLa cells were treated with a combination of DCA and cisplatin at various concentrations for 48 h. Cell viability was determined by CCK-8 assay and the synergy of the two agents was evaluated using the R index method. DCA shifted the metabolism of HeLa cells from aerobic glycolysis to glucose oxidation as shown by the increased intracellular H2O2 and pH levels. The change of the metabolism modality led to a drop in MMP and the increase of apoptotic proteins (caspase 3 and 9). The increased Kv1.5 expression and decreased [Ca2+]i established a positive feedback loop that resulted in reduced tonic inhibition of caspases. Combination chemotherapy of DCA and cisplatin exhibited a significant synergy in inhibiting the proliferation of HeLa cells. The specific apoptotic mechanism of DCA as distinguished from the cisplatin may be partly responsible for the synergy and further in vivo study on combination chemotherapy of the two agents in cervical cancer xenografts in mice is warranted. PMID:21132264

Xie, Jing; Wang, Bing-Shun; Yu, De-Hong; Lu, Qin; Ma, Jian; Qi, Hong; Fang, Chao; Chen, Hong-Zhuan

2011-02-01

97

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

98

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

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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)...

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

2010-01-01

99

Biophysical characterization of gap-junction channels in HeLa cells  

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HeLa cells seem not to be junctionally coupled when probed with techniques such as Lucifer yellow spreading and/or ionic coupling measured with three inserted microelectrodes. When investigated with double whole-cell patch-clamp measurements, HeLa cells in monolayer cultures were electrically coupled in 39% of the cases with very low transjunctional conductances (average one to five open channels). These gapjunction channels had a single-channel conductance ?=26±6 pS and were voltage-ga...

Eckert, Reiner; Dunina-barkovskaja, Antonina; Hu?lser, Dieter F.

1993-01-01

100

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

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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....

Lu, Xiaoqing; Wu, Lingjiao; Liu, Zhuo; Xie, Liping; Wang, Shuo

2013-01-01

 
 
 
 
101

The evidence of HeLa cell apoptosis induced with tetraethylammonium using proteomics and various analytical methods.  

Science.gov (United States)

Tetraethylammonium (TEA) is a potassium channel (KCh) blocker applied in the functional and pharmacological studies of the KChs. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, a colorimetric assay to quantitatively measure living cells, demonstrated that TEA reduced the HeLa cell viability dose-dependently. Flow cytometry analysis indicated an increased apoptosis rate of the HeLa cell after exposing to TEA. The patch clamp technique revealed that the K(+) current of the HeLa cell was inhibited up to 80% when exposed to TEA. In addition, quantitative real-time PCR approach set up cross-talk among the cytotoxicity of TEA, 4-aminopyridine, and anti-cancer drug such as cisplatin. Using comparative proteomics combined with MALDI-TOF MS/MS, 33 significantly changed proteins were found from TEA treatment group; among these proteins, 12 were up-regulated, and 21 were down-regulated. Here we indicated that these proteins were closely connected with many biological functions such as oxidative stress response, signal transduction, metabolism, protein synthesis, and degradation. Both Western blotting and quantitative real-time PCR approaches further verified these differential proteins. Ingenuity Pathways Analysis software, a tool to analyze "omics" data and model biological system, was applied to analyze the interaction pathways of these proteins. The subcellular locations of the differential proteins are also predicted from Uniprot. All results above can help in our understanding of the mechanism of TEA-induced cytotoxicity and provide potential cancer biomarkers. Various experimental results in this study (like those for cisplatin) indicated that TEA is not only a KCh blocker but also a potential anti-cancer drug. PMID:24297172

Huang, Lin; Huang, Qing-Yu; Huang, He-Qing

2014-01-24

102

Single-walled carbon nanotube interactions with HeLa cells.  

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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 MitoSOXtrade mark 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. PMID:17956629

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

2007-01-01

103

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.

Musselman Inga H

2007-10-01

104

Hypochlorous acid-induced modulation of cellular redox status in HeLa cells.  

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Myeloperoxidase catalyzes the formation of hypochlorous acid (HOCI) via reaction of H2O2 with CI(-) ions. Although HOCI plays a major role in the human immune system by killing bacteria and other invading pathogens, excessive generation of this oxidant causes damage to tissues. Exposure of HeLa cells to HOCI decreased viability, inactivated antioxidant enzymes, damaged mitochondria, and modulated cellular redox status. HOCI also induced significant increases in cellular oxidative damage reflected by lipid peroxidation, protein oxidation, and DNA damage. HOCI-mediated oxidative damage to HeLa cells may perturb the cellular antioxidant defense mechanisms and subsequently lead to a pro-oxidant state. PMID:18704334

Park, Sin Young; Shin, Seoung Woo; Lee, Su-Min; Park, Jeen-Woo

2008-07-01

105

Combined treatment with quercetin and imperatorin as a potent strategy for killing HeLa and Hep-2 cells.  

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The aim of the present study was to assess the effect of quercetin and imperatorin administered separately and in combination on apoptosis and autophagy induction in human cervical carcinoma HeLa cells and laryngeal carcinoma Hep-2 cells cultured in vitro. Conducted MTT measurements proved that quercetin and imperatorin displayed a strong antiproliferative activity manifested in markedly reduction of HeLa and Hep-2 cells viability as a result of treatment with 50 ?M of each compound. Further cell staining assays revealed that concentration mentioned above generated the highest percentage of apoptotic cells especially in the case of application of both drugs for 48 h. Simultaneous quercetin and imperatorin administration induced apoptosis remarkably stronger than treatment with single drugs. Experiments at the molecular level confirmed these results accompanied with the decreased Hsp27 and Hsp72 expression and, in addition, with increased caspases activity. Autophagy was not observed and no significant changes in the expression of beclin-1 were noticed. Additionally, experiments were performed on the above-mentioned cell lines with blocked Hsp27 and Hsp72 expression. In these cells, no significant changes in the sensitivity to apoptosis induction upon quercetin and imperatorin treatment were observed. The present study has provided evidence supporting the potential of the combination of quercetin and imperatorin drugs as a novel tool to be used in anticancer therapy. Our results have also demonstrated that blocking of the Hsp27 and Hsp72 gene expression is not enough to sensitize cancer cells to programmed cell death induction in HeLa and Hep-2 cells. PMID:24682729

B?dziul, Dorota; Jakubowicz-Gil, Joanna; Paduch, Roman; G?owniak, Kazimierz; Gawron, Antoni

2014-07-01

106

Tyrosine phosphorylation of ?-catenin affects its subcellular localization and transcriptional activity of ?-catenin in Hela and Bcap-37 cells.  

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In order to investigate the relationship between tyrosine phosphorylation of ?-catenin and transcriptional activity of ?-catenin in Hela and Bcap-37 cells, genistein (a tyrosine kinase inhibitor) was used to inhibit tyrosine phosphorylation in cells. Our results showed the total ?-catenin protein levels were mainly equal in Hela, Bcap-37 and HK-2 cells, ?-catenin was mainly present in nucleus in Hela and Bcap-37cells, while in HK-2 cell ?-catenin was mainly located in cytoplasm. Genistein could inhibit tyrosine phosphorylation of ?-catenin and downregulate nuclear ?-catenin expression in Hela and Bcap-37 cells. In addition, genistein suppressed Ki-67 promoter activity and Ki-67 protein level, thus promoted cell apoptosis. Furthermore, ?-catenin could increase the Ki-67 promoter activity in Hela and Bcap-37 cells. From these findings we conclude that tyrosine phosphorylation of ?-catenin can regulate the cellular distribution of ?-catenin and affect the transcriptional activity of ?-catenin. PMID:24759800

Qian, He-Ya; Zhang, Ding-Guo; Wang, Hong-Wei; Pei, Dong-Sheng; Zheng, Jun-Nian

2014-06-01

107

Establishment of HeLa cell mutants deficient in sphingolipid-related genes using TALENs.  

Science.gov (United States)

Sphingolipids are essential components in eukaryotes and have various cellular functions. Recent developments in genome-editing technologies have facilitated gene disruption in various organisms and cell lines. We here show the disruption of various sphingolipid metabolic genes in human cervical carcinoma HeLa cells by using transcription activator-like effector nucleases (TALENs). A TALEN pair targeting the human CERT gene (alternative name COL4A3BP) encoding a ceramide transport protein induced a loss-of-function phenotype in more than 60% of HeLa cells even though the cell line has a pseudo-triploid karyotype. We have isolated several loss-of-function mutant clones for CERT, UGCG (encoding glucosylceramide synthase), and B4GalT5 (encoding the major lactosylceramide synthase), and also a CERT/UGCG double-deficient clone. Characterization of these clones supported previous proposals that CERT primarily contributes to the synthesis of SM but not GlcCer, and that B4GalT5 is the major LacCer synthase. These newly established sphingolipid-deficient HeLa cell mutants together with our previously established stable transfectants provide a 'sphingolipid-modified HeLa cell panel,' which will be useful to elucidate the functions of various sphingolipid species against essentially the same genomic background. PMID:24498430

Yamaji, Toshiyuki; Hanada, Kentaro

2014-01-01

108

The study of single anticancer peptides interacting with HeLa cell membranes by single molecule force spectroscopy  

Science.gov (United States)

To determine the effects of biophysical parameters (e.g. charge, hydrophobicity, helicity) of peptides on the mechanism of anticancer activity, we applied a single molecule technique--force spectroscopy based on atomic force microscope (AFM)--to study the interaction force at the single molecule level. The activity of the peptide and analogs against HeLa cells exhibited a strong correlation with the hydrophobicity of peptides. Our results indicated that the action mode between ?-helical peptides and cancer cells was largely hydrophobicity-dependent.To determine the effects of biophysical parameters (e.g. charge, hydrophobicity, helicity) of peptides on the mechanism of anticancer activity, we applied a single molecule technique--force spectroscopy based on atomic force microscope (AFM)--to study the interaction force at the single molecule level. The activity of the peptide and analogs against HeLa cells exhibited a strong correlation with the hydrophobicity of peptides. Our results indicated that the action mode between ?-helical peptides and cancer cells was largely hydrophobicity-dependent. Electronic supplementary information (ESI) available: Peptide design, biophysical properties, biological activities and experimental section. See DOI: 10.1039/c2nr11541g

Shan, Yuping; Huang, Jinfeng; Tan, Juanjuan; Gao, Gui; Liu, Shuheng; Wang, Hongda; Chen, Yuxin

2012-02-01

109

Proteins tightly bound to HeLa cell DNA at nuclear matrix attachment sites.  

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DNA-protein complexes have been isolated from HeLa cell nuclei and nuclear matrix preparations. Two proteins, 55 and 66 kilodaltons in size, remain bound to HeLa DNA after treatment at 80 degrees C in 2% sodium dodecyl sulfate and purification by exclusion chromatography on Sepharose 2B-CL in the presence of 0.3% sodium dodecyl sulfate. These proteins appear to be tightly bound but not covalently linked to the DNA, and they are distributed over the DNA with an average spacing of 40 kilobase p...

Bodnar, J. W.; Jones, C. J.; Coombs, D. H.; Pearson, G. D.; Ward, D. C.

1983-01-01

110

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

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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 deoxynucle...

Zhang, Ting; Du, Jiang; Liu, Liguo; Chen, Xiaolan; Yang, Fang; Jin, Qi

2012-01-01

111

Neutron activation analysis of antimony in chromatin and nucleoids of HeLa cells  

International Nuclear Information System (INIS)

Antimony seems to be cancerogenic in men. In the present investigations we tried to find out if Sb+++ are also bound to the cell nucleus. HeLa cells were incubated with SbCl3 and after a 18 h incubation time cells were lysed and crude chromatin isolated. In this preparation Sb was determined by neutron activation analysis. From the same cell culture nucleoids were prepared by ultracentrifugation and also Sb detected in these structures. 12 refs., 2 tabs. (Author)

1988-01-01

112

Induction of Interferon by the Bour Strain of Trachoma in Hela 229 Cells.  

Science.gov (United States)

An inhibitor with many properties associated with interferon was induced in HeLa 229 cells by the Bour strain of trachoma, a member of the psittacosis-LGV-trachoma group of agents. Interferon induced by Bour was demonstrated by the inhibition of growth of...

H. M. Jenkin Y. K. Lu

1967-01-01

113

A Novel Metabolite from Aspergillus ochraceus JGI 25 Showing Cytotoxicity to Hela Cells.  

Science.gov (United States)

This study aims at the isolation of filamentous fungi, extraction of metabolites, and evaluation of the cytotoxic properties on HeLa cells and normal human lymphocytes. We isolated fungi from the soil by serial dilution method. One of the isolates was chosen and identified as Aspergillus ochraceus Wilhelm (Trichocomaceae) by standard techniques. The metabolites were extracted using methanol. Different concentrations of the extract were evaluated for their potential anticancer activity on HeLa cells by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide assay and the safety of the extract was checked on normal human lymphocytes. The extract was purified by chromatographic techniques like thin-layer chromatography and high-performance liquid chromatography, and subjected to mass spectrometric analysis. The extract showed significant cytotoxic potential on HeLa cells at low concentrations with a half maximal inhibitory concentration value of fungal metabolites or metabolites from other strains of A. ochraceus. The metabolite from A. ochraceus is alkaloid in nature, cytotoxic to HeLa cells, and appears to be a novel with anticancer potentials, which could be explored further for characterization of the active component. PMID:24403650

Nadumane, Varalakshmi K; Venkat, Prerana; Pal, Anamika; Dharod, H; Shukla, Megha; Prashanthi, K

2013-09-01

114

A Novel Metabolite from Aspergillus ochraceus JGI 25 Showing Cytotoxicity to Hela Cells  

Science.gov (United States)

This study aims at the isolation of filamentous fungi, extraction of metabolites, and evaluation of the cytotoxic properties on HeLa cells and normal human lymphocytes. We isolated fungi from the soil by serial dilution method. One of the isolates was chosen and identified as Aspergillus ochraceus Wilhelm (Trichocomaceae) by standard techniques. The metabolites were extracted using methanol. Different concentrations of the extract were evaluated for their potential anticancer activity on HeLa cells by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide assay and the safety of the extract was checked on normal human lymphocytes. The extract was purified by chromatographic techniques like thin-layer chromatography and high-performance liquid chromatography, and subjected to mass spectrometric analysis. The extract showed significant cytotoxic potential on HeLa cells at low concentrations with a half maximal inhibitory concentration value of fungal metabolites or metabolites from other strains of A. ochraceus. The metabolite from A. ochraceus is alkaloid in nature, cytotoxic to HeLa cells, and appears to be a novel with anticancer potentials, which could be explored further for characterization of the active component.

Nadumane, Varalakshmi K.; Venkat, Prerana; Pal, Anamika; Dharod, H.; Shukla, Megha; Prashanthi, K.

2013-01-01

115

Measurement of the cytotoxicity of human lymphocytes using HeLa cells as target  

International Nuclear Information System (INIS)

A simple and convenient method is described for the determination of spontaneous, PHA-induced, and antigen-induced cytotoxic activities of human peripheral blood lymphocytes. It involves measuring ["3H]thymidine incorporation in HeLa cells as target. This methodology presents some advantages over the more commonly used "5"1Cr-release test. (Auth.)

1981-01-30

116

Amplification of sodium- and potassium-activated adenosinetriphosphatase in HeLa cells by ouabain step selection  

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A multistep selection for ouabain resistance was used to isolate a clone of HeLa S3 cells that overproduces the plasma membrane sodium, potassium activated adenosinetriphosphatase (Na+,K+-ATPase). Measurements of specific [3H]ouabain-binding to the resistant clone, C+, and parental HeLa cells indicated that C+ cells contain 8-10 X 10(6) ouabain binding sites per cell compared with 8 X 10(5) per HeLa cell. Plasma membranes isolated from C+ cells by a vesiculation procedure and analyzed for oua...

1984-01-01

117

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

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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 as...

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

2012-01-01

118

Formation of extrachromosomal circular DNA in HeLa cells by nonhomologous recombination.  

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Extrachromosomal circular DNA (eccDNA) generated from chromosomal DNA is found in all mammalian cells and increases with cell stress or aging. Studies of eccDNA structure and mode of formation provide insight into mechanisms of instability of the mammalian genome. Previous studies have suggested that eccDNA is generated through a process involving recombination between repetitive sequences. However, we observed that approximately one half of the small eccDNA fragments cloned from HeLa S3 cell...

Loon, N.; Miller, D.; Murnane, J. P.

1994-01-01

119

Anticancer Activity of Certain Herbs and Spices on the Cervical Epithelial Carcinoma (HeLa) Cell Line.  

Science.gov (United States)

Acetone extracts of selected plant species were evaluated for their in vitro cytotoxicity against a noncancerous African green monkey kidney (Vero) cell line and an adenocarcinoma cervical cancer (HeLa) cell line. The plants studied were Origanum vulgare L. (Oregano), Rosmarinus officinalis L. (Upright and ground cove rosemary), Lavandula spica L. (Lavender), Laurus nobilis L. (Bay leaf), Thymus vulgaris L. (Thyme), Lavandula x intermedia L. (Margaret Roberts Lavender), Petroselinum crispum Mill. (Curly leaved parsley), Foeniculum vulgare Mill. (Fennel), and Capsicum annuum L. (Paprika). Antioxidant activity was determined using a quantitative DPPH (1,1-diphenyl-2-picryl hydrazyl) assay. The rosemary species exhibited effective radical scavenging capacity with 50% inhibitory concentration (IC(50)) of 3.48 ± 0.218??g/mL and 10.84 ± 0.125??g/mL and vitamin C equivalents of 0.351?g and 1.09?g for McConnell's Blue and Tuscan Blue, respectively. Cytotoxicity was measured using XTT (Sodium 3'-[1-(phenyl amino-carbonyl)-3,4-tetrazolium]-bis-[4-methoxy-6-nitro] benzene sulfonic acid hydrate) colorimetric assay. Only L. nobilis and O. vulgare exhibited pronounced effects on the HeLa cell line. Dose-dependent studies revealed IC(50) of 34.46 ± 0.48??g/mL and 126.3 ± 1.00??g/mL on the HeLa cells and on the Vero cells 124.1??g/mL ± 18.26 and 163.8??g/mL ± 2.95 for L. nobilis and O. vulgare, respectively. Light (eosin and haematoxylin staining) and confocal microscopy (Hoechst 33342, acridine orange, and propidium iodide staining) were used to evaluate the cytotoxic mechanism of action for L. nobilis and O. vulgare. PMID:22649474

Berrington, Danielle; Lall, Namrita

2012-01-01

120

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

Science.gov (United States)

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

 
 
 
 
121

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

Science.gov (United States)

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 cytometry 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.

2013-01-01

122

The Study of Cisplatin Effect on Hydrogen Peroxide and pH Level in HeLa Kyoto Cell Line Using Genetically-Encoded Sensors  

Directory of Open Access Journals (Sweden)

Full Text Available The aim of the investigation was to study the changes of hydrogen peroxide level and pH level in cytoplasm of cervical cancer cells HeLa Kyoto on cytotoxic exposure using genetically encoded sensors of hydrogen peroxide and pH. Materials and Methods. In the study we used two cell lines of human cervical cancer HeLa Kyoto containing in cytoplasm a genetically encoded sensor of hydrogen peroxide HyPer2 and a sensor pH HyPer2-C199S. To assess toxic effect of cisplatin on HeLa Kyoto cells we used a standard ??? assay. The changes of pH and hydrogen peroxide (H2O2 level were determined using fluorescence microscopy by modification in proportion between fluorescence intensities at sensor’s excitation at two wavelengths: 500 and 420 nm (F500/F420. During the experiment the cells were kept in incubator at 37.0°C in carbonate-free and serum-free medium ???. Cisplatin solution at final concentration corresponding to IC50 according to ??? assay was added directly in culture medium. MEM with no cisplatin added was used as a control medium. Results. Addition of cisplatin resulted in no changes in hydrogen peroxide and pH level in cytoplasm of HeLa Kyoto cells expressing corresponding sensors during the whole period of observation (20 min. Conclusion. The use of genetically encoded sensors enables to demonstrate cisplatin to have no effect on hydrogen peroxide and pH level in HeLa Kyoto cells.

?.S. Belova

2013-11-01

123

Transport of NaYF4:Er3+, Yb3+ up-converting nanoparticles into HeLa cells.  

Science.gov (United States)

An effective, simple and practically useful method to incorporate fluorescent nanoparticles inside live biological cells was developed. The internalization time and concentration dependence of a frequently used liposomal transfection factor (Lipofectamine 2000) was studied. A user friendly, one-step technique to obtain water and organic solvent soluble Er(3+) and Yb(3+) doped NaYF4 nanoparticles coated with polyvinylpyrrolidone was obtained. Structural analysis of the nanoparticles confirmed the formation of nanocrystals of the desired sizes and spectral properties. The internalization of NaYF4 nanoparticles in HeLa cervical cancer cells was determined at different nanoparticle concentrations and for incubation periods from 3 to 24 h. The images revealed a redistribution of nanoparticles inside the cell, which increases with incubation time and concentration levels, and depends on the presence of the transfection factor. The study identifies, for the first time, factors responsible for an effective endocytosis of the up-converting nanoparticles to HeLa cells. Thus, the method could be applied to investigate a wide range of future 'smart' theranostic agents. Nanoparticles incorporated into the liposomes appear to be very promising fluorescent probes for imaging real-time cellular dynamics. PMID:23669145

Sikora, Bo?ena; Fronc, Krzysztof; Kami?ska, Izabela; Koper, Kamil; Szewczyk, Sebastian; Paterczyk, Bohdan; Wojciechowski, Tomasz; Sobczak, Kamil; Minikayev, Roman; Paszkowicz, Wojciech; St?pie?, Piotr; Elbaum, Danek

2013-06-14

124

Inhibition of protein synthesis in intact HeLa cells by Shigella dysenteriae 1 toxin.  

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Shiga toxin purified to near homogeneity from cell lysates of Shigella dysenteriae 1 inhibited protein and deoxyribonucle acid syntheses in intact HeLa cells. Inhibition was dependent on toxin concentration and time of incubation. A minimal latent period of 30 min was observed with saturating doses of toxin. Ribonucleic acid synthesis, uptake of alpha-aminoisobutyric acid, and maintenance of intracellular K+ concentrations were not affected until well after maximal inhibition of protein and d...

Brown, J. E.; Rothman, S. W.; Doctor, B. P.

1980-01-01

125

Potentiation of radiation lethality in HeLa cells by mild hyperthermia and procaine  

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Increased radiation lethality was observed in HeLa cells when irradiation was followed by two hours of incubation at 41/sup o/C in the presence of 1 mM procaine hydrochloride. The increase of radiation lethality produced by this combination was significantly higher than that produced by hyperthermia alone. Two hours of incubation with procaine at 37/sup o/C had no effect on the survival of irradiation cells.

Djordjevic, B. (Mount Sinai Medical Center, New York); Szechter, A.

1981-11-01

126

Human polynucleotide phosphorylase reduces oxidative RNA damage and protects HeLa cell against oxidative stress.  

Science.gov (United States)

We examined HeLa cell viability and RNA oxidative damage in response to hydrogen peroxide (H2O2) treatment. The level of damaged RNA, measured by the content of 8-hydroxyguanosine (7,8-dihydro-8-oxoguanosine, 8-oxoG), increases depending on H2O2 dosage and is inversely correlated with cell viability. The elevated level of 8-oxoG in RNA decreases after removal of oxidative challenge, suggesting the existence of surveillance mechanism(s) for cleaning up oxidized RNA. Human polynucleotide phosphorylase (hPNPase), an exoribonuclease primarily located in mitochondria, has been previously shown to bind 8-oxoG-RNA with high affinity. The role of hPNPase in HeLa cell under oxidative stress conditions is examined here. Overexpression of hPNPase reduces RNA oxidation and increases cell viability against H2O2 insult. Conversely, hPNPase knockdown decreases viability and increases 8-oxoG level in HeLa cell exposed to H2O2. Our results suggest that hPNPase plays an important role in protecting cells and limiting damaged RNA under oxidative stress. PMID:18501193

Wu, Jinhua; Li, Zhongwei

2008-07-25

127

Selective killing effect of oxytetracycline, propafenone and metamizole on A549 or Hela cells  

Science.gov (United States)

Objective To determine the selective killing effect of oxytetracycline, propafenone and metamizole on A549 or Hela cells. Methods Proliferation assay, lactate dehydrogenase (LDH) assay, apoptosis detecting, flow cytometry and western blot were performed. Results It was found that treatment with propafenone at the concentration of 0.014 g/L or higher for 48 h could induce apoptosis in Hela cells greatly, while it was not observed in oxytetracycline and metamizole at the concentration of 0.20 g/L for 48 h. Oxytetracycline, propafenone and metamizole all displayed evident inhibitory effects on the proliferation of A549 cells. The results of LDH assay demonstrated that the drugs at the test range of concentration did not cause necrosis in the cells. Propafenone could elevate the protein level of P53 effectively (P<0.01). Conclusions Oxytetracycline, propafenone and metamizol (dipyrone) all displayed evident inhibitory effects on the proliferation of A549 cells. Propafenone also displayed evident inhibitory effects on the proliferation of Hela cells.

Feng, Guihua

2013-01-01

128

Substitued (E-b-(benzoylacrylic acids suppressed survival of neoplastic human HeLa cells  

Directory of Open Access Journals (Sweden)

Full Text Available The bacteriostatic activity of some of alkyl substituted (E-b-(benzoylacrylic 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-(benzoylacrylic acids, against human cervix carcinoma, HeLa, cells. Target HeLa cells were continuously treated with increasing concentrations of substituted (E-b-(benzoylacrylic 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-(benzoylacrylic 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.

I. JURANIC

1999-09-01

129

Independent regulation by sodium butyrate of gonadotropin alpha gene expression and cell cycle progression in HeLa cells.  

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Sodium butyrate alters the growth and gene expression of a variety of differentiating and neoplastic cell types. For example, addition of 5 mM butyrate to HeLa cells is reported to both induce gonadotropin alpha subunit biosynthesis and block cell cycling in G1. We have studied these two actions of butyrate on HeLa cells and found that they are regulated in distinct ways. The induction of alpha subunit synthesis was due to an increase in the rate of transcription of the alpha gene. Using sync...

Darnell, R. B.

1984-01-01

130

Surface-associated hsp60 chaperonin of Legionella pneumophila mediates invasion in a HeLa cell model.  

Science.gov (United States)

HeLa cells have been previously used to demonstrate that virulent strains of Legionella pneumophila (but not salt-tolerant avirulent strains) efficiently invade nonphagocytic cells. Hsp60, a member of the GroEL family of chaperonins, is displayed on the surface of virulent L. pneumophila (R. A. Garduño et al., J. Bacteriol. 180:505-513, 1988). Because Hsp60 is largely involved in protein-protein interactions, we investigated its role in adherence-invasion in the HeLa cell model. Hsp60-specific antibodies inhibited the adherence and invasiveness of two virulent L. pneumophila strains in a dose-dependent manner but had no effect on the association of their salt-tolerant avirulent derivatives with HeLa cells. A monospecific anti-OmpS (major outer membrane protein) serum inhibited the association of both virulent and avirulent strains of L. pneumophila to HeLa cells, suggesting that while both Hsp60 and OmpS may mediate bacterial association to HeLa cells, only virulent strains selectively displayed Hsp60 on their surfaces. Furthermore, the surface-associated Hsp60 of virulent bacterial cells was susceptible to the action of trypsin, which rendered the bacteria noninvasive. Additionally, pretreatment of HeLa cells with purified Hsp60 or precoating of the plastic surface where HeLa cells attached with Hsp60 reduced the adherence and invasiveness of the two virulent strains. Finally, recombinant Hsp60 covalently bound to latex beads promoted the early association of beads with HeLa cells by a factor of 20 over bovine serum albumin (BSA)-coated beads and competed with virulent strains for association with HeLa cells. Hsp60-coated beads were internalized in large numbers by HeLa cells and remained in tight endosomes that did not fuse with other vesicles, whereas internalized BSA-coated beads, for which endocytic trafficking is well established, resided in more loose or elongated endosomes. Mature intracellular forms of L. pneumophila, which were up to 100-fold more efficient than agar-grown bacteria at associating with HeLa cells, were enriched for Hsp60 on the bacterial surface, as determined by immunolocalization techniques. Collectively, these results establish a role for surface-exposed Hsp60 in invasion of HeLa cells by L. pneumophila. PMID:9746556

Garduño, R A; Garduño, E; Hoffman, P S

1998-10-01

131

Biocompatibility of various ferrite nanoparticles evaluated by in vitro cytotoxicity assays using HeLa cells  

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Magnetic nanoparticles for thermotherapy must be biocompatible and possess high thermal efficiency as heating elements. The biocompatibility of Fe{sub 3}O{sub 4} (20-30 nm), ZnFe{sub 2}O{sub 4} (15-30 nm) and NiFe{sub 2}O{sub 4} (20-30 nm) nanoparticles was studied using a cytotoxicity colony formation assay and a cell viability assay. The Fe{sub 3}O{sub 4} sample was found to be biocompatible on HeLa cells. While ZnFe{sub 2}O{sub 4} and NiFe{sub 2}O{sub 4} were non-toxic at low concentrations, HeLa cells exhibited cytotoxic effects when exposed to concentrations of 100 {mu}g/ml nanoparticles.

Tomitaka, Asahi [Department of Electrical and Computer Engineering, Yokohama National University, Tokiwadai 79-5, Yokohama, Kanagawa 240-8501 (Japan)], E-mail: d07gd158@ynu.ac.jp; Hirukawa, Atsuo; Yamada, Tsutomu [Department of Electrical and Computer Engineering, Yokohama National University, Tokiwadai 79-5, Yokohama, Kanagawa 240-8501 (Japan); Morishita, Shin [Department of Mechanical Engineering and Materials Science, Yokohama National University, Tokiwadai 79-5, Yokohama, Kanagawa 240-8501 (Japan); Takemura, Yasushi [Department of Electrical and Computer Engineering, Yokohama National University, Tokiwadai 79-5, Yokohama, Kanagawa 240-8501 (Japan)

2009-05-15

132

Selective targeting of green fluorescent nanodiamond conjugates to mitochondria in HeLa cells.  

Science.gov (United States)

Fluorescent cellular biomarkers play a prominent role in biosciences. Most of the available biomarkers have some drawbacks due to either physical and optical or cytotoxic properties. In view of this, we investigated the potential of green fluorescent nanodiamonds as biomarkers in living cells. Nanodiamonds were functionalized by attaching antibodies that target intracellular structures such as actin filaments and mitochondria. Then, the nanodiamond conjugates were transfected into HeLa cells. Transfections were mediated by 4(th)-generation dendrimers, cationic liposomes and protamine sulfate. Using fluorescence microscopy, we confirmed successful transfections of the nanodiamonds into HeLa cells. Nanodiamond fluorescence could be easily differentiated from cellular autofluorescence. Furthermore, nanodiamonds could be targeted selectively to intracellular structures. Therefore, nanodiamonds are a promising tool for intracellular assays. PMID:19504515

Mkandawire, Msaukiranji; Pohl, Andrea; Gubarevich, Tatiana; Lapina, Victoria; Appelhans, Dietmar; Rödel, Gerhard; Pompe, Wolfgang; Schreiber, Jürgen; Opitz, Jörg

2009-10-01

133

Biocompatibility of various ferrite nanoparticles evaluated by in vitro cytotoxicity assays using HeLa cells  

International Nuclear Information System (INIS)

Magnetic nanoparticles for thermotherapy must be biocompatible and possess high thermal efficiency as heating elements. The biocompatibility of Fe3O4 (20-30 nm), ZnFe2O4 (15-30 nm) and NiFe2O4 (20-30 nm) nanoparticles was studied using a cytotoxicity colony formation assay and a cell viability assay. The Fe3O4 sample was found to be biocompatible on HeLa cells. While ZnFe2O4 and NiFe2O4 were non-toxic at low concentrations, HeLa cells exhibited cytotoxic effects when exposed to concentrations of 100 ?g/ml nanoparticles.

2009-05-01

134

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

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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

135

Antibody ligation of CM1 on cisplatin-exposed HeLa cells induces apoptosis through reactive oxygen species-dependent Fas ligand expression.  

Science.gov (United States)

Centrocyte/centroblast marker 1 (CM1) has been identified as a pro-apoptosis molecule on B-cell lymphoma cells as well as several types of cancer cells. In this study, we investigated its signaling mechanism in HeLa cells after treatment with cisplatin in order to potentially identify a new therapeutic target. The CM1 molecule was induced on the surface of cisplatin-exposed HeLa cells. In these cells, ligation of CM1 with anti-CM1 monoclonal antibodies inhibited cell proliferation and produced reactive oxygen species. Fas ligand (FasL) expression was upregulated without upregulating Fas in cisplatin-exposed HeLa cells after CM1 stimulation. Pretreatment with N-acetylcysteine, a pan-capase inhibitor, and ZB4, an antagonistic anti-Fas antibody, effectively inhibited the apoptotic effect triggered by CM1. CM1 ligation induced apoptosis through disruption of the mitochondrial membrane potential, decreased Bcl-2 and phosphorylated ERK expression. These findings identify CM1 as a potential new therapeutic target related to cisplatin-exposed cervical cancer. PMID:24682485

Park, Ga Bin; Kim, Daejin; Yoon, Hoi Soo; Kim, Yeong-Seok; Lee, Hyun-Kyung; Kim, Ki Tae; Jeong, Dae Hoon; Hur, Dae Young

2014-06-01

136

Effect of lectin from the ascidian on the growth and the adhesion of HeLa cells.  

Science.gov (United States)

Cell adhesion molecules, some of which are lectins, play a key role in the control of normal and pathological processes of various living organisms. We found herein that N-acetyl-D-glucosamine-specific lectin, isolated from the ascidian Didemnum ternatanum (DTL), alters the growth properties of HeLa tumor cells depending on the anchorage. DTL was shown to increase the proliferation of HeLa cells grown in soft agar greatly (in anchorage-independent fashion). In contrast, DTL inhibits the proliferative activity of HeLa cells grown on solid substrate and acts as inductor of differentiation, slowing cell growth, increasing the cell attachment and spreading. Scanning electron microscopic data have demonstrated that DTL treatment resulted in pronounced changes of the shape and surface of HeLa cells. Changes of cellular morphology correlated with essential redistribution of actin microfilaments. PMID:11506176

Odintsova, N A; Belogortseva, N I; Khomenko, A V; Chikalovets, I V; Luk'yanov, P A

2001-05-01

137

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

Science.gov (United States)

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. PMID:23986283

Svalastog, Anna Lydia; Martinelli, Lucia

2013-08-01

138

A key inactivation factor of HeLa cell viability by a plasma flow  

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Recently, a plasma flow has been applied to medical treatment using effects of various kinds of stimuli such as chemical species, charged particles, heat, light, shock wave and electric fields. Among them, the chemical species are known to cause an inactivation of cell viability. However, the mechanisms and key factors of this event are not yet clear. In this study, we focused on the effect of H2O2 in plasma-treated culture medium because it is generated in the culture medium and it is also chemically stable compared with free radicals generated by the plasma flow. To elucidate the significance of H2O2, we assessed the differences in the effects of plasma-treated medium and H2O2-added medium against inactivation of HeLa cell viability. These two media showed comparable effects on HeLa cells in terms of the survival ratios, morphological features of damage processes, permeations of H2O2 into the cells, response to H2O2 decomposition by catalase and comprehensive gene expression. The results supported that among chemical species generated in a plasma-treated culture medium, H2O2 is one of the main factors responsible for inactivation of HeLa cell viability. (fast track communication)

2011-09-21

139

A key inactivation factor of HeLa cell viability by a plasma flow  

Science.gov (United States)

Recently, a plasma flow has been applied to medical treatment using effects of various kinds of stimuli such as chemical species, charged particles, heat, light, shock wave and electric fields. Among them, the chemical species are known to cause an inactivation of cell viability. However, the mechanisms and key factors of this event are not yet clear. In this study, we focused on the effect of H2O2 in plasma-treated culture medium because it is generated in the culture medium and it is also chemically stable compared with free radicals generated by the plasma flow. To elucidate the significance of H2O2, we assessed the differences in the effects of plasma-treated medium and H2O2-added medium against inactivation of HeLa cell viability. These two media showed comparable effects on HeLa cells in terms of the survival ratios, morphological features of damage processes, permeations of H2O2 into the cells, response to H2O2 decomposition by catalase and comprehensive gene expression. The results supported that among chemical species generated in a plasma-treated culture medium, H2O2 is one of the main factors responsible for inactivation of HeLa cell viability.

Sato, Takehiko; Yokoyama, Mayo; Johkura, Kohei

2011-09-01

140

Binding of the glycan of the major outer membrane protein of Chlamydia trachomatis to HeLa cells.  

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Recent studies have shown that the major outer membrane protein (MOMP) of Chlamydia trachomatis is glycosylated. The glycan of the MOMP of C. trachomatis serovar L2 was separated from the glycoprotein with N-glycanase, reduced with tritiated NaBH4, and tested for its ability to interact with HeLa cells. The [3H]glycan was shown to attach readily to HeLa cells at 25 or 37 degrees C. This process was slower at 4 degrees C. Competition for possibly similar receptor sites on HeLa cells between th...

Swanson, A. F.; Kuo, C. C.

1994-01-01

 
 
 
 
141

Independent induction of sister-chromatid exchanges by 3-aminobenzamide and ultraviolet radiation in HeLa cells  

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The effect of 3-aminobenzamide in HeLa cells was examined in several aspects. 10-3 M 3-aminobenzamide inhibited poly(ADP-ribose) polymerase activity to more than 95% in disrupted nuclei of HeLa cells. The mode of inhibition of the enzyme was competitive inhibition with NAD at a Ksub(i) value of 2.5 ?M 3-aminobenzamide. The combined treatment of HeLa cells with 3-aminobenzamide and ultraviolet radiation revealed the independent induction of SCEs by these 2 agents. (orig.)

1983-01-01

142

Trypanosoma cruzi trypomastigotes induce cytoskeleton modifications during HeLa cell invasion  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english 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.

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

143

Effect of different stress factors on IL-6 and leptin expression in HELA cell cultures  

International Nuclear Information System (INIS)

Objective: To study the effect of three stress factors high glucose (HG), lipopolysaccharide (LPS) and hydrogen peroxide (H2O2) on the expression of culture supernatant IL-6 (IL-6) and leptin contents of HELA cell line. Methods: HELA cell culture models of severe inflammatory response syndrome were prepared with cultures treated with 50 mmol/L glucose (HG), 4 ?g/ ml LPS and 100 ?mol/L H2O2 respectively and supernatant contents of IL-6 and leptin were measured with RIA at 1h, 6h and 24h. Results: Generally speaking, the culture supernatant contents of IL-6 gradually increased and leptin contents gradually decreased with significant differences from those in cultures not treated with either stress factor at 6h and 12h (P<0.05). Conclusion: Leptin as a possible anti-inflammatory cytokine might plays an important protective role in severe inflammatory response. (authors)

2009-02-01

144

Biosynthesis of gold nanoparticles using Sargassum swartzii and its cytotoxicity effect on HeLa cells  

Science.gov (United States)

In this investigation, biological synthesis of gold nanoparticles (AuNPs) using Sargassum swartzii and its cytotoxicity against human cervical carcinoma (HeLa) cells is reported. The biological synthesis involved the reduction of chloroauric acid led to the formation of AuNPs within 5 min at 60 °C and the formation of AuNPs was confirmed using UV-vis spectrophotometer. The AuNPs were stable; spherical in shape with well-defined dimensions, and the average size of the particle is 35 nm. A zeta potential value of -27.6 mV revealed synthesized AuNPs were highly stable. The synthesized AuNPs exhibited a dose-dependent cytotoxicity against human cervical carcinoma (HeLa) cells. Furthermore, induction of apoptosis was measured by DAPI (4?,6-Diamidino-2-phenylindole dihydrochloride) staining.

Dhas, T. Stalin; Kumar, V. Ganesh; Karthick, V.; Govindaraju, K.; Shankara Narayana, T.

2014-12-01

145

Modification of some biological properties of HeLa cells containing adeno-associated virus DNA integrated into chromosome 17.  

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Parvoviruses are known to interfere with cellular transformation and carcinogenesis. Since infecting adeno-associated virus (AAV) frequently integrates its DNA into the cellular genome, we analyzed whether this integration influences the transformed phenotype of the human tumor cell line HeLa. Analysis of three independent HeLa cell clones with integrated AAV DNA (HA-3x, HA-16, and HA-28) revealed the following phenotypic changes of these cells: (i) reduced growth rate, (ii) increased serum r...

Walz, C.; Schlehofer, J. R.

1992-01-01

146

Binding of vitronectin to opa-expressing Neisseria gonorrhoeae mediates invasion of HeLa cells.  

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Neisseria gonorrhoeae induces local infections in the human genitourinary tract and can disseminate to other organs to cause severe disease. Blood-derived factors present in the genital mucosa have been suggested to facilitate the spread of N. gonorrhoeae in disseminated gonococcal infections. Using gentamicin invasion assays and confocal microscopy, we observed a strong stimulatory effect of fetal calf serum (FCS) on the gonococcal invasion of HeLa cells. FCS-mediated invasion was dependent ...

Go?mez-duarte, O. G.; Dehio, M.; Guzma?n, C. A.; Chhatwal, G. S.; Dehio, C.; Meyer, T. F.

1997-01-01

147

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

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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 ...

2011-01-01

148

Interaction of Escherichia coli producing cytotoxic necrotizing factor with HeLa epithelial cells.  

Science.gov (United States)

Cytotoxic necrotizing factors (CNF) constitute a group a cell-associated proteic toxins of 110-115 kDa produced by some clinical isolates of Escherichia coli from man and animals. Purified CNFs are known to exacerbate actin polymerization in exposed cells, a property that has been ascribed to their ability to modify rho a small GTP-protein involved in the regulation of the cytoskeleton. We speculated that, in spite of their lack of excretion in broth culture supernatants, CNF might be expressed upon direct interaction of organisms with infected cells. To test this hypothesis, we set up a model of interaction using epithelial cell line HeLa and the CNF1-producing strain BM2-1, which is adherent to Hela cells. An interaction of four hour duration triggers the progressive development of a dose-dependent cytopathic effect (CPE) with following characteristics: (1) intense cell enlargement with formation of a dense network of stress fibers, (2) inhibition of cell mitosis due to an irreversible block in G2/M transition phase, (3) nucleus swelling and fragmentation, and (4) cell death starting five days after infection. The three last features clearly differentiate CPE from the effect produced by CNF1 alone. In addition CPE, was not produced by cell-free culture supernatants nor abolished by an antiserum neutralizing CNF1. Tn5::PhoA insertion in the 3' end of cnf1 structural gene abolished CPE, which was not restored by trans complementation with cloned cnf1. These results demonstrate that CNF1-producing E. coli exert a specific pathogenic effect in HeLa cells, which is determined by cnf1 and at least one additional gene, located downstream cnf1. PMID:9192042

De Rycke, J; Nougayrede, J P; Oswald, E; Mazars, P

1997-01-01

149

Sequence dependence of the hyperthermic potentiation of carboplatin-induced cytotoxicity and intracellular platinum accumulation in HeLa cells.  

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We have examined the enhancement of cytotoxic effects of cis-diammine-1,1-cyclobutane dicarboxylate platinum(II) (carboplatin) by hyperthermia in HeLa cells using different regimes of timing and sequence. The results were compared with those obtained with cis-diamminedichloroplatinum(II) (cisplatin). We found that cisplatin simultaneously combined with heat was the most cytotoxic toward HeLa cells of the various timing and sequencing conditions studied. On the other hand, for carboplatin, dru...

1993-01-01

150

Uptake and fate of ouabain bound to HeLa cells. [/sup 3/H tracer study  

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The first step in the pharmacologic interaction of a cardiac glycoside with cells is the binding of the drug to its specific surface receptor, an exteriorly oriented site of the Na/sup +/--K/sup +/ ATPase. The purpose of the study outlined was to follow a glycoside from its initial binding to its ultimate release from a sensitive cell, and in parallel studies to observe the recovery of the cells from glycoside intoxication. The glycoside used is /sup 3/H-ouabain, and the cells are from the S3 clone of HeLa. HeLa cells are used principally because of their intrinsic interest as a human cell type. In addition, having been cloned, they can be grown as a homogeneous population, which greatly simplifies the interpretation of the results. Finally, after pulse treatment with the drug, the cells can be returned to a complete medium with adequate supplies of serum, amino acids, vitamins, and energy sources. In this normal environment, not only can the short term physiologic responses be assessed but, with the growth requirements of the cells fulfilled, any recovery processes dependent on macromolecular-synthesis can also be observed.

Cook, J. S.; Brake, E. T.

1977-01-01

151

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<0.01. The apoptotic changes and apoptosis bodies could be observed under microscopes. The fluorescence staining revealed that 4HPR could induce the cells apoptosis, not necrosis. Typical DNA ladder appeared in 4HPR-treated cells and detected by FCM, the subdiploid nuclear peak appeared on the left of G1 peak and the apoptotic percentage was correlated with 4HPR in a dose and time dependent manner(P<0.01. All these results indicated that 4HPR could inhibit the proliferation of Hela cells and induce the cells apoptosis.

Xiao-Hong Han

2011-06-01

152

Ethanolic Neem (Azadirachta indica) Leaf Extract Prevents Growth of MCF-7 and HeLa Cells and Potentiates the Therapeutic Index of Cisplatin.  

Science.gov (United States)

The present study was designed to gain insight into the antiproliferative activity of ethanolic neem leaves extract (ENLE) alone or in combination with cisplatin by cell viability assay on human breast (MCF-7) and cervical (HeLa) cancer cells. Nuclear morphological examination and cell cycle analysis were performed to determine the mode of cell death. Further, to identify its molecular targets, the expression of genes involved in apoptosis, cell cycle progression, and drug metabolism was analyzed by RT-PCR. Treatment of MCF-7, HeLa, and normal cells with ENLE differentially suppressed the growth of cancer cells in a dose- and time-dependent manner through apoptosis. Additionally, lower dose combinations of ENLE with cisplatin resulted in synergistic growth inhibition of these cells compared to the individual drugs (combination index <1). ENLE significantly modulated the expression of bax, cyclin D1, and cytochrome P450 monooxygenases (CYP 1A1 and CYP 1A2) in a time-dependent manner in these cells. Conclusively, these results emphasize the chemopreventive ability of neem alone or in combination with chemotherapeutic treatment to reduce the cytotoxic effects on normal cells, while potentiating their efficacy at lower doses. Thus, neem may be a prospective therapeutic agent to combat gynecological cancers. PMID:24624140

Sharma, Chhavi; Vas, Andrea J; Goala, Payal; Gheewala, Taher M; Rizvi, Tahir A; Hussain, Arif

2014-01-01

153

Ethanolic Neem (Azadirachta indica) Leaf Extract Prevents Growth of MCF-7 and HeLa Cells and Potentiates the Therapeutic Index of Cisplatin  

Science.gov (United States)

The present study was designed to gain insight into the antiproliferative activity of ethanolic neem leaves extract (ENLE) alone or in combination with cisplatin by cell viability assay on human breast (MCF-7) and cervical (HeLa) cancer cells. Nuclear morphological examination and cell cycle analysis were performed to determine the mode of cell death. Further, to identify its molecular targets, the expression of genes involved in apoptosis, cell cycle progression, and drug metabolism was analyzed by RT-PCR. Treatment of MCF-7, HeLa, and normal cells with ENLE differentially suppressed the growth of cancer cells in a dose- and time-dependent manner through apoptosis. Additionally, lower dose combinations of ENLE with cisplatin resulted in synergistic growth inhibition of these cells compared to the individual drugs (combination index <1). ENLE significantly modulated the expression of bax, cyclin D1, and cytochrome P450 monooxygenases (CYP 1A1 and CYP 1A2) in a time-dependent manner in these cells. Conclusively, these results emphasize the chemopreventive ability of neem alone or in combination with chemotherapeutic treatment to reduce the cytotoxic effects on normal cells, while potentiating their efficacy at lower doses. Thus, neem may be a prospective therapeutic agent to combat gynecological cancers.

Sharma, Chhavi; Vas, Andrea J.; Goala, Payal; Gheewala, Taher M.; Rizvi, Tahir A.

2014-01-01

154

Modulation of bacterial association to HeLa cell cultures by cell density and by chlamydial infection.  

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The effect of cell density on the rate of association of Escherichia coli and Staphylococcus aureus by monolayer cultures of HeLa 229 cells was investigated. Radioactively labeled bacteria were incubated with sparsely and densely plated cells. The rate of bacterial uptake was 10- to 20-fold higher in sparse cultures. Kinetic analysis of data with different multiplicities of input bacteria showed that the Km of the reaction was unaltered, whereas the Vmax was inversely related to cell density....

Bose, S. K.; Mudd, R. L.

1981-01-01

155

Effect of Rta protein of Epstein-Barr virus on the cell cycle in HeLa cells.  

Science.gov (United States)

Epstein-Barr virus (EBV) replication and transcription activator (Rta) is an immediate-early transcription factor that mediates the switch from latent to lytic infection. DNA viruses often modulate the function of critical cell cycle proteins to maximize the efficiency of virus replication. Here we have examined the effect of Rta on cell cycle progression. Cell cycle analysis revealed that Rta induced HeLa cells in G0/G1-phase to reenter the S-phase. Analysis of the expression pattern of a key set of cell cycle regulators revealed that expression of Rta inhibited the expression of Rb and p53 and induced the expression of E2F1. These findings suggest that Rta plays an active role in redirecting HeLa cell physiology through an Rta-mediated cell cycle transformation. PMID:22149496

Guo, Qingwei; Sun, Xiaobai; Yuan, Changjin; Zhou, Huijuan; Li, Yanmei; Jie, Gao; Jiang, Guosheng

2011-01-01

156

The de novo centriole assembly pathway in HeLa cells  

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It has been reported that nontransformed mammalian cells become arrested during G1 in the absence of centrioles (Hinchcliffe, E., F. Miller, M. Cham, A. Khodjakov, and G. Sluder. 2001. Science. 291:1547–1550). Here, we show that removal of resident centrioles (by laser ablation or needle microsurgery) does not impede cell cycle progression in HeLa cells. HeLa cells born without centrosomes, later, assemble a variable number of centrioles de novo. Centriole assembly begins with the formation of small centrin aggregates that appear during the S phase. These, initially amorphous “precentrioles” become morphologically recognizable centrioles before mitosis. De novo–assembled centrioles mature (i.e., gain abilities to organize microtubules and replicate) in the next cell cycle. This maturation is not simply a time-dependent phenomenon, because de novo–formed centrioles do not mature if they are assembled in S phase–arrested cells. By selectively ablating only one centriole at a time, we find that the presence of a single centriole inhibits the assembly of additional centrioles, indicating that centrioles have an activity that suppresses the de novo pathway.

La Terra, Sabrina; English, Christopher N.; Hergert, Polla; McEwen, Bruce F.; Sluder, Greenfield; Khodjakov, Alexey

2005-01-01

157

Phenols of virgin olive oil protects nuclear DNA against oxidative damage in HeLa cells.  

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Oxidative DNA damage is an inescapable consequence for cells constantly exposed to oxidative stress derived from normal metabolic processes and from environmental factors. Phenolic compounds, which have strong antioxidant activity, prevent DNA damage by protecting the cells against harmful effects of oxidative stress. In this study, the effect of virgin olive oil phenolic extract (OOPE) was investigated on H2O2-induced mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) damage in HeLa cells. DNA damage was assessed in mitochondria and two nuclear regions by using quantitative PCR (QPCR) assay. The cells were pre-treated with non-cytotoxic doses of OOPE for 4 h, and DNA damage was determined. OOPE alone does not change the steady-state level of DNA damage. The oxidative stress generated with 750 ?M H2O2 caused two times greater damages in mtDNA compared to nDNA, which included the nonexpressed ?-globin region (1.507±0.110 lesions/10 kb) and the expressed APEX1 gene (1.623±0.243 lesions/10 kb) with respect to the control region. When cells were preincubated with OOPE for 4 h, nDNA damage under stress condition was completely inhibited; however, mtDNA damage was not affected by this procedure. These results suggest that OOPE has a protective effect against nDNA damage in HeLa cells. PMID:22877972

Erol, Özlem; Arda, Nazl?; Erdem, Günhan

2012-10-01

158

Reversal effects of antifungal drugs on multidrug resistance in MDR1-overexpressing HeLa cells.  

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In this study, the antiproliferative effects of vinblastine (VLB), paclitaxel (TXL), doxorubicin (DXR), daunorubicin (DNR) and 5-fluorouracil (5-FU) were assessed in the human cervical carcinoma cell line HeLa-Ohio (HeLa) and Hvr100-6 cells, established by growing the parental HeLa cells in the presence of progressively greater concentrations of VLB in the culture medium. Flow cytometric analysis indicated the induction of MDR1 (P-glycoprotein) in Hvr100-6 cells with no alterations in levels of multidrug resistance-associated protein (MRP). Resistance to VLB, TXL, DXR and DNR was found in Hvr100-6 cells with relative resistances of ca. 300, 4000, 50 and 200, respectively, whereas no resistance was found to 5-FU. The reversal effects of antifungal drugs, fluconazole, itraconazole, ketoconazole, miconazole and amphotericin B on multidrug resistance were also assessed using Hvr100-6 cells. Itraconazole was found to have potent reversal effect on the resistance to VLB and TXL, but the others had no such effect. This reversal effect of itraconazole was concentration-dependent, with dose modifying factors of 3.2, 10.1 and 435.7 at 0.1, 0.25 and 0.5 microM of itraconazole, respectively. In addition, this reversal effect of itraconazole was explained by the inhibition of accumulation of the anticancer drugs. PMID:11558564

Iida, N; Takara, K; Ohmoto, N; Nakamura, T; Kimura, T; Wada, A; Hirai, M; Sakaeda, T; Okumura, K

2001-09-01

159

Effects of several compounds on the chromium uptake from surrounding medium by HeLa cells  

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Radiochromium uptake from surrounding by HeLa cells was examined, the results were as follows: 1) The chromium uptake by the cells after a certain period of incubation in Ca-Mg free phosphate buffered solution (PBS) with radiosodiumchromate (Na251CrO4) was higher than that in serum free Eagle's minimum essential medium with the same concentration of the radiochromate. 2) When the various amount of L-ascorbic acid was added to the above radiochromate containing PBS, the chromium uptake by the cells decreased with dependence on the concentration of the acid in the surrounding medium. However, when sodiumthiosulfate was added to the medium, no remarkable effect was found. 3) When cells were incubated in the radiochromic chloride (51CrCl3) containing medium with 6.5 ?g/ml of sodium oxalate, sodium acetate or sodium nitrate, the chromium uptake by the cells increased in comparison with the control. Above results suggested that the chromium uptake by the HeLa cells from surrounding medium was affected by several chemicals and the uptake or binding capacity of chromium was closely related to the reported cytotoxicity of the chromium compounds. (auth.)

1977-01-01

160

Photoirradiation study of gold nanospheres and rods in Vero and Hela cell lines  

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Photoirradiation effect of gold nanospheres in conjucation with green light and rods in conjugation with red light corresponds to their absorption wavelength range found to be appreciable. In this present work concentration of nanomaterial and light dose were optimized. Gold nanospheres were synthesized by reduction technique using Sodium Borohydrate as reducing agent and Trisodium Citrate as capping agent. Au nanorods having 680-900nm absorption were synthesized using reduction techniques with CTAB and BDAC polymers. From UV-Vis absorption and Transmission Electron Microscopy the size of nanoparticles were confirmed. 30nm Gold nanospheres and green light source of 530nm wavelength with power 30mW were applied to Vero and Hela cell lines shows higher toxicity for Hela cells. Nanorods were applied and irradiated with 680nm wavelength light source with light intensity 45mW. Post irradiation effect for 24hrs, 48hrs confirms cell proliferation in normal rate in viable cells. The morphological changes in irradiated spot leads to apoptotoic cell death was confirmed with microscopic imaging. The LD50 value was also calculated.

Gananathan, Poorani; Aruna, Prakasarao; Ganesan, Singaravelu; Elanchezhiyan, Manickan

2014-03-01

 
 
 
 
161

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

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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

162

Nuclear proteome analysis of cisplatin-treated HeLa cells  

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Cisplatin has been widely accepted as one of the most efficient anticancer drugs for decades. However, the mechanisms for the cytotoxic effects of cisplatin are still not fully understood. Cisplatin primarily targets DNA, resulting in the formation of DNA double strand breaks and eventually causing cell death. In this study, we applied two-dimensional electrophoresis coupled with LC-MS/MS to analyze the nuclear proteome of HeLa cells treated with cisplatin, in an effort to uncover new mechanistic clues regarding the cellular response to cisplatin. A total of 19 proteins were successfully identified, and these proteins are involved in a variety of basal metabolic and biological processes in cells, including biosynthesis, cell cycle, glycolysis and apoptosis. Six were related to the regulation of mRNA splicing, and we therefore asked whether the Fas gene might undergo alternative splicing following cisplatin treatment. This proved to be the case, as the splicing forms of Fas were modified in cisplatin-treated HeLa cells. This work provides novel information, from the perspective of the nuclear response, for understanding the cytotoxicity caused by cisplatin-induced DNA damage.

2010-09-10

163

Structures of nuclear phosphoproteins characteristic of rapidly growing HeLa cells  

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To study characteristic events of phosphorylation in cell growth, phosphoproteins were labeled with [32P]-phosphate at mid-logarithmic phase of HeLa cell proliferation. Among a number of nuclear phosphoproteins isolated, three characteristic classes of most highly labeled phosphoproteins were identified by DEAE-column chromatography (0.2-0.25 M NaCl gradient, pH 6.0), followed by 7.5% SDS polyacrylamide gel electrophoresis. Chemical characterization of their structures showed that they contained three different forms of post-translational modifications: Class I with phosphoserine, Class II with phosphoserine and oligonulceotides (5-10 nucleotides long), and Class III with phosphoserine, 5'-GMP and poly(ADP-ribose). Class I is represented by nucleolar C-23. Class II is represented by nucleolar 125 kDa and nucleoplasmic 50 kDa with GC rich sequences (G = 30%, C = 40%) and 5'-linking pCp. Class III is represented by nucleoplasmic poly(ADP-ribose) proteins (18 different species, MW ranges 30 kDa-200 kDa) with branched poly(ADP-ribose) longer than tRNA. When HeLa cells were labeled at non-mid-logarithmic phase, labeling of these classes were 4 fold less efficient, indicating their functional importance in cell proliferation

1986-05-01

164

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

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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)

2008-12-01

165

Methylation of DNA in HeLa cells after ultraviolet irradiation  

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The synthesis of DNA and its methylation were measured in HeLa cells after exposure to increasing doses of ultraviolet (uv). The extent of methylation relative to DNA synthesis increases in a dose dependent manner. Old and new DNA were spearated by gradient centrifugation and the effect of uv on the methylation of the two species was determined. Methylation of new DNA is affected more extensively than the methylation of old DNA by exposure to irradiation. The possible significant of aberrant methylation of DNA in potentiating uv induced damage is discussed

1976-01-01

166

Methylation of DNA in HeLa cells after ultraviolet irradiation  

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The synthesis of DNA and its methylation were measured in HeLa cells after exposure to increasing doses of ultraviolet (uv). The extent of methylation relative to DNA synthesis increases in a dose dependent manner. Old and new DNA were spearated by gradient centrifugation and the effect of uv on the methylation of the two species was determined. Methylation of new DNA is affected more extensively than the methylation of old DNA by exposure to irradiation. The possible significant of aberrant methylation of DNA in potentiating uv induced damage is discussed.

Low, M.; Read, E.L.; Borek, E.

1976-01-01

167

Effect of powerfulshort laser impulses on permeability of outer membranes of HeLa cells  

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It was shown that in HeLa cells, at the stationary growth phase, the permeability of the outer membrane for thymidine increased 1 h after irradiation with two ultra-shot impulses (Nd3+: YAG laser, ?=266, 532 or 1064 nm, the impulse duration of 3x10-11 s, I=2 or 20 MW/cm2). The effect was dependent on the time interval between the impulses and virtually independent of the radiation wave-length. The maximum increase in the membrane permeability (by ? 3 times) was noted after irradiation at 4-6 s intervals between the impulses

1986-01-01

168

Comparative multiparametric analysis of HeLa and RD cell culture reactions to solcoseryl.  

Science.gov (United States)

Reactions of continuous HeLa and RD cell cultures and their nuclear and nucleolar apparatus to addition of solcoseryl into the medium were studied. The monolayer density, proliferation activity, percentage of dead cells, RNA and DNA content in the nuclei and nucleoli, number of nucleoli in the nuclei, cell distribution in the population by the number of nucleoli in the nuclei, volume and complete surface area of the nuclei and nucleoli, and the nucleolar/nuclear ratio were evaluated. The cultures differently reacted to solcoseryl in the medium at the population and cellular levels of their organization. By the results of multiparametric analysis of the reactions of cells and their nuclear and nucleolar apparatus, solcoseryl can be referred to bioactive substances with characteristics of a factor regulating cell population growth. PMID:20396754

Magakian, Yu A; Karalyan, Z A; Karalova, E M; Abroyan, L O; Akopyan, L A; Gasparyan, M H; Jaghacpanyan, N G; Semerjyan, Z B; Ter-Pogossyan, Z R

2009-10-01

169

Proteomic, cellular, and network analyses reveal new DUSP3 interactions with nucleolar proteins in HeLa cells.  

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DUSP3 (or Vaccinia virus phosphatase VH1-related; VHR) is a small dual-specificity phosphatase known to dephosphorylate c-Jun N-terminal kinases and extracellular signal-regulated kinases. In human cervical cancer cells, DUSP3 is overexpressed, localizes preferentially to the nucleus, and plays a key role in cellular proliferation and senescence triggering. Other DUSP3 functions are still unknown, as illustrated by recent and unpublished results from our group showing that this enzyme mediates DNA damage response or repair processes. In this study, we sought to identify new interactions between DUSP3 and proteins directly or indirectly involved in or correlated with its biological roles in HeLa cells exposed to gamma or UV radiation. By using GST-DUSP as bait, we pulled down interacting proteins and identified them by LC-MS/MS. Of the 46 proteins obtained, six hits were extensively validated by immune techniques; the proteins Nucleophosmin, HnRNP C1/C2, and Nucleolin were the most promising targets found to directly interact with DUSP3. We then analyzed the DUSP3 interactomes using physical protein-protein interaction networks using our hits as the seed list. The validated hits as well as unvalidated hits fluctuated on the DUSP3 interactomes of HeLa cells, independent of the time post radiation, which confirmed our proteomic and experimental data and clearly showed the proximity of DUSP3 to proteins involved in processes intimately related to DNA repair and senescence, such as Ku70 and Tert, via interactions with nucleolar proteins, which were identified in this study, that regulate DNA/RNA structure and functions. PMID:24245651

Panico, Karine; Forti, Fabio Luis

2013-12-01

170

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?.

2012-06-01

171

Role of PI3K/Akt/mTOR and MEK/ERK pathway in Concanavalin A induced autophagy in HeLa cells.  

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Concanavalin A (Con A), a mannose or glucose specific legume lectin, is well known for its anti-proliferative and cytotoxic effect on different types of cancer cells, through its binding to the membrane receptors leading to a major stimulus for the induction of distinct metabolic responses. Recently it has been also been proved that, Con A induces autophagy in hepatoma cells through internalization and mitochondria mediated pathway involving a mitochondrial interacting protein named Bcl2/E1B-19kDa protein-interacting protein 3 (BNIP3). Through this current endeavor, we propose a membrane associated pathway involved in Con A induced autophagy, taking Human cervical cancer (HeLa) cell as a cancer model. Here, we deciphered the role of membrane mediated phosphatidylinositol 3 kinase (PI3K)/Akt/mTOR (mammalian target of rapamycin) and MEK/Extracellular signal-regulated kinases (ERK) pathway in Con A induced autophagy in HeLa cells. Subsequently, we found that Con A treatment suppresses the PI3K/Akt/mTOR and up regulates the MEK/ERK pathway leading to the activation of autophagy. This study will further help us to understand the mechanism behind the autophagic pathway induced by Con A and simultaneously it will strengthen its effective use as a prospective cancer chemo-therapeutic. PMID:24434245

Roy, Bibhas; Pattanaik, Arup K; Das, Joyjyoti; Bhutia, Sujit K; Behera, Birendra; Singh, Prashant; Maiti, Tapas K

2014-03-01

172

Trichosanthin-induced specific changes of cytoskeleton configuration were associated with the decreased expression level of actin and tubulin genes in apoptotic Hela cells.  

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Trichosanthin (TCS) possesses a broad spectrum of biological and pharmacological activities, including anti-cancer activities through apoptosis pathway. However, little is known about the effects of TCS on the cytoskeleton configuration and expression of actin and tubulin genes in Hela cell apoptosis. In the present study, apoptotic cytoskeleton structures were observed by confocal immunofluorescence microscopy, absolute amounts of actin and tubulin subunit mRNAs were determined by quantitative real-time PCR assays (QRT-PCR). Our results showed that the execution phase of cell apoptosis was a highly coordinated process of cellular reorganization, depolymerized microfilaments (MFs) accumulated in the coarsened cytoplasm and apoptotic bodies, followed by the formation of a ring microtubule (MT) structure beneath the plasma membrane. Importantly, apoptosis occurred by a suppression of actin and tubulin subunit gene expression. In particular, a rapid decrease in the amounts of gamma-actin mRNA preceded that of beta-actin; alpha- and beta-tubulin mRNAs were subsequently down-regulated in the later stage of Hela cell apoptosis. These results suggested that the execution of Hela cell apoptosis induced by TCS accompanied the specific changes of cytoskeleton configuration and, significantly, decreased the expression level of actin and tubulin subunit genes in different stages. PMID:17881009

Wang, Ping; Li, Ji-Cheng

2007-09-15

173

Activation of poly(ADP-ribose) polymerase by sulfur mustard in HeLa cell cultures  

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Poly(ADP-ribose) polymerase (PADPRP) E.C.2.4.2.30 has been proposed to play a key role in the NAD+ depletion following alkylation of DNA in sulfur mustard (HD) exposures. Papirmeister et al. (Fundam Appl Toxicol 5:Sl34, 1985) hypothesized that activation of PADPRP was central to the subsequent depletion of NAD+ and activation of proteolytic enzymes leading to vesication. NAD+ depletion following HD exposure has been previously documented and the results have been used to infer the effect of HD exposure on PADPRP. The present study was undertaken to demonstrate the direct effect of HD on PADPRP activity. HeLa cells culture were used as the model system. At 10 microns HD PADPRP activity was increased above the levels of controls in the first hour. The activity peaked at 4 hrs and by 6 hrs had returned to control levels. The 24-hour level of PADPRP activity was again elevated above the controls. The 100 microns HD exposures had maximal enzymatic response in HeLa cells within the first hour. The level had decreased 40% from the maximum by the second hour reaching a plateau at 30% of the maximum response after 4 hrs. Cells exposed to 100 microns HD showed enzyme levels at or below those seen with the 10 microns dose after 24 hours. The doses of HD used did not decrease viability as measured by trypan blue dye exclusion within 24 hr.

Clark, O.E.; Smith, W.J.

1993-05-13

174

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.

Pincu Rotinberg

2010-01-01

175

Overexpression of cyclin A in human HeLa cells induces detachment of kinetochores and spindle pole/centrosome overproduction.  

Science.gov (United States)

The combination of hydroxyurea (HU) and caffeine has been used for inducing kinetochore dissociation from mitotic chromosomes and for causing centrosome/spindle pole amplification. However, these effects on microtubule organizing centers (MTOCs) are limited to certain cell types. It was reasoned that if the biochemical differences in MTOC behavior between cells following HU treatment could be identified, then critical information concerning the regulation of these organelles would be obtained. During these studies, it was determined that cells from hamster, rat, and deer could be induced to enter mitosis with dissociated kinetochores and to synthesize centrosomes during arrest with HU, while cells from human and mouse could not. Comparisons between human HeLa cells and CHO cells determined that cyclin A levels were depressed in HeLa cells relative to CHO cells following HU addition. Overexpression of cyclin A in HeLa cells converted them to a cell type capable of detaching kinetochores from mitotic chromosomes. Ultrastructural analyses determined that the detached human kinetochores exhibited a normal plate-like morphology and appeared capable of associating with microtubules. In addition, HeLa cells overexpressing cyclin A also overproduced spindle poles during HU arrest, demonstrating that cyclin A activity also is important for centrosome replication during interphase. In summary, elevated cyclin A levels are important for the capacity of cells to be driven into mitosis by caffeine addition, for the ability of cells to progress to mitosis with detached kinetochores, and for centrosome/spindle pole replication. PMID:11734996

Balczon, R C

2001-11-01

176

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

1989-08-01

177

Analysis of histone gene expression during the cell cycle in HeLa cells by using cloned human histone genes.  

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Although it is generally agreed that histone protein synthesis is restricted to the S phase of the cell cycle--and therefore parallels DNA replication--both transcriptional and posttranscriptional levels of control have been invoked. Using blot hybridization with several cloned genomic human histone sequences representing different histone gene clusters as probes, we have assessed the steady-state level of histone RNAs in the nucleus and cytoplasm of G1 and S phase HeLa S3 cells. The represen...

Rickles, R.; Marashi, F.; Sierra, F.; Clark, S.; Wells, J.; Stein, J.; Stein, G.

1982-01-01

178

DNA polymerase ? and ? activity in ?-irradiated HeLa S3 cells  

International Nuclear Information System (INIS)

The acute effects (less than 2 hours) of ?-irradiation on DNA polymerase ? and ? activity in HeLa S3 cells were studied. The enzyme activities were measured in sonicates of the irradiated cells, using an exogenous DNA as template. Both DNA ?- and ?-polymerase activities decreased following irradiation of the cells. Doses as low as 100 rad significantly reduced the activities of the enzymes. While the activities of both DNA polymerases decreased as the dose received by the cells increased, the major reduction in enzyme activity occurred with doses of 100-200 rad. The reduction in DNA ?- and ?-polymerase activities was maximal by 30 min post-irradiation and recovered to control values by 2 hours post-irradiation. (author)

1981-01-01

179

Hyperglycaemia and oxidative stress upregulate HSP60 & HSP70 expression in HeLa cells.  

Science.gov (United States)

Heat Shock Proteins 60 & 70 (HSP60 & HSP70) are intracellular protein that has been shown to be present at elevated levels in systemic circulation in Type 2 Diabetes mellitus (T2DM) patients. Conditions that lead to its secretion, and the mechanism of its translocation from cells, have not yet been defined. The aim of this study was to determine if specific cell stressors associated with T2DM, namely hyperglycaemia and oxidative stress, result in the upregulation of HSP60 in human cells in vitro. Human HeLa cells were grown in media supplemented with 100 mM glucose, 200 ?M hydrogen peroxide (H2O2), and 50 ?M sodium azide. Initially, the effect of these treatments on cell growth rate was examined, with each treatment significantly inhibiting growth rate. LDH and MTT assays were also used to successfully demonstrate that these treatments do not significantly increase cell lysis, but do significantly impair mitochondrial dehydrogenase activity. To confirm this mitochondria specific form of inhibition, DCFDA assay were used to investigate any increases in intracellular reactive oxygen species (ROS) generation. All three treatments resulted in significantly increased ROS generation, with greater ROS production occurring with a greater exposure time. Interestingly, when the protein levels of HSP60 and HSP70 were measured after 3 and 7 days of exposure of the HeLa cells to 100 mM glucose, 200 ?M H2O2, and 50 ?M sodium azide significant induction of these two molecular stress proteins were observed ranging from 2.43-5.08 fold compared to untreated control cells. PMID:24058891

Hall, Luke; Martinus, Ryan D

2013-01-01

180

Monoclonal antibody that inhibits infection of HeLa and rhabdomyosarcoma cells by selected enteroviruses through receptor blockade  

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BALB/c mice were immunized with HeLa cells, and their spleen cells were fused with myeloma cells to produce hybridomas. Initial screening of culture fluids from 800 fusion products in a cell protection assay against coxsackievirus B3 (CB3) and the CB3-RD virus variant yielded five presumptive monoclonal antibodies with three specificities: (i) protection against CB3 on HeLa, (ii) protection against CB3-RD on rhabdomyosarcoma (RD) cells, and (iii) protection against both viruses on the respective cells. Only one of the monoclonal antibodies (with dual specificity) survived two subclonings and was studied in detail. The antibody was determined to have an immunoglobulin G2a isotype and protected cells by blockade of cellular receptors, since attachment of (/sup 35/S)methionine-labeled CB3 was inhibited by greater than 90%. The monoclonal antibody protected HeLa cells against infection by CB1, CB3, CB5, echovirus 6, and coxsackievirus A21 and RD cells against CB1-RD, CB3-RD, and CB5-Rd virus variants. The monoclonal antibody did not protect either cell type against 16 other immunotypes of picornaviruses. The monoclonal antibody produced only positive fluorescence on those cells which were protected against infection, and /sup 125/I-labeled antibody confirmed the specific binding to HeLa and RD cells. The results suggest that this monoclonal antibody possesses some of the receptor specificity of the group B coxsackieviruses.

Crowell, R.L.; Field, A.K.; Schleif, W.A.; Long, W.L.; Colonno, R.J.; Mapoles, J.E.; Emini, E. A.

1986-02-01

 
 
 
 
181

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

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Full Text Available SciELO Venezuela | Language: English 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 lipopolisacá [...] rido de dos serovares de Chlamydia trachomatis (LGV1-LGV2) y usando una coloración Supravital fluorescente (Hoechst 33258) fue posible investigar la respuesta de las células 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 lipopolysaccha [...] ride from two serovars of Chlamydia trachomatis (LGV1-LGV2) and using a nuclear supravital fluorescent stain (Hoechst 33258), it was possible to investigate the apoptotic effect 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.

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

182

Theaflavins depolymerize microtubule network through tubulin binding and cause apoptosis of cervical carcinoma HeLa cells.  

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Here we studied the antiproliferative activity of theaflavins in cervical carcinoma HeLa cells by investigating their effects on cellular microtubules and purified goat brain tubulin. Theaflavins inhibited proliferation of HeLa cells with IC(50) value of 110 ± 2.1 ?g/mL (p = theaflavins act as microtubule depolymerizers. Theaflavins disrupted the microtubule network accompanied by alteration of cellular morphology and also decreased the polymeric tubulin mass of the cells. The polymerization of cold treated depolymerized microtubules in HeLa cells was prevented in the presence of theaflavins. In vitro polymerization of purified tubulin into microtubules was also inhibited by theaflavins with an IC(50) value of 78 ± 2.43 ?g/mL (P theaflavins. PMID:21323312

Chakrabarty, Subhendu; Das, Amlan; Bhattacharya, Abhijit; Chakrabarti, Gopal

2011-03-01

183

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

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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

184

The role of glutathione in the radiosensitive effect induced by treating HeLa cells with sanazole  

International Nuclear Information System (INIS)

Objective: To investigate radiosensitive effect of sanazole on HeLa cells and its relationship with glutathione (GSH). Methods: The anoxia model was made by inflow of nitrogen gas. The survival rate of HeLa cells was observed with method of colony formation after treatment with sanazole and 60Co ? irradiation. Radiosensitive effect was evaluated through measurement of sensitizing enhancement ratio (SER) resulted from single-target multi-hit model. The GSH content in these HeLa cells was determined by the tetra-oxypyrimidine UV-spectrophotometer method to explore the mechanism of radiosensitive effect. Results: SER was more than 1.4. The concentration of GSH decreased significantly with increasing concentration of sensitizer and dose of radiation, especially under anoxia condition. Conclusions: Sanazole has significant radiosensitive effect and decrease in GSH content resulted from combination with 60Co ? irradiation may be one of its radiosensitive mechanisms

2001-12-01

185

The effect of aclacinon on macromolecular syntheses in HeLa cells  

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The effects of aclacinon (ACR, aclacinomycin A) on DNA and RNA syntheses in HeLa cells were examined by measuring incorporation of labeled precursors. The cells were treated with ACR at various concentrations for 2 or 24 hr, followed by the incubation with "3H-TdR (1 ?Ci/ml) or "3H-UR (2.5 ?Ci/ml) for 1 hr. The uptakes of "3H-TdR and "3H-UR immediately after the treatment with ACR for 2 hr were inhibited in proportion to the concentration of ACR. The drug proved to be a more potent inhibitor to the RNA synthesis than to the DNA synthesis. Fifty percent inhibitory concentration values (IC_5_0) of ACR for "3H-TdR and "3H-UR uptake in HeLa RC-355 cells were 0.39 ?g/ml and 0.16 ?g/ml, respectively. The ratio of IC_5_0 was calculated as about 2.4. The uptake of "3H-TdR and "3H-UR were significantly inhibited at lower concentrations when treated with ACR for 24 hr than when treated for 2 hr. In the experiments on the recovery of DNA and RNA syntheses, the uptakes of "3H-TdR were inhibited to about 70 % and 40 % of the control, respectively, by the ACR treatment at 0.2 ?g/ml and 0.5 ?g/ml for 2 hr, and the recovery of uptake was observed about 4 hr after the removal of ACR from the cultures. On the other hand, in cases of 24 hr exposure to ACR, DNA and RNA syntheses have remained low even 24 hr after the removal of ACR. After a log-phase culture of HeLa RC-355 cells was treated with ACR for 24 hr, the restraint of the growth by ACR was observed at the lowest concentration tested (0.05 ?g/ml) and almost completely at 0.2 ?g/ml of ACR. (Namekawa, K.)

1985-01-01

186

Autoradiography of DNA from Hela cells under normal conditions and after treatment with hydroxyurea  

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The results are presented of the first stage of the elaboration of the novel autoradiographic technique for studying the replication of DNA fibers from nonsynchronized Hela cell cultures under normal conditions and after treatment with hydroxyurea. The preparations were covered with liquid nuclear emulsion Ilford L_4. Exposure was carried out for 3 months at 4 deg C. After development, the autoradiograms were recorded quantitatively, and the length of the individual replicative segments was measured by means of an object micrometers. For each group (control and experimental) 100 segments from different cells were recorded. The results obtained were subjected to mathematical-statistical processing for determining the standard deviation. The application of hidroxyurea highly reduces the replicative elements, i.e. it actually inhibits DNA synthesis. This inhibition is due to reduction in the production of the four endogenous deoxynucleotides and affects the length of growth of the DNA chain, but the interreplicative distance as well

1984-01-01

187

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

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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

2011-10-01

188

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

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Full Text Available SciELO Brazil | Language: English 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 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.

189

STAT3 is involved in phosphatidic acid-induced Bcl-2 expression in HeLa cells  

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Phosphatidic acid (PA), the product of a PLD-mediated reaction, is a lipid second messenger that participates in various intracellular signaling events and is known to regulate a growing list of signaling proteins. We found that Bcl-2 was upregulated by PA treatment in HeLa cells. However, how PA upregulates Bcl-2 expression has not yet been studied. In this study, we tried to discover the mechanisms of Bcl-2 up-regulation by PA treatment in HeLa cells. Treatment with PA resulted in significa...

Choi, Hye-jin; Lee, Jung Han; Park, Shin-young; Cho, Ju Hwan; Han, Joong-soo

2009-01-01

190

Radiation-induced thymine base damage and its excision repair in active and inactive chromatin of HeLa cells  

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The extent of production and excision repair of 5,6-dihydroxydihydrothymine type base (t') damage was determined in transcriptionally active and inactive chromatin of HeLa cells after exposure to 6.8 MeV electrons. It was observed that not only the yield but also rate of repair of t' products was greater in the active chromatin compared to the inactive chromatin of HeLa cells. The results strongly indicate that the conformation of chromatin is an important factor in determining the sensitivity to radiation damage and accessibility to enzymes required for repair of such damage. (author)

1985-11-01

191

Effects of triterpenes from Ganoderma lucidum on protein expression profile of HeLa cells.  

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To elucidate the cytotoxicity mechanism of Ganoderma triterpenes, a chemoproteomic study using five purified ganoderic acids, ganoderic acid F (GAF), ganoderic acid K (GAK), ganoderic B (GAB), ganoderic acid D (GAD) and ganoderic acid AM1 (GAAM1) was conducted. GAF, GAK, GAB, GAD and GAAM1 treatment for 48 h inhibited the proliferation of HeLa human cervical carcinoma cells with IC(50) values of 19.5+/-0.6 microM, 15.1+/-0.5 microM, 20.3+/-0.4 microM, 17.3+/-0.3 microM, 19.8+/-0.7 microM, respectively. The protein expression profiles of HeLa cells treated with each ganoderic acid at dose of 15 microM for 48 h were checked using two-dimensional electrophoresis (2-DE). The possible target-related proteins of ganoderic acids, i.e. proteins with same change tendency in all five ganoderic acids-treated groups compared with control, were identified using MALDI-TOF MS/MS. Twelve proteins including human interleukin-17E, eukaryotic translation initiation factor 5A (eIF5A), peroxiredoxin 2, ubiquilin 2, Cu/Zn-superoxide dismutase, 14-3-3 beta/alpha, TPM4-ALK fusion oncoprotein type 2, PP2A subunit A PR65-alpha isoform, nucleobindin-1, heterogeneous nuclear ribonucleoprotein K, reticulocalbin 1 and chain A of DJ-1 protein were identified. Ganoderic acids might exert their cytotoxicity by altering proteins involved in cell proliferation and/or cell death, carcinogenosis, oxidative stress, calcium signaling and ER stress. PMID:20092987

Yue, Q-X; Song, X-Y; Ma, C; Feng, L-X; Guan, S-H; Wu, W-Y; Yang, M; Jiang, B-H; Liu, X; Cui, Y-J; Guo, D-A

2010-07-01

192

Povidone-iodine-induced cell death in cultured human epithelial HeLa cells and rat oral mucosal tissue.  

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Abstract Although povidone-iodine (PVP-I) has been used as a gargle since 1956, its effectiveness and material safety have been remained controversial. The aim of this study was to investigate the toxicity of PVP-I to epithelial cells in a concentration range significantly lower than that used clinically. Study design was in vitro laboratory investigations and in vivo histological and immunologic analysis. We examined the effects of PVP-I at concentrations of 1?×?10(-2) to 1?×?10(3?)?M and 1?×?10(-4) to 1?×?10??M on HeLa cells as a model of epithelial cells and rat oral mucosa, respectively, after 1 or 2 days of exposure. Annexin V/FLUOS was used to distinguish live, apoptotic and necrotic cells. The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) method was also used to observe whether apoptotic epithelial cells exist in rat oral mucosa after 1 day of exposure of PVP-I. HeLa cells developed concentration-dependent cytotoxicity, and epithelium of rat oral mucosa was thinned in a concentration-dependent manner. HeLa cell apoptosis increased after 1?×?10(0)??M of PVP-I exposure for 2 days. In the TUNEL method, many apoptotic epithelial cells were observed in the rat oral mucosa after 1 day of exposure to diluted 1?×?10(-2)??M of PVP-I, but minimal apoptotic epithelial cells were observed using 1?×?10(-3)??M of PVP-I. Our findings suggest that exposure to PVP-I, of which concentrations are even lower than those used clinically, causes toxicity in epithelial cells. This knowledge would help us better understand the risk of the use of PVP-I against mucosa. PMID:24219135

Sato, So; Miyake, Masao; Hazama, Akihiro; Omori, Koichi

2014-07-01

193

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.

Jan Oleszczuk

2010-05-01

194

Actin deficiency induces cofilin phosphorylation: proteome analysis of HeLa cells after beta-actin gene silencing.  

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Actin-binding proteins regulate the dynamic structure and function of actin filaments in the cell. Much is known about how manipulation of the actin-binding proteins affects the structure and function of actin filaments; however, little is known about how manipulation of actin in the cell affects actin-binding proteins. We addressed this question by utilizing two technologies: RNA interference and 2-dimensional gel electrophoresis. We knocked down beta-actin expression in HeLa cells using short interfering RNA and applied 2-DGE to examine alterations in the HeLa cell proteome. We revealed a 2-5 fold increases of four protein spots on 2-D gels and identified these proteins by mass spectrometry. Three of the four proteins were actin-binding proteins, including cofilin, which promotes both disassembly and assembly of actin filaments but becomes inactivated when phosphorylated. Further examination revealed that the cofilin total protein level barely increased, but the phosphorylated cofilin level increased dramatically in HeLa cells after beta-actin siRNA treatment. These results suggest that in response to siRNA-induced beta-actin deficiency HeLa cells inactivate cofilin by phosphorylation rather than down-regulate its protein expression level. This study also demonstrates that the combination of RNA interference and 2-dimensional gel electrophoresis technologies provides a valuable method to study protein interactions in a specific cellular pathway. PMID:17123313

Liu, Ning; Academia, Katrina; Rubio, Teresa; Wehr, Tim; Yeck, Todd; Jordan, Liz; Hamby, Keith; Paulus, Aran

2007-02-01

195

Nutritional and Metabolic Requirements for the Infection of HeLa Cells by Salmonella enterica Serovar Typhimurium  

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Salmonella is the causative agent of a spectrum of human and animal diseases ranging from gastroenteritis to typhoid fever. It is a food - and water - borne pathogen and infects via ingestion followed by invasion of intestinal epithelial cells and phagocytic cells. In this study we employed a mutational approach to define the nutrients and metabolic pathways required by Salmonella enterica serovar Typhimurium during infection of a human epithelial cell line (HeLa). We deleted the key glycolytic genes, pfkA and pfkB to show that S. Typhimurium utilizes glycolysis for replication within HeLa cells; however, glycolysis was not absolutely essential for intracellular replication. Using S. Typhimurium strains deleted for genes encoding components of the phosphotransferase system and glucose transport, we show that glucose is a major substrate required for the intracellular replication of S. Typhimurium in HeLa cells. We also deleted genes encoding enzymes involved in the utilization of gluconeogenic substrates and the glyoxylate shunt and show that neither of these pathways were required for intracellular replication of S. Typhimurium within HeLa cells.

Rice, Christopher J.; Ramachandran, Vinoy K.; Kelly, David J.; Thompson, Arthur

2014-01-01

196

Caveolae-mediated endocytosis of biocompatible gold nanoparticles in living Hela cells  

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Efficient intracellular delivery of gold nanoparticles (AuNPs) and unraveling the mechanism underlying the intracellular delivery are essential for advancing the applications of AuNPs toward in vivo imaging and therapeutic interventions. We employed fluorescence microscopy to investigate the internalization mechanism of small-size AuNPs by living Hela cells. Herein, we found that the caveolae-mediated endocytosis was the dominant pathway for the intracellular delivery of small-size AuNPs. The intracellular delivery was suppressed when we depleted the cholesterol with methyl-β-cyclodextrin (M beta CD); in contrast, the sucrose that disrupts the formation of clathrin-mediated endocytosis did not block the endocytosis of AuNPs. Meanwhile, we examined the intracellular localization of AuNPs in endocytic vesicles by fluorescent colocalization. This work would provide a potential technique to study the intracellular delivery of small-size nanoparticles for biomedical applications.

Hao, Xian

2012-01-01

197

Cytotoxic activity of proteins isolated from extracts of Corydalis cava tubers in human cervical carcinoma HeLa cells  

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Background Corydalis cava Schweigg. & Koerte, the plant of numerous pharmacological activities, together with the studied earlier by our group Chelidonium majus L. (Greater Celandine), belong to the family Papaveraceae. The plant grows in Central and South Europe and produces the sizeable subterraneous tubers, empty inside, which are extremely resistant to various pathogen attacks. The Corydalis sp. tubers are a rich source of many biologically active substances, with the extensive use in European and Asian folk medicine. They have analgetic, sedating, narcotic, anti-inflammatory, anti-allergic and anti-tumour activities. On the other hand, there is no information about possible biological activities of proteins contained in Corydalis cava tubers. Methods Nucleolytic proteins were isolated from the tubers of C. cava by separation on a heparin column and tested for DNase activity. Protein fractions showing nucleolytic activity were tested for cytotoxic activity in human cervical carcinoma HeLa cells. Cultures of HeLa cells were conducted in the presence of three protein concentrations: 42, 83 and 167 ng/ml during 48 h. Viability of cell cultures was appraised using XTT colorimetric test. Protein fractions were separated and protein bands were excised and sent for identification by mass spectrometry (LC-ESI-MS/MS). Results The studied protein fractions showed an inhibiting effect on mitochondrial activity of HeLa cells, depending on the administered dose of proteins. The most pronounced effect was obtained with the highest concentration of the protein (167 ng/ml) - 43.45 ± 3% mitochondrial activity of HeLa cells were inhibited. Mass spectrometry results for the proteins of applied fractions showed that they contained plant defense- and pathogenesis-related (PR) proteins. Conclusions The cytotoxic effect of studied proteins toward HeLa cell line cells has been evident and dependent on increasing dose of the protein. The present study, most probably, represents the first investigations on the effect of purified PR proteins from tuber extracts of a pharmacologically active plant on cell lines.

2010-01-01

198

Structural and physicochemical aspects of silica encapsulated ZnO quantum dots with high quantum yield and their natural uptake in HeLa cells.  

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Photoluminescent semiconductor quantum dots (QDs) are of significant interest for bioimaging and fluorescence labeling. In this regard, we describe here the design of high sensitivity and high specificity non-toxic ZnO QDs (?5 nm) with long-term stability of up to 12 months. The embedding of ZnO QDs on silica nanospheres led to significant increase in photoluminescence intensity rendering them highly bright QD-based probes. The QDs were characterized in vitro with respect to cancer cells (HeLa) and evaluated in terms of viability, fluorescence and cytoskeletal organization. The immobilization of ZnO QDs on silica nanospheres promoted the internalization and enhanced fluorescence emission of HeLa cells. The fluorescence emission from QDs was stable for 3 days, indicating excellent stability toward photobleaching. Cytoskeletal reorganization was observed after internalization of QDs such that the ZnO QDS on silica nanospheres resulted in broadening of the actin cytoskeleton. The study underscores that ZnO QDs immobilized on Si nanospheres are promising for tracking cancer cells in cell therapy. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 102A: 2934-2941, 2014. PMID:24115677

Depan, D; Misra, R D K

2014-09-01

199

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

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

2014-01-01

200

Effect of Lon protease knockdown on mitochondrial function in HeLa cells.  

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ATP-dependent proteases are currently emerging as key regulators of mitochondrial functions. Among these proteolytic systems, Lon protease is involved in the control of selective protein turnover in the mitochondrial matrix. In the absence of Lon, yeast cells have been shown to accumulate electron-dense inclusion bodies in the matrix space, to loose integrity of mitochondrial genome and to be respiratory deficient. In order to address the role of Lon in mitochondrial functionality in human cells, we have set up a HeLa cell line stably transfected with a vector expressing a shRNA under the control of a promoter which is inducible with doxycycline. We have demonstrated that reduction of Lon protease results in a mild phenotype in this cell line in contrast with what have been observed in other cell types such as WI-38 fibroblasts. Nevertheless, deficiency in Lon protease led to an increase in ROS production and to an accumulation of carbonylated protein in the mitochondria. Our study suggests that Lon protease has a wide variety of targets and is likely to play different roles depending of the cell type. PMID:24355201

Bayot, Aurélien; Gareil, Monique; Chavatte, Laurent; Hamon, Marie-Paule; L'Hermitte-Stead, Caroline; Beaumatin, Florian; Priault, Muriel; Rustin, Pierre; Lombès, Anne; Friguet, Bertrand; Bulteau, Anne-Laure

2014-05-01

 
 
 
 
201

Long-term effects of methylmercury (II) on the viability of HeLa S3 cells  

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The communication reports on experiments in which asynchronous HeLa S3 suspension-culture cells were exposed to methylmercury for periods lasting up to 432 hr or almost 21 generations. The cell response is expressed in terms of cell viability as given by the trypan blue dye exclusion test. Also presented are some preliminary data on the extent and rate of toxicant uptake by the cells using /sup 203/Hg-labeled methylmercury as a marker.

Gruenwedel, D.W.; Friend, D.

1980-09-01

202

Comparison between the comet assay and fast micromethod (R) for measuring DNA damage in HeLa cells  

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The sensitivity and precision of the single cell gel electrophoresis (Comet) assay and Fast Micromethod(R) for DNA damage determinations in human HeLa cell line were compared. The first assay allows analysis of DNA breaks in individual cells while the second is a rapid and convenient procedure for DNA breaks determination in cell suspensions on single microplates. Both assays detect DNA strand breaks, alkali-labile sites and transient breaks occurring at sites of ongoing repair and might be a...

2002-01-01

203

Cytotoxicity of different extracts of arial parts of Ziziphus spina-christi on Hela and MDA-MB-468 tumor cells  

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Background: It has been shown that plants from the family Rhamnaceae possess anticancer activity. In this study, we sought to determine if Ziziphus spina-christi, a species from this family, has cytotoxic effect on cancer cell lines. Materials and Methods: Using maceration method, different extracts of leaves of Z. spina-christi were prepared. Hexane, chloroform, chloroform-methanol (9:1), methanol-water (7:1) methanol, butanol and water were used for extraction, after preliminary phytochemical analyses were done. The cytotoxic activity of the extracts against Hela and MDA-MB-468 tumor cells was evaluated by MTT assay. Briefly, cells were seeded in microplates and different concentrations of extracts were added. After incubation of cells for 72 h, their viability was evaluated by addition of tetrazolium salt solution. After 3 h medium was aspirated, dimethyl sulfoxide was added and absorbance was determined at 540 nm with an ELISA plate reader. Extracts were considered cytotoxic when more than 50% reduction on cell survival was observed. Results: Hexane, chloroform, chloroform-methanol, butanol, methanol-water and aqueous extracts of Z. spina-christi significantly and concentration-dependently reduced viability of Hela and MAD-MB-468 cells. In the both cell lines, chloroform-methanol extract of Z. spina-christi was more potent than the other extracts. Results: From the finding of this study it can be concluded that Z. spina-christi is a good candidate for further study for new cytotoxic agents.

Jafarian, Abbas; Zolfaghari, Behzad; Shirani, Kobra

2014-01-01

204

Cytotoxic effect against HeLa cells of polysaccharides from the lichen Ramalina celastri.  

Science.gov (United States)

The most active polysaccharides which show anti-tumoral activity are (1-->3)-beta-D-glucans, branched or not at O-6. Since these structures are sometimes poorly soluble in aqueous media, alpha-D-glucans and their chemical derivatives, which are more soluble, were also studied. The present object is to observe morphological alterations in HeLa cells caused by two different polysaccharides obtained from the lichen Ramalina celastri, which are (1-->3),(1-->4)-linked alpha-D-glucan and its sulphated derivative. The cells were incubated in Eagle's medium in the absence or presence of each polysaccharide and routinely processed and analysed by light and electron microscopy. Even though the alpha-D-glucan altered the cellular volume, cytoplasmic densities, and mitosis, the resulting monolayer was similar to the control. TEM analysis showed cytoplasmic blebbing and the presence of an amorphous electron-dense material free in the cytoplasm and interior membranes. The enhanced injury caused by the sulphated derivative was apparent, altering cell adhesion and causing cell aggregation. Nuclear modifications such as fragmentation and condensation of chromatin under the nuclear envelope, which showed to be convoluted, suggested the occurrence of cell death by apoptosis. PMID:9397587

Leão, A M; Buchi, D F; Iacomini, M; Gorin, P A; Oliveira, M B

1997-10-01

205

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

206

Interaction between Yersinia pseudotuberculosis and the HeLa cell surface.  

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The adherence of Yersinia pseudotuberculosis to the surface of HeLa cells at 4 degrees C was studied. This temperature allows adhesion of bacteria but prevents engulfment. Adhesion between the bacteria and the cells was not dependent upon the presence of serum, Ca2+ or Mg2+ in the medium. Maximum adhesion was obtained at pH 6.5-7.9 and pretreatment of the cells with formaldehyde or glutaraldehyde inhibited the attachment of the bacteria. The interaction between the bacteria and the cell surface seems to involve cellular processes that are mostly microvilli. An intimate association between the bacteria and the cellular glycocalyx was found. Three virulent bacterial strains adhered more easily to the cell surface than five avirulent strains. Maximum adherence was obtained with bacteria from late logarithmic and early stationary phases of growth. The bacteria gradually lose their adhesive property when cultivated for several generations at 37 degrees C in nutrient broth but not when cultivated at 20 degrees C. Treatment of the bacteria with protease IV from Streptomyces caespitosus markedly reduced the efficiency of attachment. PMID:6876136

Brunius, G; Bölin, I

1983-08-01

207

Effects of BSO and DEM on thiol-level and radiosensitivity in HeLa cells  

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Reduction of the intracellular GSH and NPSH levels in HeLa cells by BSO and DEM treatments was determined. As parameters for radiation damage, single and double strand DNA breaks (ssb and dsb) and cell killing were used. Furthermore, repair of ssb and dsb were followed in the first 30 and 120 min after radiation, respectively. BSO and DEM treatment gave a small sensitization for the 3 types of radiation damage (ssb, dsb and cell killing) in aerobic condition. In hypoxic condition the sensitizing effect of both compounds on dsb was larger than the effect on ssb. Pretreatment with BSO and DEM had no influence on repair of ssb and dsb when cells were irradiated in air, but when cells were irradiated in hypoxia, repair was somewhat inhibited after pretreatment with DEM. It can be postulated that a reduction of the intracellular GSH level by BSO and DEM treatment affects cellular radiosensitivity both by a competitive mechanism between GSH and O2 and by inhibition of enzymatic repair of DNA breaks, the latter only in the case of DEM treatment

1984-01-01

208

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

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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

2013-01-01

209

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

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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. Biggiogera

2011-04-01

210

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)

1968-08-01

211

Radiation-induced association of beta-glucuronidase with purified nuclei from irradiated MOLT-4 and HeLa cells  

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Beta-glucuronidase, a lysosomal marker enzyme, associates with purified nuclei from HeLa and MOLT-4 cell lines in a radiation dose-dependent manner, up to 300 cGy in MOLT-4 cells, and 1000 cGy in HeLa cells. In MOLT-4 cells (200-cGy exposure), there is a significant increase in beta-glucuronidase activity detected in the nuclear fraction 24 h postirradiation with a maximum association occurring at 72 h. In HeLa cells (1000-cGy exposure), a significant association is first detected 24 h postirradiation with a maximum association at 48 h. The association is not the result of nonspecific contamination occurring during nuclei purification since nuclei from irradiated cells show no greater levels of plasma membrane marker and mitochondrial marker than controls. The nature of the association remains unclear, but activity is not removed by detergents used in the nuclei isolation procedure, and incubation of the nuclei with EDTA reverses the association only modestly. Exposure of nuclei from irradiated cells to anisotonic buffers also results in only a small decrease in beta-glucuronidase activity associated with the nuclei. These observations suggest that lysosomal hydrolases become intimately associated with the nuclei of irradiated cells

1989-01-01

212

Radiosensitivity of polyamine-depleted HeLa cells and modulation by the aminothiol WR-1065  

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The radiosensitivity of cultured HeLa cells was increased upon depletion of the natural cellular polyamines putrescine, spermidine and spermine through treatment of cultures with inhibitors of polyamine biosynthesis. This increased radiosensitivity was manifested as a decrease in the D_0 and by the absence of a shoulder in the survival curves. However, our previous studies have shown that the initial yield of X-ray-induced DNA damage did not appear to be elevated in polyamine-depleted cells. In addition, polyamine-depleted cells exhibited markedly altered X-ray-induced changes in the distribution of cells in the phases of the cell cycle characterized by increased time of onset and lengthened duration of G_2-phase delay. Addition of polyamines to cultures for short periods prior to irradiation restored normal radioresistence and reversed the anomalous features of the G_2-phase delay profile. Polyamine supplementation experiments as well as studies in which combinations of inhibitors were employed to modulate specific polyamine levels suggest that spermidine may play a primary role in governing cellular radioresponsiveness. The radioprotective aminothiol WR-1065 protected normal and polyamine-depleted cells to a proportionately similar extent (protection factor of 2.4 and 2.8, respectively) but had no apparent ability to restore the shoulder or alter the G_2-phase delay markedly in polyamine-depleted cells. The findings reported here extend our previous observations that polyamine depletion results in a compromised ability to respond to X irradiation and suggest that a defect in repair and/or the G_2-phase delay response may be the determining factors. 34 refs., 8 figs., 3 tabs

1994-01-01

213

Radiosensitivity of polyamine-depleted HeLa cells and modulation by the aminothiol WR-1065  

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The radiosensitivity of cultured HeLa cells was increased upon depletion of the natural cellular polyamines putrescine, spermidine and spermine through treatment of cultures with inhibitors of polyamine biosynthesis. This increased radiosensitivity was manifested as a decrease in the D{sub 0} and by the absence of a shoulder in the survival curves. However, our previous studies have shown that the initial yield of X-ray-induced DNA damage did not appear to be elevated in polyamine-depleted cells. In addition, polyamine-depleted cells exhibited markedly altered X-ray-induced changes in the distribution of cells in the phases of the cell cycle characterized by increased time of onset and lengthened duration of G{sub 2}-phase delay. Addition of polyamines to cultures for short periods prior to irradiation restored normal radioresistence and reversed the anomalous features of the G{sub 2}-phase delay profile. Polyamine supplementation experiments as well as studies in which combinations of inhibitors were employed to modulate specific polyamine levels suggest that spermidine may play a primary role in governing cellular radioresponsiveness. The radioprotective aminothiol WR-1065 protected normal and polyamine-depleted cells to a proportionately similar extent (protection factor of 2.4 and 2.8, respectively) but had no apparent ability to restore the shoulder or alter the G{sub 2}-phase delay markedly in polyamine-depleted cells. The findings reported here extend our previous observations that polyamine depletion results in a compromised ability to respond to X irradiation and suggest that a defect in repair and/or the G{sub 2}-phase delay response may be the determining factors. 34 refs., 8 figs., 3 tabs.

Snyder, R.D.; Schroeder, K.K. [Marion Merrell Dow Research Institute, Cincinnati, OH (United States)

1994-01-01

214

High LET radiation enhances nocodazole induced cell death in HeLa cells through mitotic catastrophe and apoptosis  

International Nuclear Information System (INIS)

To understand how human tumor cells respond to the combined treatment with nocodazole and high linear energy transfer (LET) radiation, alterations in cell cycle, mitotic disturbances and cell death were investigated in the present study. Human cervix carcinoma HeLa cells were exposed to nocodazole for 18 h immediately followed by high LET iron ion irradiation and displayed a sequence of events leading to DNA damages, mitotic aberrations, interphase restitution and endocycle as well as cell death. A prolonged mitotic arrest more than 10 h was observed following nocodazole exposure, no matter the irradiation was present or not. The occurrence of mitotic slippage following the mitotic arrest was only drug-dependent and the irradiation did not accelerate it. The amount of polyploidy cells was increased following mitotic slippage. No detectable G2 or G1 arrest was observed in cells upon the combined treatment and the cells reentered the cell cycle still harboring unrepaired cellular damages. This premature entry caused an increase of multipolar mitotic spindles and amplification of centrosomes, which gave rise to lagging chromosomal material, failure of cytokinesis and polyploidization. These mitotic disturbances and their outcomes confirmed the incidence of mitotic catastrophe and delayed apoptotic features displayed by terminal-transferased UTP- nick end-labeling (TUNEL) method after the combined treatment. These results suggest that the addition of high-LET iron ion irradiation to nocodazole enhanced mitotic catastrophe and delayed apoptosis in HeLa cells. These might be important cell death mechanisms involved in tumor cells in response to the treatment of antimitotic drug combined with high LET radiation. (author)

2011-07-01

215

Depletion of cellular poly (A) binding protein prevents protein synthesis and leads to apoptosis in HeLa cells  

International Nuclear Information System (INIS)

Highlights: ? Depletion of cellular PABP level arrests mRNA translation in HeLa cells. ? PABP knock down leads to apoptotic cell death. ? PABP depletion does not affect transcription. ? PABP depletion does not lead to nuclear accumulation of mRNA. -- Abstract: The cytoplasmic poly (A) binding protein (PABP) is important in mRNA translation and stability. In yeast, depletion of PABP leads to translation arrest. Similarly, the PABP gene in Drosophila is important for proper development. It is however uncertain, whether mammalian PABP is essential for mRNA translation. Here we showed the effect of PABP depletion on mRNA metabolism in HeLa cells by using a small interfering RNA. Our results suggest that depletion of PABP prevents protein synthesis and consequently leads to cell death through apoptosis. Interestingly, no detectable effect of PABP depletion on transcription, transport and stability of mRNA was observed.

2011-05-13

216

Depletion of cellular poly (A) binding protein prevents protein synthesis and leads to apoptosis in HeLa cells  

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Highlights: {yields} Depletion of cellular PABP level arrests mRNA translation in HeLa cells. {yields} PABP knock down leads to apoptotic cell death. {yields} PABP depletion does not affect transcription. {yields} PABP depletion does not lead to nuclear accumulation of mRNA. -- Abstract: The cytoplasmic poly (A) binding protein (PABP) is important in mRNA translation and stability. In yeast, depletion of PABP leads to translation arrest. Similarly, the PABP gene in Drosophila is important for proper development. It is however uncertain, whether mammalian PABP is essential for mRNA translation. Here we showed the effect of PABP depletion on mRNA metabolism in HeLa cells by using a small interfering RNA. Our results suggest that depletion of PABP prevents protein synthesis and consequently leads to cell death through apoptosis. Interestingly, no detectable effect of PABP depletion on transcription, transport and stability of mRNA was observed.

Thangima Zannat, Mst.; Bhattacharjee, Rumpa B. [Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G2W1 (Canada); Bag, Jnanankur, E-mail: jbag@uoguelph.ca [Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G2W1 (Canada)

2011-05-13

217

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

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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

2013-06-01

218

Effect of X-irradiation of clonogenic HeLa cells on the genome mutation frequencies in their progenies  

International Nuclear Information System (INIS)

Irradiation of clonogenic Hela cells with 100-350 R doses results in the increase of general frequency of genome mutations from (20.7+-0.4)x10"-"2 up to (24.8+-0.4)x10"-"2-(31.9+-0.3)x10"-"2 on a cell per a generation. The increase occurs mainly at the expense of hyperploid mutants, whereas frequency of appearance of cells with reduced number of chromosomes (hypoploids) does not change reliably. For Hela culture, used in experiments, a very high heterogeneity of cells on DNA content in interfase nuclei and a very high level of spontaneous frequency of genome mutation are characteristical, that should be taken into account during the analysis of obtained results

1980-01-01

219

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

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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

2003-12-01

220

Isolation and structural characterization of cap-binding proteins from poliovirus-infected HeLa cells.  

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In poliovirus-infected HeLa cells, poliovirus RNA is translated at times when cellular mRNA translation is strongly inhibited. It is thought that this translational control mechanism is mediated by inactivation of a cap-binding protein complex (comprising polypeptides of 24 [24-kilodalton cap-binding protein], 50, and approximately 220 kilodaltons). This complex can restore the translation of capped mRNAs in extracts from poliovirus-infected cells. We have previously shown that the virally in...

Lee, K. A.; Edery, I.; Sonenberg, N.

1985-01-01

 
 
 
 
221

[Effects of colchicine, vinblastine and mercury p-hydroxybenzoate on the multiplication of adenovirus 5 in HeLa cells].  

Science.gov (United States)

After 30 min incubation at 37 with 10(-4) M PHMB (p-hydroxymercuribenzoate), monolayers of HeLa cells are not affected; after 3 h exposure to 10(-5) M colchicin or 2.5 X 10(-5) M vinblastine, HeLa cells are not altered morphologically but the number of cells is decreasing between 1 and 2 days post treatment. When cells are kept in suspension after trypsinization, their ability to adhere to the Petri dishes is not altered with 10(-4) M PHMB but only 50% of the cells are able to attach after treatment with colchicine or vinblastine. Adenovirus type 5 is unsensitive to the effect of colchicine and vinblastine at various concentrations, after 3 h incubation at 37. With 10(-3) M PHMB, the virus is inactivated, infectivity and hemagglutinating activity are almost abolished but the particles are unsensitive to pancreatic DNase, so that the structure of the viral particle is intact. This effect is reduced with 10(-4) M PHMB. The adenovirus 5 multiplication is not affected when HeLa cells are treated with 10(-5) M colchicine or 2.5 X 10(-5) M vinblastine, before and during adsorption of the virus. The viral production decreases after 24 h treatment with colchicine or vinblastine. Treatment with 10(-4) M PHMB does not influence intra- or extracellular viral yield. PMID:1268311

Chardonnet, Y; Lyon, M; Sohier, R

1976-01-01

222

A combined treatment of HeLa cells with the farnesyl protein transferase inhibitor L-744,832 and cisplatin significantly increases the therapeutic effect as compared to cisplatin monotherapy.  

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Activating mutations of Ras that frequently occur during malignant transformation, enhance growth-promoting signal transduction, allowing cells to bypass stringent control of cell cycle progression, thereby rendering them highly proliferative. Abundantly expressed c-Ha-ras protein in human cervical HeLa cells is farnesylated and attached to the plasma membrane, inducing enhanced signal transduction. Exposure of HeLa cells to cisplatin very efficiently inhibits cell proliferation and induces apoptosis. Unfortunately, high doses of cisplatin are strongly cytotoxic, therefore, an alternative therapeutic strategy allowing dose reduction of cisplatin by inhibition of farnesylation could increase the curative effects of cisplatin, thereby benefiting cancer patients. We used two inhibitors of farnesyl protein transferase (FPTase), FTI, and L-744,832, to sensitize HeLa cells to the action of cisplatin. The combined administration of cisplatin and inhibitors of FPTase increased the cytostatic potency of cisplatin. L-744,832 exhibited a stronger synergistic effect in combination with cisplatin than FTI. Moreover, the efficiency of the combined therapy strongly depended on the treatment regimen: The highest efficiency was achieved after combined treatment for 24 h and post-incubation with an inhibitor of FPTase for 48 h. Following this optimized treatment, apoptosis was induced in approximately 50% of HeLa cells treated with 1 microM cisplatin, representing approximately a threefold increase as compared to cisplatin monotherapy. Combined treatment of HeLa cells with cisplatin and inhibitors of FPTase significantly increases the efficacy of the therapy and allows to reduce the dose of cisplatin. Importantly, best therapeutic effects can be achieved by post-treatment with inhibitors of FPTase. PMID:18022825

Wesierska-Gadek, Józefa; Kramer, Matthias P; Schmid, Gerald

2008-05-01

223

TOF-SIMS 3D imaging of native and non-native species within HeLa cells.  

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In this study, a non-native chemical species, bromodeoxyuridine (BrdU), was imaged within single HeLa cells using time-of-flight secondary ion mass spectrometry (TOF-SIMS). z-corrected 3D images were reconstructed that accurately portray the distribution of intracellular BrdU as well as other intracellular structures. The BrdU was localized to the nucleus of cells, whereas structures composed of CxHyOz(-) species were located in bundles on the periphery of cells. The CxHyOz(-) subcellular features had a spatial resolution at or slightly below a micrometer (900 nm), as defined by the distance between the 16% and 84% intensities in a line scan across the edge of the features. Additionally, important parameters influencing the quality of the HeLa cell 3D images were investigated. Atomic force microscopy measurements revealed that the HeLa cells were sputtered at a rate of approximately 4 nm per 10(13) C60(+) ions/cm(2) at 10 keV and a 45° incident angle. Optimal 3D images were acquired using a Bi3(+) liquid metal ion gun operating in the simultaneous high mass and spatial resolution mode. PMID:24131300

Brison, Jeremy; Robinson, Michael A; Benoit, Danielle S W; Muramoto, Shin; Stayton, Patrick S; Castner, David G

2013-11-19

224

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

225

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

226

Purification and characterization of the glycoprotein hormone ?-subunit-like material secreted by HeLa cells  

International Nuclear Information System (INIS)

The protein secreted by HeLa cells that cross-reacts with antiserum developed against the ?-subunit of human chorionic gonadotropin (hCG) has been purified approximately 30,000-fold from concentrated culture medium by organic solvent fractionation followed by ion exchange, gel filtration, and lectin affinity chromatography. The final preparation had a specific activity (by RIA) of 6.8 x 105 ng of ?/mg of protein and appeared homogeneous by electrophoresis on reducing/denaturing polyacrylamide gels (SDS-PAGE). Amino acid analysis indicated that HeLa-? had a composition very similar to that of the urinary hCG ?-subunit. However, comparison of hCG-? and HeLa-? demonstrated that the tumor-associated subunit was not identical with its normal counterpart. The purified tumor protein had an apparent molecular weight greater than that of the urinary ?-subunit when analyzed by SDS-PAGE, and this difference was even greater when a partially purified preparation was examined by an immunoblot technique (Western). Isoelectric focusing of the HeLa and hCG subunits demonstrated that the tumor protein had a lower pI. Immunoprecipitation and electrophoresis of ?-subunit from HeLa cultures labeled with [3H]fucose indicated that the tumor subunit was fucosylated, whereas analysis of hCG-? hydrosylates by HPLC confirmed previous reports that the placental subunit does not contain fucose. The results indicate that, regardless of whether or not a single ?-subunit gene is being expressed in both normal and neoplastic tissues, posttranslational modifications lead to a highly altered subunit in the tumor. The differences observed may be useful in diagnosing neoplastic vs hyperplastic conditions and may lend insight into the mechanism of ectopic hormone production by tumors

1988-08-23

227

Crosstalk with cancer-associated fibroblasts increases the growth and radiation survival of cervical cancer cells.  

Science.gov (United States)

Crosstalk between cancer cells and the surrounding cancer associated fibroblasts (CAFs) plays an illusive role in cancer radiotherapy. This study investigated the effect of cancer cell-cancer associated fibroblasts crosstalk on the proliferation and survival of irradiated cervical cancer cells. A pretreatment with conditioned medium from a mixed culture of CAF and HeLa cells (mixCAF) had a stronger effect on enhancing the proliferation and survival of irradiated HeLa cells compared to pretreatment with CAF conditioned medium alone. In addition, pretreatment with a mixed culture of CAF and HeLa cells conditioned medium reduced the levels of two major radiation-induced genes, GADD45 and BTG2, and phosphorylation of p38. Profiling of the growth and survival factors in the conditioned medium revealed PDGF and VEGF, and IGF2, EGF, FGF-4, IGFBPs and GM-CSF to be specifically secreted from HeLa cells and CAFs, respectively. This study demonstrated radiation protective effects of CAF-cancer cell crosstalk, and identified multiple growth factors and radiation response genes that might be involved in these effects. PMID:24785588

Chu, Tang-Yuan; Yang, June-Ting; Huang, Tien-Hung; Liu, Hwan-Wun

2014-05-01

228

Genome-wide profiling reveals a role for T-cell intracellular antigens TIA1 and TIAR in the control of translational specificity in HeLa cells.  

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TIA (T-cell intracellular antigens)-knockdown HeLa cells show an increase in ribosomes and translational machinery components. This increase correlates with specific changes in translationally up-regulated mRNAs involved in cell-cycle progression and DNA repair, as shown in polysomal profiling analysis. Our data support the hypothesis that a concerted activation of both global and selective translational rates leads to the transition to a more proliferative status in TIA-knockdown HeLa cells. PMID:24927121

Carrascoso, Isabel; Sánchez-Jiménez, Carmen; Izquierdo, José M

2014-07-01

229

Human papillomavirus 18 E6 inhibits phosphorylation of p53 expressed in HeLa cells  

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Full Text Available Abstract Background In HPV infected cells p53 function is abrogated by E6 and even ectopically expressed p53 is unable to perform tumor suppressor functions. In addition to facilitating its degradation, E6 may also inhibit p53 transactivity, though the mechanisms are still poorly understood. It has been reported that inhibition of p300, an acetyltransferase responsible for p53 acetylation is inactivated by E6. Activation of overexpressed p53 to cause cell growth inhibition is facilitated by its phosphorylation. Previously, we reported that non-genotoxically overexpressed p53 in HeLa cells needs to be phosphorylated to perform its cell growth inhibitory functions. Since over expressed p53 by itself was not activated, we hypothesized an inhibitory role for E6. Results Majority of reports proposes E6 mediated degradation of p53 as a possible reason for its inactivation. However, results presented here for the first time demonstrate that overexpressed p53 is not directly associated with E6 and therefore free, yet it is not functionally active in HPV positive cells. Also, the stability of overexpressed p53 does not seem to be an issue because inhibition of proteasomal degradation did not increase the half-life of overexpressed p53, which is more than endogenous p53. However, inhibition of proteasomal degradation prevents the degradation of endogenous p53. These findings suggest that overexpressed p53 and endogenous p53 are differentially subjected to proteasomal degradation and the reasons for this discrepancy remain unclear. Our studies demonstrate that p53 over expression has no effect on anchorage independent cell-growth and E6 nullifies its cell growth inhibitory effect. E6 overexpression abrogates OA induced p53 occupancy on the p21 promoter and cell death as well. E6 did not decrease p53 protein but phospho-p53 level was significantly reduced. Conclusion We report for the first time that E6 de-activates p53 by inhibiting its phosphorylation. This prevents p53 binding to p21 promoter and thereby restraining its cell-growth inhibitory functions. Our study provides new evidence indicating that viral protein E6 inhibits p53 transactivity by mechanism independent of degradation pathway.

Ajay Amrendra K

2012-01-01

230

Phospholipase C-?1 and ?4 Contribute to Non-Genetic Cell-to-Cell Variability in Histamine-Induced Calcium Signals in HeLa Cells  

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A uniform extracellular stimulus triggers cell-specific patterns of Ca2+ signals, even in genetically identical cell populations. However, the underlying mechanism that generates the cell-to-cell variability remains unknown. We monitored cytosolic inositol 1,4,5-trisphosphate (IP3) concentration changes using a fluorescent IP3 sensor in single HeLa cells showing different patterns of histamine-induced Ca2+ oscillations in terms of the time constant of Ca2+ spike amplitude decay and the Ca2+ oscillation frequency. HeLa cells stimulated with histamine exhibited a considerable variation in the temporal pattern of Ca2+ signals and we found that there were cell-specific IP3 dynamics depending on the patterns of Ca2+ signals. RT-PCR and western blot analyses showed that phospholipase C (PLC)-?1, -?3, -?4, -?1, -?3 and -? were expressed at relatively high levels in HeLa cells. Small interfering RNA-mediated silencing of PLC isozymes revealed that PLC-?1 and PLC-?4 were specifically involved in the histamine-induced IP3 increases in HeLa cells. Modulation of IP3 dynamics by knockdown or overexpression of the isozymes PLC-?1 and PLC-?4 resulted in specific changes in the characteristics of Ca2+ oscillations, such as the time constant of the temporal changes in the Ca2+ spike amplitude and the Ca2+ oscillation frequency, within the range of the cell-to-cell variability found in wild-type cell populations. These findings indicate that the heterogeneity in the process of IP3 production, rather than IP3-induced Ca2+ release, can cause cell-to-cell variability in the patterns of Ca2+ signals and that PLC-?1 and PLC-?4 contribute to generate cell-specific Ca2+ signals evoked by G protein-coupled receptor stimulation.

Ishida, Sachiko; Matsu-ura, Toru; Fukami, Kiyoko; Michikawa, Takayuki; Mikoshiba, Katsuhiko

2014-01-01

231

Quantification of Functionalised Gold Nanoparticle-Targeted Knockdown of Gene Expression in HeLa Cells  

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Introduction Gene therapy continues to grow as an important area of research, primarily because of its potential in the treatment of disease. One significant area where there is a need for better understanding is in improving the efficiency of oligonucleotide delivery to the cell and indeed, following delivery, the characterization of the effects on the cell. Methods In this report, we compare different transfection reagents as delivery vehicles for gold nanoparticles functionalized with DNA oligonucleotides, and quantify their relative transfection efficiencies. The inhibitory properties of small interfering RNA (siRNA), single-stranded RNA (ssRNA) and single-stranded DNA (ssDNA) sequences targeted to human metallothionein hMT-IIa are also quantified in HeLa cells. Techniques used in this study include fluorescence and confocal microscopy, qPCR and Western analysis. Findings We show that the use of transfection reagents does significantly increase nanoparticle transfection efficiencies. Furthermore, siRNA, ssRNA and ssDNA sequences all have comparable inhibitory properties to ssDNA sequences immobilized onto gold nanoparticles. We also show that functionalized gold nanoparticles can co-localize with autophagosomes and illustrate other factors that can affect data collection and interpretation when performing studies with functionalized nanoparticles. Conclusions The desired outcome for biological knockdown studies is the efficient reduction of a specific target; which we demonstrate by using ssDNA inhibitory sequences targeted to human metallothionein IIa gene transcripts that result in the knockdown of both the mRNA transcript and the target protein.

Jiwaji, Meesbah; Sandison, Mairi E.; Reboud, Julien; Stevenson, Ross; Daly, Ronan; Barkess, Grainne; Faulds, Karen; Kolch, Walter; Graham, Duncan; Girolami, Mark A.; Cooper, Jonathan M.; Pitt, Andrew R.

2014-01-01

232

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

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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

233

3D printing of biomimetic microstructures for cancer cell migration.  

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To understand the physical behavior and migration of cancer cells, a 3D in vitro micro-chip in hydrogel was created using 3D projection printing. The micro-chip has a honeycomb branched structure, aiming to mimic 3D vascular morphology to test, monitor, and analyze differences in the behavior of cancer cells (i.e. HeLa) vs. non-cancerous cell lines (i.e. 10 T1/2). The 3D Projection Printing system can fabricate complex structures in seconds from user-created designs. The fabricated microstructures have three different channel widths of 25, 45, and 120 microns wide to reflect a range of blood vessel diameters. HeLa and 10 T1/2 cells seeded within the micro-chip were then analyzed for morphology and cell migration speed. 10 T1/2 cells exhibited greater changes in morphology due to channel size width than HeLa cells; however, channel width had a limited effect on 10 T1/2 cell migration while HeLa cancer cell migration increased as channel width decreased. This physiologically relevant 3D cancer tissue model has the potential to be a powerful tool for future drug discoveries and cancer migration studies. PMID:24150602

Huang, Tina Qing; Qu, Xin; Liu, Justin; Chen, Shaochen

2014-02-01

234

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

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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

235

Specific interactions of HeLa cell proteins with proposed translation domains of the poliovirus 5' noncoding region.  

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To determine which sequences or structures in the poliovirus 5' noncoding region (5'NCR) are involved in binding proteins used for internal ribosome binding and protein synthesis initiation, translation competition assays were performed in rabbit reticulocyte lysates in the presence and absence of HeLa cell extract. The results revealed two functional domains in the poliovirus 5'NCR. One, requiring nucleotides (nts) 457 to 626, binds proteins that are required for translation of all mRNAs and...

Gebhard, J. R.; Ehrenfeld, E.

1992-01-01

236

Localization of HeLa cell tumor-suppressor gene to the long arm of chromosome II.  

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Cytogenetic and molecular genetic analyses of human intraspecific HeLa x fibroblast hybrids have provided evidence for the presence of a tumor-suppressor gene(s) on chromosome 11 of normal cells. In the present study, we have carried out extensive RFLP analysis of various nontumorigenic and tumorigenic hybrids with at least 50 different chromosome 11-specific probes to determine the precise location of this tumor-suppressor gene(s). Two different hybrid systems, (1) microcell hybrids derived ...

Misra, B. C.; Srivatsan, E. S.

1989-01-01

237

The Continuous Passage of Agents of Trachoma in Cell Culture. I. Characteristics of Tw-3 and Bour Strains of Trachoma Cultivated in Serial Passage in Hela 229 Cells.  

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Twelve strains and lines of mammalian cells were screened for their susceptibility to infection with the TW-3 and Bour strains of trachoma. The HeLa 229 cell line showed the highest susceptibility to infection. At no time were all the cells infected nor w...

H. M. Jenkin

1966-01-01

238

Cytotoxic activity of proteins isolated from extracts of Corydalis cava tubers in human cervical carcinoma HeLa cells  

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Full Text Available Abstract Background Corydalis cava Schweigg. & Koerte, the plant of numerous pharmacological activities, together with the studied earlier by our group Chelidonium majus L. (Greater Celandine, belong to the family Papaveraceae. The plant grows in Central and South Europe and produces the sizeable subterraneous tubers, empty inside, which are extremely resistant to various pathogen attacks. The Corydalis sp. tubers are a rich source of many biologically active substances, with the extensive use in European and Asian folk medicine. They have analgetic, sedating, narcotic, anti-inflammatory, anti-allergic and anti-tumour activities. On the other hand, there is no information about possible biological activities of proteins contained in Corydalis cava tubers. Methods Nucleolytic proteins were isolated from the tubers of C. cava by separation on a heparin column and tested for DNase activity. Protein fractions showing nucleolytic activity were tested for cytotoxic activity in human cervical carcinoma HeLa cells. Cultures of HeLa cells were conducted in the presence of three protein concentrations: 42, 83 and 167 ng/ml during 48 h. Viability of cell cultures was appraised using XTT colorimetric test. Protein fractions were separated and protein bands were excised and sent for identification by mass spectrometry (LC-ESI-MS/MS. Results The studied protein fractions showed an inhibiting effect on mitochondrial activity of HeLa cells, depending on the administered dose of proteins. The most pronounced effect was obtained with the highest concentration of the protein (167 ng/ml - 43.45 ± 3% mitochondrial activity of HeLa cells were inhibited. Mass spectrometry results for the proteins of applied fractions showed that they contained plant defense- and pathogenesis-related (PR proteins. Conclusions The cytotoxic effect of studied proteins toward HeLa cell line cells has been evident and dependent on increasing dose of the protein. The present study, most probably, represents the first investigations on the effect of purified PR proteins from tuber extracts of a pharmacologically active plant on cell lines.

Balcerkiewicz Stanislaw

2010-12-01

239

Protective effect of mitochondrial ferritin on cytosolic iron dysregulation induced by doxorubicin in HeLa cells.  

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Doxorubicin (DOX) is an anticancer drug with cardiotoxic side effects mostly caused by iron homeostasis dysregulation. Mitochondria are involved in iron trafficking and mitochondrial ferritin (FtMt) was shown to provide protection against cellular iron imbalance. Therefore, we hypothesized that FtMt overexpression could limit DOX effects on iron homeostasis. Heart's homogenates of DOX-treated C57BL/6 mice were analyzed for cytosolic and mitochondrial iron-related proteins' expression and activity, revealing high cytosolic ferritin and ferritin-bound iron, low transferrin-receptor 1 and a strong hepcidin upregulation. Mitochondrial iron-related proteins (aconitase, succinate-dehydrogenase, frataxin) seemed, however, unaffected, although a partial inactivation of superoxide dismutase 2 was detected. Importantly, the ectopic expression of FtMt in human HeLa cells partially reverted DOX-induced iron imbalance. Our results, while confirming DOX effects on iron homeostasis, demonstrate that DOX affects more cytosolic than mitochondrial iron metabolism both in murine hearts and human HeLa cells and that FtMt overexpression is able to prevent most of these effects in HeLa cells. PMID:24065548

Cocco, Emiliano; Porrini, Vanessa; Derosas, Manuela; Nardi, Veronica; Biasiotto, Giorgio; Maccarinelli, Federica; Zanella, Isabella

2013-12-01

240

In vitro activity of nonoxynol 9 on HeLa 229 cells and primary monkey cervical epithelial cells infected with Chlamydia trachomatis.  

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Nonoxynol 9 (non-9) is the active ingredient in a wide variety of vaginal contraceptive preparations. The manufacturer recommendation for optimal contraceptive practice is repeated application every 6 h. We studied the in vitro activity of non-9 against Chlamydia trachomatis (E/UW-5/Cx) and its toxicity against HeLa 229 cells and monkey cervical epithelial cells. With a contact time of 6 h, non-9 was toxic to HeLa cells at concentrations of 50 micrograms/ml or greater and to monkey cervical c...

1992-01-01

 
 
 
 
241

Highly sensitive determination of copper in HeLa cell using capillary electrophoresis combined with a simple cell extraction treatment.  

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A new separation system of capillary electrophoresis (CE1) for the highly sensitive determination of copper was established by using ethylenediaminetetraacetic acid (EDTA) as a complexing agent and employing cetyltrimethylammonium chloride (CTAC) as a capillary inner wall modifier. Benefitted from the combination of field-enhanced sample injection (FESI) method, a limit of detection (LOD) of 2.7 nM was obtained, which was much lower than that of the conventional methods. This made it possible to determine trace copper in HeLa cell only by a simple cell extraction (CE2) treatment. Two copper-extraction methods-acid-hydrolysis and freeze-thaw-were compared. Limited by the requirement of low ion strength in FESI, only the extract using freeze-thaw could be successfully applied in the determination. The effectiveness assessment of this CE(2)-FESI method was adopted by inductively coupled plasma-atomic emission spectrometry (ICP-AES) as a gold standard. PMID:24607128

Meng, Lingchen; Fang, Ziyuan; Lin, Jian; Li, Meixian; Zhu, Zhiwei

2014-04-01

242

Diarylheptanoid-myricanone isolated from ethanolic extract of Myrica cerifera shows anticancer effects on HeLa and PC3 cell lines: signalling pathway and drug-DNA interaction  

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Full Text Available OBJECTIVE: To test if myricanone (C21H24O5, a cyclic diarylheptanoid, has anticancer effects on two different cancer cell lines HeLa and PC3. The present study was conducted with a note on the drug-DNA interaction and apoptotic signalling pathway.METHODS: Several studies like cytotoxicity, nuclear damage, annexin-V-fluorescein isothiocyanate (FITC/propidium iodide (PI-labelled apoptotic assay and cell cycle arrest, immunoblot and reverse transcriptase-polymerase chain reaction (RT-PCR were used following standard protocols. Circular dichroism (CD spectroscopy was also done to evaluate whether myricanone effectively interacted with DNA to bring about conformational changes that could strongly inhibit the cancer cell proliferation.RESULTS: Myricanone showed a greater cytotoxic effect on PC3 cells than on HeLa cells. Myricanone promoted G0/G1 arrest in HeLa cells and S phase arrest in PC3 cells. Nuclear condensation and annexin V-FITC/PI studies revealed that myricanone promoted apoptotic cell death. CD spectroscopic data indicated that myricanone had an interaction with calf thymus DNA that changed DNA structural conformation. RT-PCR and immunoblot studies revealed that myricanone activated the apoptotic signalling cascades through down-regulation of transcription factors like nuclear factor-?B (NF-?B (p65, and signal transducers and activators of transcription 3 (STAT3; cell cycle regulators like cyclin D1, and survivin and other signal proteins like Bcl-2 and up-regulation of Bax, caspase-9 and caspase-3.CONCLUSION: Myricanone induced apoptosis in both types of cancer cells by triggering caspase activation, and suppression of cell proliferation by down-regulation of NF-?B and STAT3 signalling cascades, which makes it a suitable candidate for possible use in the formulation of therapeutic agent for combating cancer.

Avijit Paul

2013-11-01

243

Identification of genes associated to 2',2'-difluorodeoxycytidine resistance in HeLa cells with a lentiviral short-hairpin RNA library.  

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Resistance to the cytotoxic nucleoside analog 2',2'-diflurodeoxycytidine (dFdC) used in cancer chemotherapy is a frequent cause of treatment failure. Although several molecular mechanisms that cause resistance to dFdC have been identified, many cells acquire dFdC resistance by unknown mechanisms. We have used a short-hairpin RNA (shRNA) library in a lentiviral vector that contains ?5000 shRNAs designed against genes encoding kinases, phosphatases, tumor suppressors and DNA binding proteins to perform a loss-of-function screen to identify genes causing dFdC resistance in HeLa cells when their expression is decreased. 155 cell lines with shRNA expression were isolated from the screen and several of these cell lines were in repeated experiments verified to show resistance to dFdC compared to wild-type cells. DNA sequencing of the shRNA vector integrated in the cellular genome was used to determine the shRNA expressed in the cells and the putative target genes were identified by sequence analysis. 16 cell lines with putative target genes previously not associated to dFdC resistance were identified. Chemically synthesized short-interfering RNAs (siRNAs) directed against the target genes were used to verify that the decreased expression of the identified genes caused dFdC resistance. Using these techniques we identified two splicing factor proteins, serine/arginine-rich splicing factor 3 (SRSF3) and splicing factor proline/glutamine-rich (SFPQ), that induced resistance to dFdC as well as other pyrimidine nucleoside analogs when their expression was decreased in HeLa cells. PMID:21565176

Xu, Yunjian; Karlsson, Anna; Johansson, Magnus

2011-08-01

244

Effect of polyamine depletion on DNA damage and repair following UV irradiation of HeLa cells  

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Treatment of HeLa cells with the polyamine biosynthesis inhibitors, methylglyoxal bis(guanylhydrazone) (MGBG), difluoromethylornithine (DFMO) or a combination of the two, resulted in reduction in cellular polyamine levels. Analysis of UV light-induced DNA damage and repair in these polyamine depleted cells revealed distinct differences in the repair process relative to that seen in cells possessing a normal polyamine complement. Observed patterns of differential polyamine depletion by DFMO and MGBG, and partial reversal of repair inhibition by polyamine supplementation, suggest that polyamine depletion per se, rather than some secondary effect of inhibitor treatment, is responsible for the inhibition of repair. (author).

Snyder, R.D.; Sunkara, P.S. (Merrell Dow Research Inst., Cincinnati, OH (USA))

1990-09-01

245

Effect of polyamine depletion on DNA damage and repair following UV irradiation of HeLa cells  

International Nuclear Information System (INIS)

Treatment of HeLa cells with the polyamine biosynthesis inhibitors, methylglyoxal bis(guanylhydrazone) (MGBG), difluoromethylornithine (DFMO) or a combination of the two, resulted in reduction in cellular polyamine levels. Analysis of UV light-induced DNA damage and repair in these polyamine depleted cells revealed distinct differences in the repair process relative to that seen in cells possessing a normal polyamine complement. Observed patterns of differential polyamine depletion by DFMO and MGBG, and partial reversal of repair inhibition by polyamine supplementation, suggest that polyamine depletion per se, rather than some secondary effect of inhibitor treatment, is responsible for the inhibition of repair. (author)

1990-01-01

246

Sulfated fucan from marine alga inhibits HeLa cells infection by HTLV-1 free particles: semi-quantitative analysis  

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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 when added to the cel...

Romanos, Maria T. V.; Andrada-serpa, Maria J.; Moura?o, Paulo A. S.; Yocie Yoneshigue-Valentin; Pereira, Mariana S.; Norma Santos; Wigg, Marcia D.

2011-01-01

247

Filament Tip-Associated Antigens Involved in Adherence to and Invasion of Murine Pulmonary Epithelial Cells In Vivo and HeLa Cells In Vitro by Nocardia asteroides  

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The interactions of Nocardia asteroides GUH-2 with pulmonary epithelial cells of C57BL/6 mice and with HeLa cells were studied. Electron microscopy demonstrated that only the tips of log-phase cells penetrated pulmonary epithelial cells following intranasal administration, and nocardiae were recovered from the brain. Coccobacillary cells neither invaded nor disseminated. Serum from immunized mice (IMS) decreased attachment to and penetration of pulmonary epithelial cell surfaces by log-phase ...

Beaman, Blaine L.; Beaman, Lovelle

1998-01-01

248

Down-regulation of the sodium pump following chronic exposure of HeLa cells and chick embryo heart cells to ouabain.  

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1 HeLa cells and primary cultures of embryonic chick heart cells were grown in medium containing low concentrations of ouabain for 24 h. 2 Compared with normal cells, cells grown in ouabain have fewer free sodium pump sites, an increased intracellular sodium concentration and a decreased intracellular potassium concentration. The cells are able to maintain their intracellular ion contents because the remaining pump sites have an increased turnover rate. 3 When cells that have been chronically...

Aiton, J. F.; Lamb, J. F.; Ogden, P

1981-01-01

249

Control of placental alkaline phosphatase gene expression in HeLa cells: induction of synthesis by prednisolone and sodium butyrate  

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HeLa S3 cells produce an alkaline phosphatase indistinguishable from the enzyme from human term placenta. The phosphatase activity in these cells was induced by both prednisolone and sodium butyrate. Both agents stimulated de novo synthesis of the enzyme. The increase in phosphatase activity paralleled the increase in immunoactivity and biosynthesis of placental alkaline phosphatase. The fully processed phosphatase monomer in control, prednisolone-treated or butyrate-treated cells was a 64.5 K polypeptide, measured by both incorporation of L-[35S]methionine into enzyme protein and active-site labeling. The 64.5K polypeptide was formed by the incorporation of additional N-acetylneuraminic acid moieties to a precursor polypeptide of 61.5K. However, this biosynthetic pathway was identified only in butyrate-treated cells. In prednisolone-treated cells, the processing of 61.5K to 64.5K monomer was accelerated, and the presence of the 61.5 precursor could only be detected by either neuraminidase or monensin treatment. Phosphatase mRNA which comigrated with the term placental alkaline phosphatase mRNA of 2.7 kilobases was induced in the presence of either prednisolone or butyrate. Alkaline phosphatase mRNA is untreated HeLa S3 cells migrated slightly faster than the term placental alkaline phosphatase mRNA. Butyrate also induced a second still faster migrating alkaline phosphatase mRNA. Both prednisolone and butyrate increased the steady-state levels of placental alkaline phosphatase mRNA. The data indicate that the increase in phosphatase mRNA by prednisolone and butyrate resulted in the induction of alkaline phosphatase activity and biosynthesis in HeLa S3 cells. Furthermore, both agents induced the expression of different alkaline phosphatase gene transcripts without altering its protein product

1987-06-16

250

Nucleases isolated from Chelidonium majus L. milky sap can induce apoptosis in human cervical carcinoma HeLa cells but not in Chinese Hamster Ovary CHO cells.  

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Full Text Available Milky sap isolated from Chelidonium majus L. (Greater Celandine serves as a rich source of various biologically active substances such as alkaloids, flavonoids and phenolic acids. Previous research showed that the activity of Ch. majus milky sap may depend also on the presence of biologically active proteins. The goal of this study was to evaluate the biological effect of two nucleases isolated from Ch. majus milk sap, CMN1 of 20 kDa and CMN2 of 36 kDa, on HeLa and CHO tumour cell lines. Both studied nucleases together with other proteins in the sap of the plant are involved in stress and defence reactions against different pathogens. After 48 h incubation of CMN1 and CMN2 only with HeLa cells, the dependence between the number of apoptotic lesions and the concentration of applied nuclease was observed. The highest proapoptotic activity was induced by 13.3 ng/ml concentration of CMN2 collected in May (62 +/- 3% HeLa cells were apoptotic. Moreover, the proportion of necrotic cells in all concentrations of the nucleases and both cell lines was relatively low (1-8 +/- 0.5%. In summary, results of this study show that purified nucleases CMN1 and CMN2 isolated from Ch. majus milky sap exhibit apoptotic activity in HeLa tumour cell line, but not in CHO cells, without inflammatory reaction.

Maria Wo?u?-Cholewa

2008-02-01

251

ADP-ribosylation of nonhistone proteins from metaphase and interphase HeLa cells: factors responsible for differences  

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A striking reduction was previously detected for HeLa metaphase chromosomes, compared to interphase nuclei, in the number of modified nonhistone species. Several factors which could contribute to this cell cycle change in ADP-ribosylation have therefore been examined. In these experiments, mitotic or interphase cells were incubated with ["3"2P]NAD, chromosomes and nuclei were prepared, and the proteins were resolved by polyacrylamide gel electrophoresis. The level of incorporation of "3"2P label was found to be substantially influenced by chromosome expansion, DNA nicking, disruption of chromosomes or nuclei, and the growth activity of cells. The level of ADP-ribosylation was not greatly affected by the presence of inhibitors of RNA, DNA, and protein synthesis. NAD concentration influenced the extent of labelling but not the pattern of labeled species. A similar change in the pattern from interphase to mitosis was observed for whole cells as well as for isolated chromosomes and nuclei. The procedure used to arrest cells in mitosis was not artifactually responsible for the results. The difference in metaphase and interphase ADP-ribosylation is not confined to HeLa cells, since comparable patterns were found for chromosomes and nuclei from Novikoff rat hepatoma cells

1986-05-01

252

Synthesis and methylation of ribosomal RNA in HeLa cells infected with the herpes virus pseudorabies virus  

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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

253

Synthesis and methylation of ribosomal RNA in HeLa cells infected with the herpes virus pseudorabies virus  

International Nuclear Information System (INIS)

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

254

Intercellular Calcium Waves in HeLa Cells Expressing GFP-labeled Connexin 43, 32, or 26  

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This study was undertaken to obtain direct evidence for the involvement of gap junctions in the propagation of intercellular Ca2+ waves. Gap junction-deficient HeLa cells were transfected with plasmids encoding for green fluorescent protein (GFP) fused to the cytoplasmic carboxyl termini of connexin 43 (Cx43), 32 (Cx32), or 26 (Cx26). The subsequently expressed GFP-labeled gap junctions rendered the cells dye- and electrically coupled and were detected at the plasma membranes at points of con...

Paemeleire, Koen; Martin, Patricia E. M.; Coleman, Sharon L.; Fogarty, Kevin E.; Carrington, Walter A.; Leybaert, Luc; Tuft, Richard A.; Evans, W. Howard; Sanderson, Michael J.

2000-01-01

255

Development of electrochemical reporter assay using HeLa cells transfected with vector plasmids encoding various responsive elements  

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Electrochemical assay using HeLa cell lines transfected with various plasmid vectors encoding SEAP (secreted alkaline phosphatase) as the reporter has been performed by using SECM (scanning electrochemical microscopy). The plasmid vector contains different responsive elements that include GRE (glucocorticoid response elements), CRE (cAMP responsive elements), or {kappa}B (binding site for NF{kappa}B (nuclear factor kappa B)) upstream of the SEAP sequence. The transfected HeLa cells were patterned on a culture dish in a 4 x 4 array of circles of diameter 300 {mu}m by using the PDMS (poly(dimethylsiloxane)) stencil technique. The cellular array was first exposed to 100 ng mL{sup -1} dexamethasone, 10 ng mL{sup -1} forskolin, or 100 ng mL{sup -1} TNF-{alpha} (tumor necrosis factor {alpha}) after which it was further cultured in an RPMI culture medium for 6 h. After incubation, the cellular array was soaked in a measuring solution containing 4.7 mM PAPP (p-aminophenylphosphate) at pH 9.5, following which electrochemical measurements were performed immediately within 40 min. The SECM method allows parallel evaluation of different cell lines transfected with pGRE-SEAP, pCRE-SEAP, and pNF{kappa}B-SEAP patterned on the same solid support for detection of the oxidation current of PAP (p-aminophenol) flux produced from only 300 HeLa cells in each stencil pattern. The results of the SECM method were highly sensitive as compared to those obtained from the conventional CL (chemiluminescence) protocol with at least 5 x 10{sup 4} cells per well.

Shiku, Hitoshi, E-mail: shiku@bioinfo.che.tohoku.ac.jp [Graduate School of Environmental Studies, Tohoku University, 6-6-11-604 Aramaki-Aoba, Sendai 980-8579 (Japan); Takeda, Michiaki; Murata, Tatsuya [Graduate School of Environmental Studies, Tohoku University, 6-6-11-604 Aramaki-Aoba, Sendai 980-8579 (Japan); Akiba, Uichi; Hamada, Fumio [Graduate School of Engineering and Resource Science, Akita University, 1-1 Tegata gakuen-machi, Akita 010-8502 (Japan); Matsue, Tomokazu, E-mail: matsue@bioinfo.che.tohoku.ac.jp [Graduate School of Environmental Studies, Tohoku University, 6-6-11-604 Aramaki-Aoba, Sendai 980-8579 (Japan)

2009-04-27

256

Interaction of hyperthermia and radiation in tolerant and nontolerant HeLa S3 cells: role of DNA polymerase inactivation  

International Nuclear Information System (INIS)

The activities of DNA polymerase ? and ? were measured in tolerant and nontolerant HeLa S3 suspension cells. The heat-inactivation of the enzymes and their recovery when cells were incubated at 370C after the heat challenge was compared to the synergistic action of heat and x-radiation and its disappearance at the level of cell survival. Thermotolerant cells were radiosensitized by heat similarly to nontolerant cells, but the sensitization decreased more rapidly in the tolerant cells when time at 370C was allowed between the two treatments. For polymerase activities the extent of inactivation, as well as the kinetics of recovery, were similar in tolerant and nontolerant cells. (author)

1989-01-01

257

X-ray microanalysis of HeLa S3 Cells. I. Instrumental calibration and analysis of randomly growing cultures.  

Science.gov (United States)

Cryo-ultramicrotomy and X-ray microanalysis were used to study the elemental composition of HeLa S3 cells. Quantitation was achieved by reference to elemental standards of known concentration made up in 25% gelatin. Analysis of standards showed linear calibration for each of the elements studied: Na, P, S, Cl, K. Standardization was validated by comparing flame-photometric analysis of gelatin containing sodium potassium tartrate with that of X-ray microanalysis. Freeze-dried sections of cells showed good morphology and analysis of whole sections of the cells showed that K/Na varied in individual cells. Low K/Na could not be ascribed to cell damage or to the sequestering of Na in any particular subcompartment of the cells. Treatment with ouabain caused changes in levels of all the elements studied and resulted in a low K/Na ratio in all cells. PMID:6874730

Warley, A; Stephen, J; Hockaday, A; Appleton, T C

1983-03-01

258

Pronounced transcriptional regulation of apoptotic and TNF-NF-kappa-B signaling genes during the course of thymoquinone mediated apoptosis in HeLa cells.  

Science.gov (United States)

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. PMID:23943306

Sakalar, Cagri; Yuruk, Merve; Kaya, Tugba; Aytekin, Metin; Kuk, Salih; Canatan, Halit

2013-11-01

259

Expression of aggregative adherence to hela cells by Escherichia coli strains isolated from sick horses Expressão de aderência agregativa em células HeLa por amostras de E. coli isoladas de eqüinos doentes  

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The virulence attributes of 56 Escherichia coli strains isolated from sick horses (secretions of uterine cervices; gastrointestinal and lung fragments of necropsy; diarrheic feces, and tracheal washings) was examined by determining their adherence pattern to HeLa cells and searching for the presence of virulence genes of the various E. coli pathotypes. Two non-adherent strains presented astA, which encodes the enteroaggregative E. coli heat-stable toxin. Twenty-seven strains (48.2%) adhered t...

Ana Maria Alvim Liberatore; Sandra Kimie Tomita; Mônica Aparecida Midolli Vieira; Cyro Toti Jr.; João Heckmaier; Tânia Aparecida Tardelli Gomes

2007-01-01

260

A prominent low-pH methotrexate transport activity in human solid tumors: contribution to the preservation of methotrexate pharmacologic activity in HeLa cells lacking the reduced folate carrier.  

Science.gov (United States)

Whereas the major folate transporter, the reduced folate carrier (RFC), has a physiological pH optimum, transport activities for folates and antifolates have been detected with low pH optima. Because the interstitial pH in solid tumors is generally acidic, the mechanisms by which antifolates are transported at low pH could be an important determinant of drug activity under these conditions. The current study quantitated the low pH methotrexate (MTX) transport activity in human solid tumor cell lines from the National Cancer Institute tumor panel and other sources. MTX influx at pH 5.5 was equal to, or greater than, influx at pH 7.4 in 29 of 32 cell lines. To assess the role of RFC in transport at low pH in one of these cell lines, a HeLa clonal line (R5) was selected for MTX resistance due to a genomic deletion of the carrier gene. MTX influx was depressed by 70% in R5 versus wild-type HeLa cells at pH 7.4. At pH 6.5, influx in these two lines was similar; as the pH was decreased to 5.5 influx increased in both cell lines. Similarly, whereas net MTX uptake over 1 h was markedly decreased in R5 cells at pH 7.4, net uptake in HeLa and R5 cells was comparable at pH 6.5. Also, as compared with MCF7 breast cancer cells, MTX uptake was markedly decreased at pH 7.4, but only minimally at pH 6.5, in the MDA-MB-231 human breast cancer cell line that lacks RFC expression. When grown with folic acid (2 micro M) at pH 7.4, the IC(50) for MTX was 14-fold higher in R5 as compared with wild-type HeLa cells; the difference was only 4-fold when cells when grown at pH 6.9; the IC(50)s were identical at this pH when the medium folate was 25 nM 5-formyltetrahydrofolate. These data demonstrate that transport activity at low pH is prevalent in human solid tumors, is RFC-independent in R5 cells and MDA-MB-231 breast cancer cells, and can preserve MTX activity in the absence of RFC at an acidic pH relevant to solid tumors in vivo. PMID:14760095

Zhao, Rongbao; Gao, Feng; Hanscom, Marie; Goldman, I David

2004-01-15

 
 
 
 
261

SPONTANEOUS AND MNNG-INDUCED REVERSION OF AN EGFP CONSTRUCT IN HELA CELLS: AN ASSAY FOR OBSERVING MUTATIONS IN LIVING CELLS BY FLUORESCENT MICROSCOPY  

Science.gov (United States)

A HeLa cell line stably expressing the Enhanced Green Fluorescence Protein (EGFP) gene, interrupted by the IVS2-654 intron, was studied without treatment and after treatment with a single standard dose of 15 ?M of N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). This assay was done ...

262

Inhibition of X-ray induced DNA strand break repair in polyamine-depleted HeLa cells  

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Treatment of HeLa cells with the polyamine biosynthesis inhibitors, alpha-difluoromethylornithine (DFMO) or methylglyoxal bis(guanylhydrazone) (MGBG), results in, depending on the conditions, partial or complete depletion of the cellular polyamines: putrescine, spermidine and spermine. In this compromised state cells exhibited a distinct deficiency in repair of X-ray-induced DNA strand breaks. The half-time for return of normal DNA sedimentation following 1.6 Gy was 9.5 min for untreated control cells and 22, 32 and 50 min for cells treated with MGBG, DFMO+MGBG and DFMO, respectively. Normal repair kinetics were restored to these cells upon a short incubation in media containing all three polyamines. The rapid early phase of repair following higher X-ray doses (16 Gy) was also delayed in polyamine-depleted cells but later repair occurring 1-4 h post-irradiation, representing chromatin reconstitution, was apparently normal. (author).

Snyder, R.D.

1989-05-01

263

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 SciELO Brazil | Language: English 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 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..

264

Sulfated fucan from marine alga inhibits HeLa cells infection by HTLV-1 free particles: semi-quantitative analysis  

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Full Text Available 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 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.

Maria T. V. Romanos

2011-04-01

265

Inhibition of X-ray induced DNA strand break repair in polyamine-depleted HeLa cells  

International Nuclear Information System (INIS)

Treatment of HeLa cells with the polyamine biosynthesis inhibitors, alpha-difluoromethylornithine (DFMO) or methylglyoxal bis(guanylhydrazone) (MGBG), results in, depending on the conditions, partial or complete depletion of the cellular polyamines: putrescine, spermidine and spermine. In this compromised state cells exhibited a distinct deficiency in repair of X-ray-induced DNA strand breaks. The half-time for return of normal DNA sedimentation following 1.6 Gy was 9.5 min for untreated control cells and 22, 32 and 50 min for cells treated with MGBG, DFMO+MGBG and DFMO, respectively. Normal repair kinetics were restored to these cells upon a short incubation in media containing all three polyamines. The rapid early phase of repair following higher X-ray doses (16 Gy) was also delayed in polyamine-depleted cells but later repair occurring 1-4 h post-irradiation, representing chromatin reconstitution, was apparently normal. (author)

1989-01-01

266

Global gene expression profiling of HeLa and HepG2 cells in response to simulated microgravity  

Science.gov (United States)

Microgravity is considered a major environmental factor that affects cells and tissues causing adverse effects to human health during space flight Ground-based gravity simulation experiments at the cellular and molecular levels have given much insight into the underlying molecular and cellular alterations induced by microgravity stress In the present study we investigated microgravity effects on human cell lines such as HeLa cells and HepG2 cells under simulated microgravity conditions using the Rotating Wall Vessel Bioreactor Gene expression profiles of time course microgravity treated cells were displayed through DNA microarray analysis Some of the microgravity-responsive genes were further validated using Northern and RT-PCR techniques The identified set of genes that are preferentially altered in microgravity conditions may constitute part of the major space genes that together play a major check-and-balance role ultimately determining the outcome of a cell or an organism in response to microgravity conditions

Clement, J.; Lacy, S.; Wilson, B.

267

Lactoferrin and free secretory component of human milk inhibit the adhesion of enteropathogenic Escherichia coli to HeLa cells  

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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.

Giugliano Loreny

2001-10-01

268

Adhesion of Escherichia coli to HeLa Cells Mediated by Trypanosoma cruzi Surface Glycoprotein-Derived Peptides Inserted in the Outer Membrane Protein LamB  

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Peptides derived from the surface glycoprotein gp82 of Trypanosoma cruzi, previously implicated in the parasite’s invasion of host cells, were expressed as fusions to the protein LamB of Escherichia coli in a region known to be exposed on the cell surface. Bacteria expressing these proteins adhered to HeLa cells in a manner that mimics the pattern of parasite invasion of mammalian cells. Purified LamB fusion proteins were shown to bind to HeLa cells and to inhibit infection by T. cruzi, sup...

Pereira, Ca?tia M.; Favoreto, Si?lvio; Franco Da Silveira, Jose?; Yoshida, Nobuko; Castilho, Beatriz A.

1999-01-01

269

Medium from X-rayed cultures induces DNA strand-breaks in non-irradiated HeLa cells  

International Nuclear Information System (INIS)

There is growing evidence to indicate that several types of responses are induced by ionizing radiation in non-irradiated cells. Such bystander effects include the killing of non-irradiated cells, the induction of sister chromatid exchanges and chromosomal aberrations, and the induction of gene mutations and chromosomal instability and enhanced cell growth. In the present study, we assessed whether the medium from irradiated cultures can induce DNA strand-breaks in non-irradiated cells, using single-cell gel electrophoresis assay (comet assay). HeLa cells in culture were irradiated with 0.5 to 8 Gy of 140 kVp X-rays and one hour later, the medium was taken from the irradiated culture, passed through a filter and transferred to the parallel culture of non-irradiated HeLa cells as non-target cells. After incubation for 30 min, the comet assay was performed under alkaline and neutral conditions. Such treatments resulted in a dose-dependent increase in tail moment under either alkaline or neutral condition, indicating the induction of DNA single- or double-strand breaks, respectively. It was also shown that the clonogenic survival was reduced in the cells cultured in the medium from irradiated cultures. Such a change was not detected at all when medium alone was irradiated. These results provided disputed evidence that irradiated cells released certain genotoxic factor(s) into the culture medium that can induce DNA strand breaks leading to cell death. Our results suggest that physical contact between irradiated and non-irradiated cells may not be necessary for the bystander effects observed in this study. It appears that bystander responses may be mediated by multiple mechanisms

2002-10-20

270

Anomalous diffusion of major histocompatibility complex class I molecules on HeLa cells determined by single particle tracking.  

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Single-particle tracking (SPT) was used to determine the mobility characteristics of MHC (major histocompatibility complex) class I molecules at the surface of HeLa cells at 22 degrees C and on different time scales. MHC class I was labeled using the Fab fragment of a monoclonal antibody (W6/32), covalently bound to either R-phycoerythrin or fluorescent microspheres, and the particles were tracked using high-sensitivity fluorescence imaging. Analysis of the data for a fixed time interval sugg...

Smith, P. R.; Morrison, I. E.; Wilson, K. M.; Ferna?ndez, N.; Cherry, R. J.

1999-01-01

271

Inhibition of Chlamydia trachomatis growth in McCoy, HeLa, and human prostate cells by zinc.  

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Zinc salts (10(-4) and 10(-5) M) inhibited the number of Chlamydia trachomatis inclusions in McCoy, HeLa, and primary human prostate epithelial cell cultures. Addition of zinc salts 1 h before or 24 h after inoculation with C. trachomatis was found to inhibit growth. Both C. trachomatis serotype D and a lymphogranuloma venereum strain were inhibited by the zinc salts. Although the mechanism of inhibition is not known, the continued presence of the zinc appeared necessary for maximal effect. A...

Greenberg, S. B.; Harris, D.; Giles, P.; Martin, R. R.; Wallace, R. J.

1985-01-01

272

Antiproliferative effects of metal complexes of new isatin hydrazones against HCT116, MCF7 and HELA tumour cell lines.  

Science.gov (United States)

New hydrazone ligands (HL) derived from 5-substituted isatins and 1-(4-(2-methoxybenzyl)-6-arylpyridazin-3-yl)hydrazines and its complexes with Co(II) and Cu(II) were synthesized. The new hydrazones and their complexes were characterized by means of elemental, spectral analyses and magnetic studies. Primary cytotoxicity evaluation of HL 5a and the new complexes showed that these complexes could act as anticancer agents since they reduced the growth of samples of human tumour cell lines (HCT116((Colon)), MCF7((Breast)) and HELA((Cervix))) to ?18.5 ?g/mL for the new complexes. PMID:21699460

Kandile, Nadia G; Mohamed, Mansoura I; Ismaeel, Hind M

2012-06-01

273

Gamma-radiaton-induced crosslinking of cell-specific chromosomal nonhistone protein-DNA complexes in HeLa chromatin  

International Nuclear Information System (INIS)

Specific antisera were obtained by injecting rabbits or goats with dehistonized HeLa cell chromatin. Gamma irradiation (10 to 100 krad) of the isolated chromatin increased its immunological reactivity with these specific antisera. Conversely, irradiation of the isolated chromosomal nonhistone proteins or DNA followed by reconstitution resulted in a nearly complete loss of immunological reactivity of the reconstituted complexes. Selective protein dissociation experiments indicated that radiation crosslinked the active nonhistone proteins to the DNA. This interpretation was further substantiated by polyacrylamide gel electrophoresis of the crosslinked proteins

1981-01-01

274

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

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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

275

Improved cell-penetrating peptide–PNA conjugates for splicing redirection in HeLa cells and exon skipping in mdx mouse muscle  

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Steric blocking peptide nucleic acid (PNA) oligonucleotides have been used increasingly for redirecting RNA splicing particularly in therapeutic applications such as Duchenne muscular dystrophy (DMD). Covalent attachment of a cell-penetrating peptide helps to improve cell delivery of PNA. We have used a HeLa pLuc705 cell splicing redirection assay to develop a series of PNA internalization peptides (Pip) conjugated to an 18-mer PNA705 model oligonucleotide with higher activity compared to a P...

Ivanova, Gabriela D.; Arzumanov, Andrey; Abes, Rachida; Yin, Haifang; Wood, Matthew J. A.; Lebleu, Bernard; Gait, Michael J.

2008-01-01

276

Effect of doxorubicin on cell survival and micronuclei formation in HeLa cells exposed to different doses of gamma-radiation  

International Nuclear Information System (INIS)

Purpose: The present study was undertaken to obtain an insight into the combined effects of doxorubicin with radiation on the cell survival and micronuclei induction in HeLa cells. Material and Methods: HeLa S3 cells were allowed to grow till they reached plateau phase, inoculated with 10 ?g/ml doxorubicin hydrochloride and then exposed to 0, 0.5, 1, 2 and 3 Gy ?-radiation. Clonogenicity of cell was measured using the colony forming assay, micronuclei formation using the micronucleus assay. Results: The treatment of HeLa cells with doxorubicin (adriamycin) for 2 hours before exposure to different doses of ?-radiation resulted in a significant and dose-dependent decline in the cell survival and cell proliferation when compared to the PBS+irradiation group. Conversely, the frequency of micronuclei increased in a dose-related manner in both the PBS+irradiation and doxorubicin+irradiation groups. The pretreatment of HeLa cells with doxorubicin before irradiation to various doses of ?-rays resulted in a significant elevation in the frequency of micronuclei when compared with the concurrent PBS+irradiation group. The dose-response relationship for both PBS+irradiation and doxorubicin+irradiation groups was linear. The correlation between cell survival and micronuclei induction was also determined for PBS or doxorubicin+irradiation group, where the clonogenicity of cells declined with the increase in micronuclei formation. The correlation between cell survical and micronuclei induction was linear quadratic for both PBS+irradiation and doxorubicin+irradiation groups. Conclusion: From our study it can be concluded that combination treatment with doxorubicin and radiation increased the genotoxic effect of the either treatment given alone. (orig.)

2000-09-01

277

Combination of aloe-emodin with radiation enhances radiation effects and improves differentiation in human cervical cancer cells.  

Science.gov (United States)

The aim of the present study was to investigate the effects of aloe-emodin (AE) on the radiosensitivity and differentiation of HeLa human cervical cancer cells. Cell proliferation was assessed in the HeLa cervical cancer cell line by a methylthiazolyldiphenyl-tetrazolium bromide assay. Radiosensitivity was determined by a colony?forming assay. Flow cytometry was used for analysis of cell cycle distribution and apoptosis. The expression of ?-H2AX and cyclin B was assessed by western blotting. Alkaline phosphatase (ALP) activity was measured by an ALP activity kit. It was demonstrated that AE inhibited the proliferation of HeLa cells in a concentration- and time-dependent manner, induced G2/M and S phase cell cycle arrest and enhanced the radiosensitivity of HeLa cells. The combination of AE and radiation induced apoptosis, upregulated cyclin B and ?-H2AX expression and further improved ALP activity compared with treatment with AE or radiation alone. AE enhanced the radiosensitivity of HeLa human cervical cancer cells in vitro, inhibited the proliferation of HeLa cells, induced G2/M phase cell cycle arrest and, in combination with radiation, induced the apoptosis and improved the differentiation of HeLa cells. PMID:24920336

Luo, Jinghua; Yuan, Yong; Chang, Pengyu; Li, Dawei; Liu, Zhiqiang; Qu, Yaqin

2014-08-01

278

Elevated Ca2+(i) transients induced by trimethyltin chloride in HeLa cells: types and levels of response.  

Science.gov (United States)

Humans are exposed to organotins, like trimethyltin (TMT) chloride via air, water and food, and intoxication might result in severe health complications. Toxic effects of organotin compounds are well documented, but possible mechanisms remain unclear and only little information is available how organometallic species interact with calcium controlling mechanisms. Therefore, the aim of this work was to investigate the effects of TMT on calcium homeostasis in HeLa S3 cells. Dynamic changes of cytosolic calcium (Ca2+(i)) were monitored using laser-scanning microscopy and fluo-4 loaded cells. Application of TMT resulted in sustained as well as in transient elevations of Ca2+(i). The number of reacting cells was directly correlated to the concentration of TMT used: with 500 microM TMT all cells reacted, with 50 microM TMT 80% and with 5 microM 74%. The fast Ca2+(i)-transients (spikes), measured in single cells, occurred even with 0.25 microM TMT and varied in size and duration. The sustained increase of Ca2+(i), measured as the average over all cells, was dose dependent with an approximately 8% increase for 5 microM TMT, approximately 12.3% for 50 microM and approximately 145% for 500 microM TMT. Moreover, this effect was partly reversible. A second application resulted in a similar sustained rise of Ca2+(i) compared to the first application of TMT, there was also no difference when no calcium was added to the external solution (151+/-10% compared to 145+/-15%; 500 microM TMT). This rise of Ca2+(i) was highly reduced (<10% increase) when the internal calcium stores were depleted before TMT (500 microM) was applied. Our data suggest that TMT influences Ca2+(i)-homeostasis of HeLa S3 cells, which might be related to its toxicity in this cell line. PMID:15670872

Florea, Ana-Maria; Dopp, Elke; Büsselberg, Dietrich

2005-03-01

279

Purification and characterization of the glycoprotein hormone. cap alpha. -subunit-like material secreted by HeLa cells  

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The protein secreted by HeLa cells that cross-reacts with antiserum developed against the ..cap alpha..-subunit of human chorionic gonadotropin (hCG) has been purified approximately 30,000-fold from concentrated culture medium by organic solvent fractionation followed by ion exchange, gel filtration, and lectin affinity chromatography. The final preparation had a specific activity (by RIA) of 6.8 x 10/sup 5/ ng of ..cap alpha../mg of protein and appeared homogeneous by electrophoresis on reducing/denaturing polyacrylamide gels (SDS-PAGE). Amino acid analysis indicated that HeLa-..cap alpha.. had a composition very similar to that of the urinary hCG ..cap alpha..-subunit. However, comparison of hCG-..cap alpha.. and HeLa-..cap alpha.. demonstrated that the tumor-associated subunit was not identical with its normal counterpart. The purified tumor protein had an apparent molecular weight greater than that of the urinary ..cap alpha..-subunit when analyzed by SDS-PAGE, and this difference was even greater when a partially purified preparation was examined by an immunoblot technique (Western). Isoelectric focusing of the HeLa and hCG subunits demonstrated that the tumor protein had a lower pI. Immunoprecipitation and electrophoresis of ..cap alpha..-subunit from HeLa cultures labeled with (/sup 3/H)fucose indicated that the tumor subunit was fucosylated, whereas analysis of hCG-..cap alpha.. hydrosylates by HPLC confirmed previous reports that the placental subunit does not contain fucose. The results indicate that, regardless of whether or not a single ..cap alpha..-subunit gene is being expressed in both normal and neoplastic tissues, posttranslational modifications lead to a highly altered subunit in the tumor. The differences observed may be useful in diagnosing neoplastic vs hyperplastic conditions and may lend insight into the mechanism of ectopic hormone production by tumors.

Cox, G.S.; Rimerman, R.A.

1988-08-23

280

Cancer-initiating cells derived from established cervical cell lines exhibit stem-cell markers and increased radioresistance  

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Abstract Background Cancer-initiating cells (CICs) are proposed to be responsible for the generation of metastasis and resistance to therapy. Accumulating evidences indicates CICs are found among different human cancers and cell lines derived from them. Few studies address the characteristics of CICs in cervical cancer. We identify biological features of CICs from four of the best-know human cell lines from uterine cervix tumors. (HeLa, SiHa, Ca Ski, C-4 I). Methods

López Jacqueline; Poitevin Adela; Mendoza-Martínez Veverly; Pérez-Plasencia Carlos; García-Carrancá Alejandro

2012-01-01

 
 
 
 
281

Initiation of poliovirus plus-strand RNA synthesis in a membrane complex of infected HeLa cells  

International Nuclear Information System (INIS)

An in vitro poliovirus RNA-synthesizing system derived from a crude membrance fraction of infected HeLa cells was used to analyze the mechanism of initiation of poliovirus plus-strand RNA synthesis. This system contains an activity that synthesizes the nucleotidyl proteins VPg-pU and VPg-pUpU. These molecules represent the 5'-terminal structure of nascent RNA molecules and of virion RNA. The membranous replication complex is also capable of synthesizing mucleotidyl proteins containing nine or more of the poliovirus 5'-proximal nucleotides as assayed by the formation of the RNase T_1-resistant oligonucleotide VPg-pUUAAAACAGp or by fingerprint analysis of the in vitro-synthesized "3"2P-RNA. Incubation of preformed VPg-pUpU with unlabeled nucleoside triphosphates resulted in the formation of VPg-pUUAAAACAGp. This reaction, which appeared to be an elongation of VPg-pUpU, was stimulated by the addition of a soluble fraction (S-10) obtained from uninfected HeLa cells. Preformed VPg-pU could be chased into VPg-pUpU in the presence of UTP. The data are consistent with a model that VPg-pU can function as a primer for poliovirus plus-strand RNA synthesis in the membranous replication complex and that the elongation reaction may be stimulated by a host cellular factor

1986-01-01

282

Initiation of poliovirus plus-strand RNA synthesis in a membrane complex of infected HeLa cells  

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An in vitro poliovirus RNA-synthesizing system derived from a crude membrance fraction of infected HeLa cells was used to analyze the mechanism of initiation of poliovirus plus-strand RNA synthesis. This system contains an activity that synthesizes the nucleotidyl proteins VPg-pU and VPg-pUpU. These molecules represent the 5'-terminal structure of nascent RNA molecules and of virion RNA. The membranous replication complex is also capable of synthesizing mucleotidyl proteins containing nine or more of the poliovirus 5'-proximal nucleotides as assayed by the formation of the RNase T/sub 1/-resistant oligonucleotide VPg-pUUAAAACAGp or by fingerprint analysis of the in vitro-synthesized /sup 32/P-RNA. Incubation of preformed VPg-pUpU with unlabeled nucleoside triphosphates resulted in the formation of VPg-pUUAAAACAGp. This reaction, which appeared to be an elongation of VPg-pUpU, was stimulated by the addition of a soluble fraction (S-10) obtained from uninfected HeLa cells. Preformed VPg-pU could be chased into VPg-pUpU in the presence of UTP. The data are consistent with a model that VPg-pU can function as a primer for poliovirus plus-strand RNA synthesis in the membranous replication complex and that the elongation reaction may be stimulated by a host cellular factor.

Takeda, N.; Kuhn, R.J.; Yang, C.F.; Takegami, T.; Wimmer, E.

1986-10-01

283

Initiation of poliovirus plus-strand RNA synthesis in a membrane complex of infected HeLa cells.  

Science.gov (United States)

An in vitro poliovirus RNA-synthesizing system derived from a crude membrane fraction of infected HeLa cells was used to analyze the mechanism of initiation of poliovirus plus-strand RNA synthesis. This system contains an activity that synthesizes the nucleotidyl proteins VPg-pU and VPg-pUpU. These molecules represent the 5'-terminal structure of nascent RNA molecules and of virion RNA. The membranous replication complex is also capable of synthesizing nucleotidyl proteins containing nine or more of the poliovirus 5'-proximal nucleotides as assayed by the formation of the RNase T1-resistant oligonucleotide VPg-pUUAAAACAGp or by fingerprint analysis of the in vitro-synthesized RNA. Incubation of preformed VPg-pUpU with unlabeled nucleoside triphosphates resulted in the formation of VPg-pUUAAAACAGp. This reaction, which appeared to be an elongation of VPg-pUpU, was stimulated by the addition of a soluble fraction (S-10) obtained from uninfected HeLa cells. Preformed VPg-pU could be chased into VPg-pUpU in the presence of UTP. Our data are consistent with a model that VPg-pU can function as a primer for poliovirus plus-strand RNA synthesis in the membranous replication complex and that the elongation reaction may be stimulated by a host cellular factor. PMID:3018300

Takeda, N; Kuhn, R J; Yang, C F; Takegami, T; Wimmer, E

1986-10-01

284

The effects of chloroquine and other weak bases on the accumulation and efflux of digoxin and ouabain in HeLa cells.  

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We have studied the effects of the weak bases chloroquine, NH4Cl and amantadine on the handling of certain cardiac glycosides by HeLa cells. When these weak bases are applied acutely to HeLa cells they have only minor effects on the binding of cardiac glycosides to the sodium pumps and on the recovery of pump function following block. When cells are grown in these weak bases there is a variable (10-30%) reduction in pump numbers. This effect is additive to that of chronic treatment with cardi...

1983-01-01

285

Measurement of Protein 53 Diffusion Coefficient in Live HeLa Cells Using Raster Image Correlation Spectroscopy (RICS  

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Full Text Available We have applied Raster Image Correlation Spectroscopy (RICS technique to characterize the dynamics of protein 53 (p53 in living cells before and after the treatment with DNA damaging agents. HeLa cells expressing Green Fluores-cent Protein (GFP tagged p53 were incubated with and without DNA damaging agents, cisplatin or eptoposide, which are widely used as chemotherapeutic drugs. Then, the diffusion coefficient of GFP-p53 was determined by RICS and it was significantly reduced after the drug treatment while that of the one without drug treatment was not. It is suggested that the drugs induced the interaction of p53 with either other proteins or DNA. Together, our results demonstrated that RICS is able to detect the protein dynamics which may be associated with protein-protein or protein-DNA interactions in living cells and it may be useful for the drug screening.

Harinibytaraya Sreenivasappa

2010-10-01

286

Synergistic interactions of saponins and monoterpenes in HeLa cells, Cos7 cells and in erythrocytes.  

Science.gov (United States)

In phytomedicine complex extracts consisting of phenolics, monoterpenes or saponins are traditionally used. It is often impossible to attribute the biological activity of an extract to one or few compounds. As an explanation of the superior activity of extracts, a synergistic effect of combinations of active compounds has been suggested. Since lipophilic monoterpenes or saponins targeting the biomembrane usually accompany polar polyphenols in phytomedical preparations, we decided to investigate their effect as single substances and in combination to gain further insight into potential synergistic effects of herbal medicine. Combinations of the monoterpenes ?-pinene, thymol and menthol with the monodesmosidic saponins digitonin, aescin, glycyrrhizic acid and Quillaja saponin demonstrated strong synergistic activity. The IC(50) of haemolysis was lowered by a factor of 10-100 from 316?g/ml to 2?g/ml for aescin, 157?g/ml to 11?g/ml for Quillaja saponins and 20?g/ml to 3?g/ml for digitonin when combined with thymol. A similar significant synergistic cytotoxicity occurred both in HeLa and Cos7 cells by combining the ?-pinene, thymol and menthol with the saponins. The IC(50) of glycyrrhizic acid was lowered by a factor 100 from around 300?g/ml to around 1-10?g/ml and the IC(50) of aescin, digitonin and Quillaja saponins about the factor 10. Monoterpenes and monodesmosidic saponins have a common target, the biomembrane, which is present in all animal, fungal and bacterial cells. Disturbance of membrane fluidity and permeability is the mode of action. This activity is non-specific which makes it extremely difficult for bacteria and fungi to develop resistance. This explains the overall success of these molecules as defence chemicals in the plant kingdom. The synergistic effect of combinations of saponins with monoterpenes opens a complete new field of possible applications in medicine to overcome resistance in multidrug resistant microbial and human cell. PMID:21968386

Herrmann, Florian; Wink, Michael

2011-10-15

287

A preliminary investigation of modified alginates as a matrix for gene transfection in a HeLa cell model.  

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Previous reports have demonstrated the effectiveness of chitosan as a transfection agent. These studies have noted the importance of polysaccharide backbone interactions with the cell surface as well as cationic groups in the transfection process. The present study focuses upon the potential utility of another polysaccharide hydrogel, alginic acid, as a transfection agent. Alginic acid was modified by carbodiimide-mediated linkage of several heterocyclic and aromatic amines to the carboxyl group of the alginate, giving the alginate polycationic characteristics through which binding to nucleic acids could be facilitated. The amines used for this modification include diaminoacridine, thionin, basic fuchsin, acridine yellow, and diaminomethyltriazine. Of all the conjugates tested, basic fuchsin-modified alginate produced the greatest increase in the transfection of a plasmid coding for beta-galactosidase into HeLa cells. These studies demonstrate that other polysaccharide hydrogels can be used as transfection agents, and the structural orientation of the cationic spacer arm is crucial for effective transfection. PMID:11852701

Padmanabhan, Kiran; Smith, Timothy J

2002-01-01

288

Evidence from uv transcription mapping in HeLa cells that heterogeneous nuclear RNA is the messenger RNA precursor  

International Nuclear Information System (INIS)

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

289

Biological active peptides in human urine: III. Inhibitors of the growth of human leukemia, osteosarcoma, and HeLa cells.  

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Medium-sized peptides isolated from normal humans urine were tested for their effect on DNA, RNA, and protein synthesis, and mitosis, in tissue culture of human myeloblastic leukemia, osteosarcoma, and HeLa cells. Two types of antineoplastic peptides were found. One type consists of strongly acidic peptides (probably sulfated glycopeptides) which act specifically on different kinds of neoplasma. The other type comprises slightly acidic and neutral peptides, and has broad specificity. The active peptides produce up to 97% inhibition of DNA synthesis and mitosis in the neoplastic cells in tissue culture. The peptide fraction which has broad specificity was tested in different concentrations and gave good dose-response relationship. PMID:1066715

Burzynski, S R; Loo, T L; Ho, D H; Rao, P N; Georgiades, G; Kratzenstein, H

1976-01-01

290

Virus-receptor interaction in the adenovirus system I. Identification of virion attachment proteins of the HeLa cell plasma membrane.  

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Plasma membranes from HeLa cells were isolated in a two-phase polymer system. To compare the efficiency of attachment protein extraction, a normalized assay for the assessment of adenovirus type 2 (Ad2) receptor-active components interfering with the attachment of Ad2 to HeLa cells was developed. An optimized detergent extraction procedure, 0.5% Triton X-100, was used, and solubilized membrane proteins were radioisotope labeled in vitro. Proteins with affinity for Ad2 virions were quantified ...

Svensson, U.; Persson, R.; Everitt, E.

1981-01-01

291

The effect of combined treatment of HeLa cells with cisplatin and irradiation upon survival and recovery from radiation damage  

International Nuclear Information System (INIS)

Combined treatment of HeLa cells with cisplatin and irradiation under aerobic conditions leads to no interaction between the two treatment modalities, either when they are given simultaneously or when cisplatin treatment is followed by irradiation. Simultaneous treatment with cisplatin and irradiation under hypoxic conditions leads to radiosensitization of HeLa cells by cisplatin. This sensitization was independent from the surviving fraction with cisplatin only, the dose modifying factor being approximately 1.2. Pretreatment or post-treatment with cisplatin dose not influence the recovery from sublethal radiation damage. (Auth.)

1987-01-01

292

Microinjection of ubiquitin: changes in protein degradation in HeLa cells subjected to heat-shock  

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Ubiquitin was radiolabeled by reaction with 125I-Bolton-Hunter reagent and introduced into HeLa cells using erythrocyte-mediated microinjection. The injected cells were then incubated at 45 degrees C for 5 min (reversible heat-shock) or for 30 min (lethal heat-shock). After either treatment, there were dramatic changes in the levels of ubiquitin conjugates. Under normal culture conditions, approximately 10% of the injected ubiquitin is linked to histones, 40% is found in conjugates with molecular weights greater than 25,000, and the rest is unconjugated. After heat-shock, the free ubiquitin pool and the level of histone-ubiquitin conjugates decreased rapidly, and high molecular weight conjugates predominated. Formation of large conjugates did not require protein synthesis; when analyzed by two-dimensional electrophoresis, the major conjugates did not co-migrate with heat-shock proteins before or after thermal stress. Concomitant with the loss of free ubiquitin, the degradation of endogenous proteins, injected hemoglobin, BSA, and ubiquitin was reduced in heat-shocked HeLa cells. After reversible heat-shock, the decrease in proteolysis was small, and both the rate of proteolysis and the size of the free ubiquitin pool returned to control levels upon incubation at 37 degrees C. In contrast, neither proteolysis nor free ubiquitin pools returned to control levels after lethal heat-shock. However, lethally heat-shocked cells degraded denatured hemoglobin more rapidly than native hemoglobin and ubiquitin-globin conjugates formed within them. Therefore, stabilization of proteins after heat-shock cannot be due to the loss of ubiquitin conjugation or inability to degrade proteins that form conjugates with ubiquitin

1987-01-01

293

Detecção da citotoxicidade de materiais biocompatíveis nas linhagens celulares MRC-5, HeLa e RC-IAL MRC-5, HeLa and RC-IAL cell lines sensitivity for detection of cytotoxicity of biocompatible materials  

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Full Text Available A sensibilidade de uma linhagem celular diplóide e duas heteroplóides, para a detecção de citotoxicidade através do método de difusão em camada de ágar sobre culturas celulares, foi avaliada experimentalmente com solução de ácido ascórbico em diferentes concentrações e, na prática, frente a 562 amostras de 21 diferentes materiais industriais enviados para análise na Seção de Culturas Celulares do Instituto Adolfo Lutz. A linhagem celular heteroplóide designada RC-IAL apresentou, em relação às linhagens MRC-5 e HeLa, maior sensibilidade porque revelou a presença de efeito citotóxico nas menores concentrações utilizadas (10 e 25 ug/ml do ácido ascórbico e apresentou maior diâmetro do halo citotóxico em 15 amostras e igual diâmetro em 16 das 43 amostras (7,6% que resultaram positivas. Nas 43 amostras positivas, a linhagem MRC-5 não revelou citotoxicidade em 3 amostras de espuma e 1 de resina acrílica. O polivinilcloreto (PVC e o polietileno, raramente revelaram positividade, enquanto plástico, algodão e resinas acrílicas revelaram citotoxicidade ao redor de 5%. Em vista dos resultados é discutida a proposta da utilização da linhagem RC-IAL e HeLa para a continuidade das futuras análises solicitadas ao Instituto Adolfo LutzThe 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 25ug/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

Aurea S. Cruz

1992-04-01

294

Detecção da citotoxicidade de materiais biocompatíveis nas linhagens celulares MRC-5, HeLa e RC-IAL / MRC-5, HeLa and RC-IAL cell lines sensitivity for detection of cytotoxicity of biocompatible materials  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: Portuguese Abstract in portuguese A sensibilidade de uma linhagem celular diplóide e duas heteroplóides, para a detecção de citotoxicidade através do método de difusão em camada de ágar sobre culturas celulares, foi avaliada experimentalmente com solução de ácido ascórbico em diferentes concentrações e, na prática, frente a 562 amos [...] tras de 21 diferentes materiais industriais enviados para análise na Seção de Culturas Celulares do Instituto Adolfo Lutz. A linhagem celular heteroplóide designada RC-IAL apresentou, em relação às linhagens MRC-5 e HeLa, maior sensibilidade porque revelou a presença de efeito citotóxico nas menores concentrações utilizadas (10 e 25 ug/ml) do ácido ascórbico e apresentou maior diâmetro do halo citotóxico em 15 amostras e igual diâmetro em 16 das 43 amostras (7,6%) que resultaram positivas. Nas 43 amostras positivas, a linhagem MRC-5 não revelou citotoxicidade em 3 amostras de espuma e 1 de resina acrílica. O polivinilcloreto (PVC) e o polietileno, raramente revelaram positividade, enquanto plástico, algodão e resinas acrílicas revelaram citotoxicidade ao redor de 5%. Em vista dos resultados é discutida a proposta da utilização da linhagem RC-IAL e HeLa para a continuidade das futuras análises solicitadas ao Instituto Adolfo Lutz Abstract in english 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, show [...] ed 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 25ug/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, Aurea S.; Figueiredo, Cristina A.; Martinez, Clélia H. O.; Salles Gomes, Luís F. de.

295

Effects of metal treatment on DNA repair in polyamine-depleted HeLa cells with special reference to nickel.  

Science.gov (United States)

Human cells depleted of the naturally occurring polyamines putrescine, spermidine, and spermine exhibit altered chromatin structure and marked deficiencies in DNA replicative and repair processes. Similar effects have been observed following treatment of normal mammalian cells with various heavy metal salts. In an attempt to better understand how metals interfere with normal DNA metabolic processes, a series of studies was carried out in which the toxicity and repair-inhibitory properties of various metals were evaluated in polyamine-depleted HeLa cells. Cytotoxicity of copper, zinc, magnesium, and cadmium was not altered in cells carrying lower polyamine pools. However, the sensitivity to nickel was markedly increased upon polyamine depletion, a condition that was readily reversed by polyamine supplementation. Nucleoid sedimentation analysis indicated that a greater amount of nickel-induced DNA damage occurred in polyamine-depleted cells than in normal cells, possibly serving as the basis for the increased sensitivity. Both polyamine depletion and nickel treatment result in decreased repair of DNA strand breaks and decreased cloning efficiency following X-ray and ultraviolet irradiation. Nickel treatment of polyamine-depleted cells resulted in synergistic sensitivity to both radiation treatments. None of the other metals tested enhanced X-ray or ultraviolet sensitivity of polyamine-depleted cells. Analysis of retarded repair sites following ultraviolet irradiation indicated those sites to be nonligatable in polyamine-depleted and nickel-treated cells, suggesting a block in the normal gap-sealing process. PMID:7843137

Snyder, R D

1994-09-01

296

Functional analysis of a recombinant PIII-SVMP, GST-acocostatin; an apoptotic inducer of HUVEC and HeLa, but not SK-Mel-28 cells  

Science.gov (United States)

Disintegrins and disintegrin-like peptides interact with integrins and interfere with cell-cell and cell-matrix interactions. A disintegrin-like snake venom gene, Acocostatin was cloned from the venom gland mRNA of Agkistrodon contortrix contortrix. Acocostatin belongs to the PIII-SVMP subfamily of disintegrin-like peptides. The recombinant acocostatin peptide was produced and purified as GST-fusion. The GST-acocostatin peptide, at 44 ?g/mL, inhibited platelet aggregation by 30% in PRP and 18% in whole blood. In addition GST-acocostatin, at 220?g/mL, inhibited SK-Mel-28 cell migration by 48%, but did not inhibit T24 cell migration. The GST-acocostatin peptide ability to induce apoptosis on HUVEC, HeLa, and SK-Mel-28 cells was determined using Annexin-V-FITC and chromatin fragmentation assays after 24 h of treatment. At 5 ?M GST-acocostatin peptide, 19.68% +/? 3.09 of treated HUVEC, and 35.86% +/? 2.05 of treated HeLa cells were in early apoptosis. The GST-acocostatin peptide also caused chromatin fragmentation of HUVEC and HeLa cells as determined by fluorescent microscopy and Hoechst staining. The GST-acocostatin peptide failed to induce apoptosis of SK-Mel-28 cells. We characterized the HUVEC, HeLa, and T24 integrin expression by flow cytometry, as the first step in determining GST-acocostatin binding specificity. Our results indicate that HUVEC express ?v, ?v?3, ?v?5, ?6, ?1, and ?3 integrin receptors. HeLa cells express ?1, ?2, ?6, ?v, ?v?5, and ?1 integrin receptors. T24 cells express ?1, ?3, ?6, ?v, ?v?3, ?v?5, ?1, ?3, and ?6 integrin receptors.

Teklemariam, Takele; Seoane, Agustin I.; Ramos, Carla J.; Sanchez, Elda E.; Lucena, Sara E.; Perez, John C.; Mandal, Stephanie A.; Soto, Julio G.

2011-01-01

297

An in-cell NMR study of monitoring stress-induced increase of cytosolic Ca2+ concentration in HeLa cells  

International Nuclear Information System (INIS)

Highlights: •We performed time-resolved NMR observations of calbindin D9k in HeLa cells. •Stress-induced increase of cytosolic Ca2+ concentration was observed by in-cell NMR. •Calbindin D9k showed the state-transition from Mg2+- to Ca2+-bound state in cells. •We provide a useful tool for in situ monitoring of the healthiness of the cells. -- Abstract: Recent developments in in-cell NMR techniques have allowed us to study proteins in detail inside living eukaryotic cells. The lifetime of in-cell NMR samples is however much shorter than that in culture media, presumably because of various stresses as well as the nutrient depletion in the anaerobic environment within the NMR tube. It is well known that Ca2+-bursts occur in HeLa cells under various stresses, hence the cytosolic Ca2+ concentration can be regarded as a good indicator of the healthiness of cells in NMR tubes. In this study, aiming at monitoring the states of proteins resulting from the change of cytosolic Ca2+ concentration during experiments, human calbindin D9k (P47M + C80) was used as the model protein and cultured HeLa cells as host cells. Time-resolved measurements of 2D 1H–15N SOFAST–HMQC experiments of calbindin D9k (P47M + C80) in HeLa cells showed time-dependent changes in the cross-peak patterns in the spectra. Comparison with in vitro assignments revealed that calbindin D9k (P47M + C80) is initially in the Mg2+-bound state, and then gradually converted to the Ca2+-bound state. This conversion process initiates after NMR sample preparation. These results showed, for the first time, that cells inside the NMR tube were stressed, presumably because of cell precipitation, the lack of oxygen and nutrients, etc., thereby releasing Ca2+ into cytosol during the measurements. The results demonstrated that in-cell NMR can monitor the state transitions of stimulated cells through the observation of proteins involved in the intracellular signalling systems. Our method provides a very useful tool for in situ monitoring of the “healthiness” of the cells in various in-cell NMR studies

2013-09-06

298

An in-cell NMR study of monitoring stress-induced increase of cytosolic Ca{sup 2+} concentration in HeLa cells  

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Highlights: •We performed time-resolved NMR observations of calbindin D{sub 9k} in HeLa cells. •Stress-induced increase of cytosolic Ca{sup 2+} concentration was observed by in-cell NMR. •Calbindin D{sub 9k} showed the state-transition from Mg{sup 2+}- to Ca{sup 2+}-bound state in cells. •We provide a useful tool for in situ monitoring of the healthiness of the cells. -- Abstract: Recent developments in in-cell NMR techniques have allowed us to study proteins in detail inside living eukaryotic cells. The lifetime of in-cell NMR samples is however much shorter than that in culture media, presumably because of various stresses as well as the nutrient depletion in the anaerobic environment within the NMR tube. It is well known that Ca{sup 2+}-bursts occur in HeLa cells under various stresses, hence the cytosolic Ca{sup 2+} concentration can be regarded as a good indicator of the healthiness of cells in NMR tubes. In this study, aiming at monitoring the states of proteins resulting from the change of cytosolic Ca{sup 2+} concentration during experiments, human calbindin D{sub 9k} (P47M + C80) was used as the model protein and cultured HeLa cells as host cells. Time-resolved measurements of 2D {sup 1}H–{sup 15}N SOFAST–HMQC experiments of calbindin D{sub 9k} (P47M + C80) in HeLa cells showed time-dependent changes in the cross-peak patterns in the spectra. Comparison with in vitro assignments revealed that calbindin D{sub 9k} (P47M + C80) is initially in the Mg{sup 2+}-bound state, and then gradually converted to the Ca{sup 2+}-bound state. This conversion process initiates after NMR sample preparation. These results showed, for the first time, that cells inside the NMR tube were stressed, presumably because of cell precipitation, the lack of oxygen and nutrients, etc., thereby releasing Ca{sup 2+} into cytosol during the measurements. The results demonstrated that in-cell NMR can monitor the state transitions of stimulated cells through the observation of proteins involved in the intracellular signalling systems. Our method provides a very useful tool for in situ monitoring of the “healthiness” of the cells in various in-cell NMR studies.

Hembram, Dambarudhar Shiba Sankar; Haremaki, Takahiro; Hamatsu, Jumpei; Inoue, Jin; Kamoshida, Hajime; Ikeya, Teppei; Mishima, Masaki [Department of Chemistry, Graduate School of Science and Engineering, Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji-shi, Tokyo 192-0373 (Japan); Mikawa, Tsutomu [Cellular and Molecular Biology Unit, RIKEN Advanced Science Institute, Wako-shi, Saitama 351-0198 (Japan); Hayashi, Nobuhiro [Department of Life Science, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 B-1, Nagatsuda-chou, Midori-ku, Yokohama, Kanagawa 226-8501 (Japan); Shirakawa, Masahiro [Department of Molecular Engineering, Graduate School of Engineering, Kyoto University, Nishikyo-ku, Kyoto 615-8510 (Japan); Ito, Yutaka, E-mail: ito-yutaka@tmu.ac.jp [Department of Chemistry, Graduate School of Science and Engineering, Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji-shi, Tokyo 192-0373 (Japan)

2013-09-06

299

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 conjugated with glutathione to form thiodione. Thiodione is then recognized and transported across the cell membrane via the ATP-driven MRP1 pump. In the extracellular environment, thiodione was detected by...

2012-01-01

300

Microarray Analysis Reveals Characteristic Changes of Host Cell Gene Expression in Response to Attenuated Modified Vaccinia Virus Ankara Infection of Human HeLa Cells  

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The potential use of the modified vaccinia virus Ankara (MVA) strain as a live recombinant vector to deliver antigens and elicit protective immune responses against infectious diseases demands a comprehensive understanding of the effect of MVA infection on human host gene expression. We used microarrays containing more than 15,000 human cDNAs to identify gene expression changes in human HeLa cell cultures at 2, 6, and 16 h postinfection. Clustering of the 410 differentially regulated genes id...

Guerra, Susana; Lo?pez-ferna?ndez, Luis A.; Conde, Raquel; Pascual-montano, Alberto; Harshman, Keith; Esteban, Mariano

2004-01-01

 
 
 
 
301

Ethanolic extract of the Goldenseal, Hydrastis canadensis, has demonstrable chemopreventive effects on HeLa cells in vitro: Drug-DNA interaction with calf thymus DNA as target.  

Science.gov (United States)

This study tested chemotherapeutic potential of Hydrastis canadensis (HC) extract in HeLa cells in vitro, with emphasis on its drug-DNA interaction and apoptosis induction ability. Nuclear uptake of HC by DAPI, Ao/Eb staining and internucleosomal DNA damage by comet assay was studied through fluorescence microscopy. Possible changes in MMP and apoptotic signalling events were critically analyzed. Cell cycle progression studied through FACS and fragmented DNA through "TUNEL" assay were critically analyzed. RT-PCR studies were conducted for analyzing Cyt-C and Bax translocation in mitochondrial and cytosolic extracts, and Caspase 3 in whole cell lysate. Role of p53-mediated regulation of NF-?? and TNF-? was elucidated by Western blot analysis. Data of CD and Tm profile of CT-DNA were analyzed. Overall results indicated anti-cancer potential of HC through its ability to induce apoptosis, and interaction with CT-DNA that changed structural conformation of DNA, proving HC to be a promising candidate for chemoprevention. PMID:23628949

Saha, Santu Kumar; Sikdar, Sourav; Mukherjee, Avinaba; Bhadra, Kakali; Boujedaini, Naoual; Khuda-Bukhsh, Anisur Rahman

2013-07-01

302

Effectiveness of combined treatment using X-rays and a phosphoinositide 3-kinase inhibitor, ZSTK474, on proliferation of HeLa cells in vitro and in vivo  

International Nuclear Information System (INIS)

ZSTK474 is a novel orally applicable phosphoinositide 3-kinase-specific inhibitor that strongly inhibits cancer cell proliferation. To further explore the antitumor effect of ZSTK474 for future clinical usage, we studied its combined effects with radiation. The proliferation of HeLa cells was inhibited by treatment with X-rays alone or ZSTK474 alone. Combination treatment using X-rays then ZSTK474 given orally for 8 days, starting 24 h post-irradiation, significantly enhanced cell growth inhibition. The combined effect was also observed for clonogenic survival with continuous ZSTK474 treatment. Western blot analysis showed enhanced phosphorylation of Akt and GSK-3? by X-irradiation, whereas phosphorylation was inhibited by ZSTK474 treatment alone. Treatment with ZSTK474 after X-irradiation also inhibited phosphorylation, and remarkably inhibited xenograft tumor growth. Combined treatment with X-rays and ZSTK474 has greater therapeutic potential than radiation or drug therapy alone, both in vitro and in vivo. (author)

2011-06-01

303

Anticancer-cytotoxic activity of saponins isolated from the leaves of Gymnema sylvestre and Eclipta prostrata on HeLa cells  

Directory of Open Access Journals (Sweden)

Full Text Available The anticancer-cytotoxic activities of isolated saponins, gymnemagenol (C 30 H 50 O 4 from Gymnema sylvestre and dasyscyphin C (C 28 H 40 O 8 from Eclipta prostrata leaves were tested under in vitro conditions in HeLa cells. The gymnemagenol and dayscyphin C at 50 ?g/ml showed a good cytotoxic activity (63% and 52%, respectively in HeLa cells at 48 hours with the IC50 value of 37 and 50 ?g/ml, respectively. 5-Fluorouracil (5-FU, a positive control, showed 57.5 % cell death with the IC50 value of 36 ?g/ml. The percentage of HeLa cell death was maximum (73% after 96 hours with gymnemagenol, whereas dasyscyphin C showed only 53%. The isolated saponins were not toxic to Vero cells. From this study, it can be concluded that the saponins, gymnemagenol, and dayscyphin C have significant anticancer-cytotoxic activity on HeLa cells under in vitro conditions.

Khanna Venkatesan

2009-01-01

304

Differences in the response of early and mid or late G1 WI38 cells treated with cyclohexamide to HeLa inducers of DNA synthesis  

International Nuclear Information System (INIS)

The inducibility of DNA synthesis after treatment with cyclohexamide (CHM) during mitosis and the G1 phase of WI38 cells has been studied in the heterokaryons following fusion with HeLa cells in S phase. Synchronized mitotic cells treated for up to 5 h with CHM were not delayed in the initiation of DNA synthesis in the heterokaryons. The G1 cells treated with CHM for 3-24 h were slow in responding to inducers of DNA synthesis generated by HeLa cells in the heterokaryons. The results suggest that there is a specific point in early G1 that regulates the entry of cells into a cycling state. In the presence of CHM, mitotic cells divide, but the daughter cells fail to enter G1 leading to DNA synthesis, and CHM treatment of G1 cells results in their transient entry into a G0 state

1987-01-01

305

In Vitro Ultramorphological Assessment of Apoptosis Induced by Zerumbone on (HeLa  

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Full Text Available Zerumbone (ZER, a potential anticancer compound, isolated from the fresh rhizomes of Zingiber zerumbet. In this investigation, the cytotoxic properties of ZER were evaluated, on cancer cells of human cervix (HeLa, breast and ovary, and normal cells of Chinese Hamster ovary, using MTT assay. Apoptogenic effects of ZER on HeLa were studied using fluorescence microscopy (AO/PI double staining, scanning and transmission electron microscopy (SEM and TEM, and colorimetric assay of the apoptosis promoter enzyme, caspase-3. The results of MTT assay showed that ZER has less effect on normal cells compared to cancer cells. The lowest IC50 of ZER was observed on HeLa cells. Cytological observations showed nuclear and chromatin condensation, cell shrinkage, multinucleation, abnormalities of mitochondrial cristae, membrane blebbing, holes, cytoplasmic extrusions and formation of apoptotic bodies as confirmed collectively by double staining of AO/PI, SEM and TEM. Statistical analysis (two-tailed t-test of differential counting of 200 cells under fluorescence microscope revealed significant difference in apoptotic cells populations between treated and untreated HeLa cells. In addition, ZER has increased the cellular level of caspase-3 on the treated HeLa cells. It could be concluded that ZER was able to produce distinctive morphological features of cell death that corresponds to apoptosis.

2009-03-01

306

Fold-back tetraplex DNA species in DNase I-resistant DNA isolated from HeLa cells.  

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A DNase I-resistant DNA species has been isolated and purified from HeLa cells by gel electrophoresis. Our studies indicate that the DNase I-resistant DNA species was about 40-60 bp fragment sizes responding to double-strand DNA marker and has higher guanine content. The image of AFM showed that this species has been assumed to be tetraplex structure according to its apparent width and height. Its CD, UV spectrum also exhibited characteristics similar to some tetraplex structure, which was different from the standard duplex DNA. 32P-labeled probes (TTAGGG)4 and 5'-TGGGGAGGGTGGGGAGGGTGGGGAAGG-3' could be hybridized to purified DNase I-resistant species. All results suggest that the DNase I-resistant DNA species have at least two components, which adopt an intrastrand fold-back DNA tetraplex. Their sequences were similar to human telomere and human c-myc locus (NHE), respectively. PMID:10798531

Cao, E; Sun, X; Zhang, X; Li, J; Bai, C

2000-04-01

307

Formation of products of the 5,6-dihydroxydihydrothymine type by ultraviolet light in HeLa cells  

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The formation of products of the 5,6-dihydroxydihydrothymine-type (t/sup UV/) and cyclobutane-type pyrimidine photodimers (TT) and tritiated water (3H2O) by monochromatic light at 240, 265, 280, and 313 nm (6-nm half-band-width) was investigated in HeLa S-3 cells which were labeled in their DNA with [methyl-3H]thymine. The efficiency of the formation of all three products was maximal at 280 nm and dropped towards longer and shorter wavelengths. The efficiency of TT formation dropped more strongly towards longer and shorter wavelengths than the efficiency of t/sup UV/ formation (comparison at a dose of 5 x 103 J m-2). Total monomeric thymine ring saturation (t/sub sat/) was estimated from the t/sup UV/ data. It was calculated that 0.06 t/sub sat/ was formed for each TT at 280 nm, but 0.73 t/sub sat/ per TT at 313 nm. It follows that monomeric thymine ring saturation represents a minor photochemical reaction relative to pyrimidine dimerization in the far-ultraviolet but a major reaction in the near-ultraviolet. The formation of 3H2O by ultraviolet light from [methyl-3H] thymidine-labeled HeLa cells most likely indicates the attack of hydroxyl radicals on the cellular DNA; 5-methyleneuracil radicals formed as a consequence of the reaction may be important intermediates in ultraviolet-induced DNA-DNA and DNA-protein cross-linking

1977-06-14

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The mTOR inhibitor AZD8055 inhibits proliferation and glycolysis in cervical cancer cells  

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The aim of the present study was to determine the effect of AZD8055 on proliferation, apoptosis and glycolysis in the human cervical cancer cell line HeLa and to investigate the underlying mechanism(s) of action. HeLa human cervical cancer cells were treated with 10 nM AZD8055 for 24, 48 or 72 h. MTT was used to determine cell proliferation. Annexin V/propidium iodide staining was used to determine cell apoptosis analyzed by fluorescence-activated cell sorting (FACS). Glycolytic activity was ...

Li, Shaoru; Li, Yan; Hu, Ruili; Li, Weihua; Qiu, Haifeng; Cai, Honghua; Wang, Shijin

2013-01-01

309

Femtosecond laser nanosurgery of sub-cellular structures in HeLa cells by employing Third Harmonic Generation imaging modality as diagnostic tool.  

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Femtosecond laser assisted nanosurgery of microscopic biological specimens is a relatively new technique which allows the selective disruption of sub-cellular structures without causing any undesirable damage to the surrounding regions. The targeted structures have to be stained in order to be clearly visualized for the nanosurgery procedure. However, the validation of the final nanosurgery result is difficult, since the targeted structure could be simply photobleached rather than selectively destroyed. This fact comprises a main drawback of this technique. In our study we employed a multimodal system which integrates non-linear imaging modalities with nanosurgery capabilities, for the selective disruption of sub-cellular structures in HeLa cancer cells. Third Harmonic Generation (THG) imaging modality was used as a tool for the identification of structures that were subjected to nanosurgery experiments. No staining of the biological samples was required, since THG is an intrinsic property of matter. Furthermore, cells' viability after nanosurgery processing was verified via Two Photon Excitation Fluorescence (TPEF) measurements. PMID:22259045

Tserevelakis, George J; Psycharakis, Stylianos; Resan, Bojan; Brunner, Felix; Gavgiotaki, Evagelia; Weingarten, Kurt; Filippidis, George

2012-02-01

310

The effect of photodynamic treatment on the morphological and mechanical properties of the HeLa cell line.  

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High resolution imaging of biological structures and changes induced by various agents such as drugs and toxins is commonly performed by fluorescence and electron microscopy (EM). Although high-resolution imaging is possible with EM, the requirements for fixation and staining of samples for image contrast severely limits the study of living organisms. Atomic force microscopy (AFM), on the other hand, is capable of simultaneous nanometer spatial resolution and piconewton force detection, allowing detailed study of cell surface morphology and monitoring cytomechanical information. We present a method that images and studies mechanically characterized cells using AFM. We used a HeLa cell line (cervix carcinoma cell), which is sensitive to photodynamic treatment (PDT); growth media as a scanning surrounding; atomic force microscopy NT-MDT Aura for cytomechanical measurement; and scanning electron microscope Hitachi Su 6600 for control images of the cells. The modulus of elasticity for intact and photodynamically damaged cells can indicate mechanical changes to the main properties of cells. Cell elasticity changes can provide information on the degree or value of cell damage, for example after PDT. Measurements were carried out on approximately sixty cells, including three independent experiments on a control group and on sixty cells in a photodamaged group. Cells before PDT show higher elasticity: the median of Young´s modulus on the nucleus was 35.283 kPa and outside of the nucleus 107.442 kPa. After PDT, the median of Young's modulus on the nucleus was 61.144 kPa and outside of the nucleus was 193.605 kPa. PMID:23817636

Kolar, Petr; Tomankova, Katerina; Malohlava, Jakub; Zapletalova, Jana; Vujtek, Milan; Safarova, Klara; Jancik, Dalibor; Kolarova, Hana

2013-09-01

311

FOXL2 suppresses proliferation, invasion and promotes apoptosis of cervical cancer cells.  

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FOXL2 is a transcription factor that is essential for ovarian function and maintenance, the germline mutations of which give rise to the blepharophimosis ptosis epicanthus inversus syndrome (BPES), often associated with premature ovarian failure. Recently, its mutations have been found in ovarian granulosa cell tumors (OGCTs). In this study, we measured the expression of FOXL2 in cervical cancer by immunohistochemistry and its mRNA level in cervical cancer cell lines Hela and Siha by RT-PCR. Then we overexpressed FOXL2 in Hela cells and silenced it in Siha cells by plasmid transfection and verified using western blotting. When FOXL2 was overexpressed or silenced, cells proliferation and apoptosis were determined by Brdu assay and Annexin V/PI detection kit, respectively. In addition, we investigated the effects of FOXL2 on the adhesion and invasion of Hela and Siha cells. Finally, we analyzed the influences of FOXL2 on Ki67, PCNA and FasL by flow cytometry. The results showed that FOXL2 was highly expressed in cervical squamous cancer. Overexpressing FOXL2 suppressed Hela proliferation and facilitated its apoptosis. Silencing FOXL2 enhanced Siha proliferation and inhibited its apoptosis. Meanwhile, silencing FOXL2 promoted Siha invasion, but it had no effect on cells adhesion. In addition, overexpressing FOXL2 decreased the expression of Ki67 in Hela and Siha cells. Therefore, our results suggested that FOXL2 restrained cells proliferation and enhanced cells apoptosis mainly through decreasing Ki67 expression. PMID:24817949

Liu, Xing-Long; Meng, Yu-Han; Wang, Jian-Li; Yang, Biao-Bing; Zhang, Fan; Tang, Sheng-Jian

2014-01-01

312

FOXL2 suppresses proliferation, invasion and promotes apoptosis of cervical cancer cells  

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FOXL2 is a transcription factor that is essential for ovarian function and maintenance, the germline mutations of which give rise to the blepharophimosis ptosis epicanthus inversus syndrome (BPES), often associated with premature ovarian failure. Recently, its mutations have been found in ovarian granulosa cell tumors (OGCTs). In this study, we measured the expression of FOXL2 in cervical cancer by immunohistochemistry and its mRNA level in cervical cancer cell lines Hela and Siha by RT-PCR. Then we overexpressed FOXL2 in Hela cells and silenced it in Siha cells by plasmid transfection and verified using western blotting. When FOXL2 was overexpressed or silenced, cells proliferation and apoptosis were determined by Brdu assay and Annexin V/PI detection kit, respectively. In addition, we investigated the effects of FOXL2 on the adhesion and invasion of Hela and Siha cells. Finally, we analyzed the influences of FOXL2 on Ki67, PCNA and FasL by flow cytometry. The results showed that FOXL2 was highly expressed in cervical squamous cancer. Overexpressing FOXL2 suppressed Hela proliferation and facilitated its apoptosis. Silencing FOXL2 enhanced Siha proliferation and inhibited its apoptosis. Meanwhile, silencing FOXL2 promoted Siha invasion, but it had no effect on cells adhesion. In addition, overexpressing FOXL2 decreased the expression of Ki67 in Hela and Siha cells. Therefore, our results suggested that FOXL2 restrained cells proliferation and enhanced cells apoptosis mainly through decreasing Ki67 expression.

Liu, Xing-Long; Meng, Yu-Han; Wang, Jian-Li; Yang, Biao-Bing; Zhang, Fan; Tang, Sheng-Jian

2014-01-01

313

Increased expression of cyclin B1 mRNA coincides with diminished G2-phase arrest in irradiated HeLa cells treated with staurosporine or caffeine  

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The irradiation of cells results in delayed progression through the G2 phase of the cell cycle. Treatment of irradiated HeLa cells with caffeine greatly reduces the G2-phase delay, while caffeine does not alter progression of cells through the cell cycle in unirradiated cells. In this report we demonstrate that treatment of HeLa cells with the kinase inhibitor staurosporine, but not with the inhibitor H7, also results in a reduction of the G2-phase arrest after irradiation. Cell cycle progression in unirradiated cells is unaffected by 4.4 nM (2ng/ml) staurosporine, which releases the radiation-induced G2-phase arrest. In HeLa cells, the G2-phase delay after irradiation in S phase is accompanied by decreased expression of cyclin B1 mRNA. Coincident with the reduction in G2-phase delay, we observed an increase in cyclin B1 mRNA accumulation in irradiated, staurosporine-treated cells compared to cells treated with irradiation alone. Caffeine treatment of irradiated HeLa cells also resulted in an elevation in the levels of cyclin B1 message. These results support the hypothesis that diminished cyclin B1 mRNA levels influence G2-phase arrest to some degree. The findings that both staurosporine and caffeine treatments reverse the depression in cyclin B1 expression suggest that these two compounds may act on a common pathway of cell cycle control in response to radiation injury. 33 refs., 6 figs

1994-12-01

314

RGDS-functionalized polyethylene glycol hydrogel-coated magnetic iron oxide nanoparticles enhance specific intracellular uptake by HeLa cells  

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Full Text Available Caner Nazli1, Tugba Ipek Ergenc2, Yasemin Yar1, Havva Yagci Acar1,3, Seda Kizilel1,21Graduate School of Sciences and Engineering, Koç University, 2Department of Chemical and Biological Engineering, College of Engineering, Koç University, 3Department of Chemistry, Faculty of Arts and Sciences, Koç University, Istanbul, TurkeyAbstract: The objective of this study was to develop thin, biocompatible, and biofunctional hydrogel-coated small-sized nanoparticles that exhibit favorable stability, viability, and specific cellular uptake. This article reports the coating of magnetic iron oxide nanoparticles (MIONPs with covalently cross-linked biofunctional polyethylene glycol (PEG hydrogel. Silanized MIONPs were derivatized with eosin Y, and the covalently cross-linked biofunctional PEG hydrogel coating was achieved via surface-initiated photopolymerization of PEG diacrylate in aqueous solution. The thickness of the PEG hydrogel coating, between 23 and 126 nm, was tuned with laser exposure time. PEG hydrogel-coated MIONPs were further functionalized with the fibronectin-derived arginine-glycine-aspartic acid-serine (RGDS sequence, in order to achieve a biofunctional PEG hydrogel layer around the nanoparticles. RGDS-bound PEG hydrogel-coated MIONPs showed a 17-fold higher uptake by the human cervical cancer HeLa cell line than that of amine-coated MIONPs. This novel method allows for the coating of MIONPs with nano-thin biofunctional hydrogel layers that may prevent undesirable cell and protein adhesion and may allow for cellular uptake in target tissues in a specific manner. These findings indicate that the further biofunctional PEG hydrogel coating of MIONPs is a promising platform for enhanced specific cell targeting in biomedical imaging and cancer therapy.Keywords: PEG hydrogel, surface-initiated photopolymerization, nanoparticle encapsulation, agglomeration

Nazli C

2012-04-01

315

Modulation of intracellular calcium homeostasis by trimethyltin chloride in human tumour cells: Neuroblastoma SY5Y and cervix adenocarcinoma HeLa S3  

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Physiological modifications of intracellular Ca2+ ([Ca2+]i) levels trigger and/or regulate a diversity of cellular activities (e.g. neurotransmitter release, synaptic plasticity, muscular contraction, cell proliferation), while calcium overloads could result in cytotoxicity. Previously, we have shown that trimethyltin chloride (Me3SnCl; TMT) modulates calcium homeostasis in cervix adenocarcinoma (HeLa S3) cells [Florea, A.-M., Dopp, E., Buesselberg, D., 2005. TMT induces elevated calcium transients in HeLa cells: types and levels of response. Cell Calcium 37, 252-258]. Here we compare [Ca2+]i-changes induced by trimethyltin chloride in neuroblastoma SY5Y and HeLa S3 cells using calcium-sensitive dyes (fluo-4/AM (fluo-4) and rhod-2/AM (rhod-2)) and laser scanning microscopy (LSM). TMT-induced calcium elevations in neuroblastoma SY5Y as well as in HeLa S3 cells. [Ca2+]i rose to a sustained plateau or to transient spikes. Overall, the detected averaged increase of the maximum calcium elevation were: 0.5 ?M ?125.6%; 5 ?M ?130.1%; 500 ?M ?145% in HeLa S3 cells and 0.5 ?M ?133.3%; 5 ?M ?136.1%; 500 ?M ?147.1% in neuroblastoma SY5Y cells. The calcium rise derived from internal stores did not significantly depend on the presence of calcium in the external solution: ?109% (no calcium added) versus ?117% (2 mM calcium; 5 ?M TMT) in HeLa cells. This difference was similar in neuroblastoma SY5Y cells, were ?127% versus ?136% increase (5 ?M TMT) were measured. Staining of calcium stores with rhod-2 showed a TMT-induced [Ca2+]i-decrease in the stores followed by an increase of the calcium concentration in the nuclei of the two cell lines tested. Our results suggest that toxic effects in human tumour cells after exposure to trimethyltin compounds might be due to an elevation of [Ca2+]i

2005-12-01

316

Modulation of intracellular calcium homeostasis by trimethyltin chloride in human tumour cells: neuroblastoma SY5Y and cervix adenocarcinoma HeLa S3.  

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Physiological modifications of intracellular Ca2+ ([Ca2+]i) levels trigger and/or regulate a diversity of cellular activities (e.g. neurotransmitter release, synaptic plasticity, muscular contraction, cell proliferation), while calcium overloads could result in cytotoxicity. Previously, we have shown that trimethyltin chloride (Me3SnCl; TMT) modulates calcium homeostasis in cervix adenocarcinoma (HeLa S3) cells [Florea, A.-M., Dopp, E., Büsselberg, D., 2005. TMT induces elevated calcium transients in HeLa cells: types and levels of response. Cell Calcium 37, 252-258]. Here we compare [Ca2+]i-changes induced by trimethyltin chloride in neuroblastoma SY5Y and HeLa S3 cells using calcium-sensitive dyes (fluo-4/AM (fluo-4) and rhod-2/AM (rhod-2)) and laser scanning microscopy (LSM). TMT-induced calcium elevations in neuroblastoma SY5Y as well as in HeLa S3 cells. [Ca2+]i rose to a sustained plateau or to transient spikes. Overall, the detected averaged increase of the maximum calcium elevation were: 0.5 microM approximately 125.6%; 5 microM approximately 130.1%; 500 microM approximately 145% in HeLa S3 cells and 0.5 microM approximately 133.3%; 5 microM approximately 136.1%; 500 microM approximately 147.1% in neuroblastoma SY5Y cells. The calcium rise derived from internal stores did not significantly depend on the presence of calcium in the external solution: approximately 109% (no calcium added) versus approximately 117% (2 mM calcium; 5 microM TMT) in HeLa cells. This difference was similar in neuroblastoma SY5Y cells, were approximately 127% versus approximately 136% increase (5 microM TMT) were measured. Staining of calcium stores with rhod-2 showed a TMT-induced [Ca2+]i-decrease in the stores followed by an increase of the calcium concentration in the nuclei of the two cell lines tested. Our results suggest that toxic effects in human tumour cells after exposure to trimethyltin compounds might be due to an elevation of [Ca2+]i. PMID:16125831

Florea, Ana-Maria; Splettstoesser, Frank; Dopp, Elke; Rettenmeier, Albert W; Büsselberg, Dietrich

2005-12-01

317

Phototoxic effect of TPPS4 and MgTPPS4 on DNA fragmentation of HeLa cells.  

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Photodynamic therapy (PDT) is an alternative method of tumour treatment. It is based on a photochemical reaction of a photosensitizer, irradiation, and O(2) which converts to cytotoxic (1)O(2) and other forms of reactive oxygen species (ROS). The comet assay (also called single-cell gel electrophoresis, SCGE) is a sensitive, simple and quantitative technique for detection of DNA damage. In our study we investigated the phototoxicity of the two porphyrin photosensitizers, TPPS4 and MgTPPS4, on HeLa cells. Three different radiation doses and six different concentrations of the photosensitizers were used. Our results show that the DNA of the cells treated with the TPPS(4) and MgTPPS(4) at the concentrations higher than 5 ?M was highly fragmented indicating a strong phototoxic effect resulting in a cell apoptosis. On the base of our results we can hypothesize that even the irradiation dose of 1 J cm(-2) is sufficient enough to provoke the DNA fragmentation. PMID:21078379

Binder, S; Kolarova, H; Tomankova, K; Bajgar, R; Daskova, A; Mosinger, J

2011-09-01

318

Some transplantable substance in spent medium obtained from the plateau culture of HeLa cells affecting the sensitivity to bleomycin  

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The cytotoxic effect of bleomycin (BLM) on HeLa cells grown at plateau phase was investigated and compared with that at exponential phase. Cells were cultured in F10 medium supplemented 10% calf serum and antibiotics before exposure to BLM. Results show that the sensitivity seen at low dose and short time exposures was extended in the spent medium. It is suggested that there is some unknown substance in the spent medium that enhances cell killing by bleomycin.

Miyamoto, T.; Terasima, T.

1986-05-01

319

Queuosine Biosynthesis Is Required for Sinorhizobium meliloti-Induced Cytoskeletal Modifications on HeLa Cells and Symbiosis with Medicago truncatula  

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Rhizobia are symbiotic soil bacteria able to intracellularly colonize legume nodule cells and form nitrogen-fixing symbiosomes therein. How the plant cell cytoskeleton reorganizes in response to rhizobium colonization has remained poorly understood especially because of the lack of an in vitro infection assay. Here, we report on the use of the heterologous HeLa cell model to experimentally tackle this question. We observed that the model rhizobium Sinorhizobium meliloti, and other rhizobia as...

Marchetti, Marta; Capela, Delphine; Poincloux, Renaud; Benmeradi, Nacer; Auriac, Marie-christine; Le Ru, Aure?lie; Maridonneau-parini, Isabelle; Batut, Jacques; Masson-boivin, Catherine

2013-01-01

320

Kurarinone promotes TRAIL-induced apoptosis by inhibiting NF-?B-dependent cFLIP expression in HeLa cells  

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This study was designed to investigate the effects of the prenylated flavonoid kurarinone on TNF-related apoptosis inducing ligand (TRAIL)-induced apoptosis and its underlying mechanism. A low dose of kurarinone had no significant effect on apoptosis, but this compound markedly promoted tumor cell death through elevation of Bid cleavage, cytochrome c release and caspase activation in HeLa cells treated with TRAIL. Caspase inhibitors inhibited kurarinone-mediated cell death, which indicates th...

Seo, Ok-won; Kim, Jung Hwan; Lee, Kwang-soon; Lee, Kyu-sun; Kim, Ji-hee; Won, Moo-ho; Ha, Kwon-soo; Kwon, Young-guen; Kim, Young-myeong

2012-01-01

 
 
 
 
321

Cannabidiol Inhibits Cancer Cell Invasion Via Upregulation Of Tissue Inhibitor Of Matrix Metalloproteinases-1  

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Abstract Although cannabinoids exhibit a broad variety of anticarcinogenic effects, their potential use in cancer therapy is limited by their psychoactive effects. Here we evaluated the impact of cannabidiol, a plant-derived non-psychoactive cannabinoid, on cancer cell invasion. Using Matrigel invasion assays we found a cannabidiol-driven impaired invasion of human cervical cancer (HeLa, C33A) and human lung cancer cells (A549) that was reversed by antagonists to both CB1 and CB2 r...

2010-01-01

322

Binding of 4'-aminomethyl 4,5',8-trimethyl psoralen to DNA, RNA and protein in HeLa cells and Drosophila cells  

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In Drosophila cells and HeLa cells treated with 4'-aminomethyl trioxsalen and ultraviolet light, this compound binds covalently to DNA and RNA. The maximum number of molecules bound to 103 base pairs in DNA is 60 and in RNA it is 20. In nuclei treated likewise the number of molecules bound to 103 base pairs in DNA can be as high as 376. When cells are irradiated in the frozen state the number of 4'-aminomethyl trioxsalen molecules bound per 103 base pairs in DNA is about 40 and in RNA about 20. DNA molecules from cells or nuclei treated with 4'-aminomethyl trioxsalen and ultraviolet light are highly crosslinked and appear as loops interspersed by double stranded regions when analyzed in the electron microscope under denaturing conditions. The loop sizes are heterogenous and the fraction of double stranded regions increases to almost complete double-strandedness at high degrees of reaction. No secondary structures could be found in ribosomal RNA from Drosophila cells or HeLa cells after treatment with 4'-aminomethyl trioxsalen and ultraviolet light. In cells treated with 4'-aminomethyl trioxsalen and ultraviolet light the RNAase activity is increased considerably suggesting a release of lysosomal enzymes. 4'-aminomethyl trioxsalen and its photodecomposition products bind strongly to cellular proteins. (Auth.)

1979-07-26

323

Silencing cytokeratin 18 gene inhibits intracellular replication of Trypanosoma cruzi in HeLa cells but not binding and invasion of trypanosomes  

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Full Text Available Abstract Background As an obligatory intracellular parasite, Trypanosoma cruzi, the etiological agent of Chagas' disease, must invade and multiply within mammalian cells. Cytokeratin 18 (CK18 is among the host molecules that have been suggested as a mediator of important events during T. cruzi-host cell interaction. Based on that possibility, we addressed whether RNA interference (RNAi-mediated down regulation of the CK18 gene could interfere with the parasite life cycle in vitro. HeLa cells transiently transfected with CK18-RNAi had negligible levels of CK18 transcripts, and significantly reduced levels of CK18 protein expression as determined by immunoblotting or immunofluorescence. Results CK18 negative or positive HeLa cells were invaded equally as well by trypomastigotes of different T. cruzi strains. Also, in CK18 negative or positive cells, parasites recruited host cells lysosomes and escaped from the parasitophorous vacuole equally as well. After that, the growth of amastigotes of the Y or CL-Brener strains, was drastically arrested in CK18 RNAi-treated cells. After 48 hours, the number of amastigotes was several times lower in CK18 RNAi-treated cells when compared to control cells. Simultaneous staining of parasites and CK18 showed that in HeLa cells infected with the Y strain both co-localize. Although the amastigote surface protein-2 contains the domain VTVXNVFLYNR previously described to bind to CK18, in several attempts, we failed to detect binding of a recombinant protein to CK-18. Conclusion The study demonstrates that silencing CK18 by transient RNAi, inhibits intracellular multiplication of the Y and CL strain of T. cruzi in HeLa cells, but not trypanosome binding and invasion.

de Mello Samanta M

2008-12-01

324

FAM129B/MINERVA, a Novel Adherens Junction-associated Protein, Suppresses Apoptosis in HeLa Cells*  

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A recent proteomics study identified FAM129B or MINERVA as a target of the MAP kinase (Erk1/2) signaling cascade in human melanoma cells. Phosphorylation of the protein was found to promote cell invasion and the dissociation of the protein from the cell-cell junctions. Suppression of apoptosis during metastasis is a prerequisite for the survival and spread of cancer cells. During apoptosis, the adherens junctions are disassembled as the dying cell retracts, and new contacts are formed between...

Chen, Song; Evans, Hedeel Guy; Evans, David R.

2011-01-01

325

Heat-shock pretreatment inhibits sorbitol-induced apoptosis in K562, U937 and HeLa cells.  

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The aim of this study was to determine whether heat-shock pretreatment exerted a protective effect against sorbitol-induced apoptotic cell death in K562, U937 and HeLa cell lines and whether such protection was associated with a decreased cytochrome c release from mithocondria and a decreased activation of caspase-9 and -3. Following heat-shock pretreatment (42 +/- 0.3 degrees C for 1 hr), these cell lines were exposed to sorbitol for 1 hr. Apoptosis was evaluated by DNA fragmentation, whereas caspase-9,-3 activation, cytochrome c release and heat-shock protein70 (HSP70) were assayed by Western Blot. Sorbitol exposure-induced apoptosis in these different cell lines with a marked activation of caspase-9 and caspase-3, whereas heat-shock pretreatment before sorbitol exposure, induced expression of HSP70 and inhibited sorbitol-mediated cytochrome c release and subsequent activation of caspase-9 and caspase-3. Similarly, overexpression of HSP70 in the three cell lines studied prevented caspase-9 cleavage and activation as well as cell death. Furthermore, we showed that the mRNA expression of iNOS decreased during both the heat-shock treatment and heat-shock pretreatment before sorbitol exposure. By contrast, the expression of Cu-Zn superoxide dismutase (SOD) and Mn-SOD proteins increased during heat-shock pretreatment before sorbitol exposure. We conclude that, heat-shock pretreatment protects different cell lines against sorbitol-induced apoptosis through a mechanism that is likely to involve SOD family members. PMID:19598258

Marfe, Gabriella; Pucci, Bruna; De Martino, Luisa; Fiorito, Filomena; Di Stefano, Carla; Indelicato, Manuela; Aventaggiato, Michele; Russo, Matteo A; Tafani, Marco

2009-11-01

326

Phytate decreases oxidative damage caused by labile forms of iron in solution, blood plasma and in HeLa cells  

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Full Text Available SciELO Brazil | Language: English Abstract in portuguese Fitato (PHYT, mio-inositol 1,2,3,4,5,6-hexakisfosfato) é um produto natural com forte efeito sobre a biodisponibilidade de minerais, especialmente o ferro. Neste trabalho, investigamos os efeitos antioxidantes do PHYT em modelos de transtornos de sobrecarga de ferro (ferro lábil plasmático e reserva [...] tório de ferro lábil). PHYT apresentou um efeito antioxidante considerável, com a vantagem de ser permeável às células e ser um constituinte normal da dieta humana. Nossos resultados sugerem que o PHYT pode auxiliar as defesas do organismo contra estresse induzido por sobrecarga de ferro. Abstract in english Phytate (PHYT, myo-inositol 1,2,3,4,5,6-hexakisphosphate) is a natural product with strong effect on the bioavailability of minerals, especially iron. In this work, we investigated the antioxidant effects of PHYT on models of iron overload disorders (labile plasma iron and labile iron pool) both in [...] solution and in HeLa cells. PHYT has a considerable antioxidant effect, with the benefit of being cell permeant and a normal constituent of human diet. Our results suggest that PHYT may assist organism defenses against iron-overload stress.

Frederico A., Schleh; Orlando, Chiarelli-Neto; Mayara N., Fontes; Renato, Najjar; Breno P., Espósito.

327

Conformational change in the floor of the human rhinovirus canyon blocks adsorption to HeLa cell receptors.  

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A series of eight antiviral compounds complexed with human rhinovirus 14 (HRV-14) were previously shown to displace segments of polypeptide chains in the floor of the "canyon" by as much as 0.45 nm in C-alpha positions from the native conformation (J. Badger, I. Minor, M. J. Kremer, M. A. Oliveira, T. J. Smith, J. P. Griffith, D. M. A. Guerin, S. Krishnaswamy, M. Luo, M. G. Rossman, M. A. McKinlay, G. D. Diana, F. J. Dutko, M. Fancher, R. R. Rueckert, and B. A. Heinz, Proc. Natl. Acad. Sci. USA 85:3304-3308, 1988). Because the canyon is thought to serve as the viral receptor-binding site (M. G. Rossmann, E. Arnold, J. W. Erickson, E. A. Frankenberger, J. P. Griffith, H. J. Hecht, J. E. Johnson, G. Kamer, M. Luo, A. G. Mosser, R. R. Rueckert, B. Sherry, and G. Vriend, Nature [London] 317:145-153, 1985; M. G. Rossmann and R. R. Rueckert, Microbiol. Sci. 4:206-214, 1987), these compounds were assessed for their ability to block adsorption of HRV-14 to HeLa cell membrane receptors. In parallel experiments, the compounds were assessed directly for antiviral activity in an in vitro plaque reduction assay in intact HeLa cells. All eight compounds blocked the adsorption of 50% of HRV-14 at approximately the same concentration required to reduce the number of visible plaques by 50% (MIC). A structurally related compound which was inactive in the plaque reduction assay had no effect on HRV-14 binding. A drug-resistant mutant of HRV-14 (Leu-1188), which was less sensitive to the eight compounds in plaque reduction assays was similarly less sensitive in the adsorption assay. We propose that the conformational changes in the floor of the HRV-14 canyon induced by these compounds substantially decrease adsorption of the virion to its receptor. These results provide further evidence for the role of the HRV canyon in receptor binding. PMID:2539499

Pevear, D C; Fancher, M J; Felock, P J; Rossmann, M G; Miller, M S; Diana, G; Treasurywala, A M; McKinlay, M A; Dutko, F J

1989-05-01

328

Measurement of the lateral diffusion of human MHC class I molecules on HeLa cells by fluorescence recovery after photobleaching using a phycoerythrin probe.  

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The mobility of cell surface MHC class I molecules on HeLa cells was measured by fluorescence recovery after photobleaching (FRAP). The probe used for these studies was the phycobiliprotein R-phycoerythrin coupled to Fab fragments of a monoclonal antibody specific for human monomorphic MHC class I molecules. It was found that the recovery curves could be equally well fitted by either a random diffusion model with an immobile component or by an anomalous diffusion model. In the latter case, th...

Georgiou, George; Bahra, Sukhvinder S.; Mackie, Alan R.; Wolfe, Caroline A.; O Shea, Paul; Ladha, Shab; Fernandez, Nelson; Cherry, Richard J.

2002-01-01

329

Characterization of SAF-A, a novel nuclear DNA binding protein from HeLa cells with high affinity for nuclear matrix/scaffold attachment DNA elements.  

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We identified four proteins in nuclear extracts from HeLa cells which specifically bind to a scaffold attachment region (SAR) element from the human genome. Of these four proteins, SAF-A (scaffold attachment factor A), shows the highest affinity for several homologous and heterologous SAR elements from vertebrate cells. SAF-A is an abundant nuclear protein and a constituent of the nuclear matrix and scaffold. The homogeneously purified protein is a novel double stranded DNA binding protein wi...

Romig, H.; Fackelmayer, F. O.; Renz, A.; Ramsperger, U.; Richter, A.

1992-01-01

330

Dissecting the Nanoscale Distributions and Functions of Microtubule-End-Binding Proteins EB1 and ch-TOG in Interphase HeLa Cells  

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Recently, the EB1 and XMAP215/TOG families of microtubule binding proteins have been demonstrated to bind autonomously to the growing plus ends of microtubules and regulate their behaviour in in vitro systems. However, their functional redundancy or difference in cells remains obscure. Here, we compared the nanoscale distributions of EB1 and ch-TOG along microtubules using high-resolution microscopy techniques, and also their roles in microtubule organisation in interphase HeLa cells. The ch-...

Nakamura, Satoko; Grigoriev, Ilya; Nogi, Taisaku; Hamaji, Tomoko; Cassimeris, Lynne; Mimori-kiyosue, Yuko

2012-01-01

331

Down-regulation of the sodium pump following chronic exposure of HeLa cells and chick embryo heart cells to ouabain.  

Science.gov (United States)

1 HeLa cells and primary cultures of embryonic chick heart cells were grown in medium containing low concentrations of ouabain for 24 h. 2 Compared with normal cells, cells grown in ouabain have fewer free sodium pump sites, an increased intracellular sodium concentration and a decreased intracellular potassium concentration. The cells are able to maintain their intracellular ion contents because the remaining pump sites have an increased turnover rate. 3 When cells that have been chronically exposed to ouabain are returned to normal growth medium, the sodium pump site numbers increase; the recovery process begins within 6 to 8 h and is complete within 24 h. Recovery of pump site numbers is primarily dependent upon de novo protein synthesis since the protein synthesis inhibitor, cycloheximide, prevents recovery. PMID:7236988

Aiton, J F; Lamb, J F; Ogden, P

1981-06-01

332

Mechanism of derivation of radioresistance in HeLa cell population after repeated x-irradiation  

International Nuclear Information System (INIS)

The Radioresistant strain (X-8-5) was obtained from HeLa-SC population X-irradiated repeatedly for five times with 800 rad. The mean lethal dose (D0) was 196 rad for X-8-5 cells, while it was 166 rad for control HeLa-SC cells. The fraction of cells containing an unusually long acrocentric chromosome (LA 2) exclusively increased with increasing number of irradiation of HeLa-SC population. A clonal strain with LA 2 marker was isolated from X-8-5 population and named RC-355. Since the RC-355 cells were more resistant (D0 = 220 rad)than parental X-8-5 cells (D0 = 196 rad), it was suggested that the cells with LA 2 were responsible for the radioresistance of X-8-5 population. The RC-355 cells were further subjected to the analysis of Q-banded karyotypes and it was observed that 18 types of specific markers (rm 1-17 and LA 2) were included in RC-355 cells in addition to 12 types of markers observed in most of HeLa-SC cells. Since the analysis of Q-banded karyotypes of RC-355 cells showed that RC-355 specific markers were not produced by radiation-induced rearrangements of HeLa-SC chromosomes, because twelve kinds of HeLa-SC markers were presented in RC-355 cells without any change, it was concluded that a small number of cells with LA 2 marker were originally presented in the control population and the relative fraction of them occupied increased after irradiation. (author)

1982-01-01

333

Effect of doxorubicin on cell survival and micronuclei formation in HeLa cells exposed to different doses of gamma-radiation  

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Purpose: The present study was undertaken to obtain an insight into the combined effects of doxorubicin with radiation on the cell survival and micronuclei induction in HeLa cells. Material and Methods: HeLa S3 cells were allowed to grow till they reached plateau phase, inoculated with 10 {mu}g/ml doxorubicin hydrochloride and then exposed to 0, 0.5, 1, 2 and 3 Gy {gamma}-radiation. Clonogenicity of cell was measured using the colony forming assay, micronuclei formation using the micronucleus assay. Results: The treatment of HeLa cells with doxorubicin (adriamycin) for 2 hours before exposure to different doses of {gamma}-radiation resulted in a significant and dose-dependent decline in the cell survival and cell proliferation when compared to the PBS+irradiation group. Conversely, the frequency of micronuclei increased in a dose-related manner in both the PBS+irradiation and doxorubicin+irradiation groups. The pretreatment of HeLa cells with doxorubicin before irradiation to various doses of {gamma}-rays resulted in a significant elevation in the frequency of micronuclei when compared with the concurrent PBS+irradiation group. The dose-response relationship for both PBS+irradiation and doxorubicin+irradiation groups was linear. The correlation between cell survival and micronuclei induction was also determined for PBS or doxorubicin+irradiation group, where the clonogenicity of cells declined with the increase in micronuclei formation. The correlation between cell survical and micronuclei induction was linear quadratic for both PBS+irradiation and doxorubicin+irradiation groups. Conclusion: From our study it can be concluded that combination treatment with doxorubicin and radiation increased the genotoxic effect of the either treatment given alone. (orig.) [German] Hintergrund: Der kombinierte Effekt einer Doxorubicingabe mit Bestrahlung auf das Zellueberleben und die Mikronukleiinduktion wurde an HeLa-Zellen untersucht. Material und Methoden: HeLa-S3-Zellen in der Plateauphase wurden mit 10 {mu}g/ml Doxorubicinhydrochlorid inokuliert und dann einer {gamma}-Bestrahlung von 0, 0,5, 1,2 und 3 Gy ausgesetzt. Die Klonogenitaet der Zellen wurde mit dem Koloniebildungstest, die Bildung von Mikronuklei mit Hilfe des Mikronukleusassays untersucht. Ergebnisse: Die Behandlung von HeLa-Zellen mit Doxorubicin fuer zwei Stunden vor einer {gamma}-Bestrahlung mit verschiedenen Dosen resultierte in einer signifikanten und dosisabhaengigen Abnahme des Zellueberlebens und der Zellproliferation im Vergleich zu einer nur bestrahlten Kontrollgruppe. Die Haeufigkeit der Mikronukleusbildung hingegen nahm in einer dosisabhaengigen Weise sowohl fuer allein bestrahlte wie mit Doxorubicin und Bestrahlung behandelte Gruppen zu. Die Vorbehandlung von HeLa-Zellen mit Doxorubicin vor Bestrahlung mit verschiedenen Dosen ergab einen signifikanten Anstieg der Haeufigkeit der Mikronuklei im Vergleich zu der nur bestrahlten Gruppe. Die Dosis-Wirkungs-Beziehung sowohl fuer die nur bestrahlte als auch fuer die mit Doxorubicin und Bestrahlung behandelte Gruppe war linear. Die Korrelation zwischen Zellueberleben und Mikronukleusinduktion wurde ebenso fuer die allein bestrahlte wie fuer die mit Doxorubicin und Bestrahlung behandelte Gruppe bestimmt; die Klonogenitaet der Zellen nahm dabei mit ansteigender Mikronukleusformation ab. Die Korrelation zwischen Zellueberleben und Mikronukleusinduktion war linear quadratisch sowohl fuer die nur bestrahlte als auch fuer die mit Doxorubicin und Bestrahlung behandelte Gruppe. Schlussfolgerung: Aus unserer Studie kann geschlossen werden, dass die Kombination aus Doxorubicin und Bestrahlung die genotoxische Wirkung der Einzelmodalitaeten erhoeht. (orig.)

Jagetia, G.C.; Nayak, V. [Kasturba Medical College, Manipal (India). Dept. of Radiobiology

2000-09-01

334

Elasticity Mapping Analysis of Apical Cell Periphery Actin Structures of Normal Fibroblasts and Cervical Cancer Cells  

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Full Text Available The cell mechanical features are largely regulated by actin cytokeleton. By analyzing the mechanical features, it is possible to evaluate the characteristics of the complicated actin cytoskeleton in diverse cell types. In this study, we examined the sub-membrane mechanical structures of normal fibroblasts TIG-1 cells, and cervical cancer Hela cells using local elasticity mapping method of atomic force microscope. Especially we aimed at clarifying the regulatory mechanisms of sub-membrane actin structures in these cells by activation of actomyosin formation using calyculin A. This technique revealed that TIG-1 and Hela cells bore clearly different sub-membrane mechanical structures. TIG-1 cells had aligned stiff filamentous structures, whereas Hela cells had crooked and relatively soft filaments. The surface stiffness of TIG-1 cells increased slightly by actomyosin formation due to stiffness increase of the aligned filamentous structures. On the other hand, the surface stiffness of Hela cells increased by actomyosin formation due to upregulation of the apical actin filaments. Therefore, the structural and regulatory differences of the apical actin filaments could be demonstrated by atomic force microscopy elasticity mapping analysis.

Takanori Kihara

2013-05-01

335

A Nucleic-Acid Hydrolyzing Single Chain Antibody Confers Resistance to DNA Virus Infection in HeLa Cells and C57BL/6 Mice.  

Science.gov (United States)

Viral protein neutralizing antibodies have been developed but they are limited only to the targeted virus and are often susceptible to antigenic drift. Here, we present an alternative strategy for creating virus-resistant cells and animals by ectopic expression of a nucleic acid hydrolyzing catalytic 3D8 single chain variable fragment (scFv), which has both DNase and RNase activities. HeLa cells (SCH7072) expressing 3D8 scFv acquired significant resistance to DNA viruses. Virus challenging with Herpes simplex virus (HSV) in 3D8 scFv transgenic cells and fluorescence resonance energy transfer (FRET) assay based on direct DNA cleavage analysis revealed that the induced resistance in HeLa cells was acquired by the nucleic acid hydrolyzing catalytic activity of 3D8 scFv. In addition, pseudorabies virus (PRV) infection in WT C57BL/6 mice was lethal, whereas transgenic mice (STG90) that expressed high levels of 3D8 scFv mRNA in liver, muscle, and brain showed a 56% survival rate 5 days after PRV intramuscular infection. The antiviral effects against DNA viruses conferred by 3D8 scFv expression in HeLa cells as well as an in vivo mouse system can be attributed to the nuclease activity that inhibits viral genome DNA replication in the nucleus and/or viral mRNA translation in the cytoplasm. Our results demonstrate that the nucleic-acid hydrolyzing activity of 3D8 scFv confers viral resistance to DNA viruses in vitro in HeLa cells and in an in vivo mouse system. PMID:24968358

Lee, Gunsup; Yu, Jaelim; Cho, Seungchan; Byun, Sung-June; Kim, Dae Hyun; Lee, Taek-Kyun; Kwon, Myung-Hee; Lee, Sukchan

2014-06-01

336

Hsp105 family proteins suppress staurosporine-induced apoptosis by inhibiting the translocation of Bax to mitochondria in HeLa cells  

International Nuclear Information System (INIS)

Hsp105 (Hsp105? and Hsp105?), major heat shock proteins in mammalian cells, belong to a subgroup of the HSP70 family, HSP105/110. Previously, we have shown that Hsp105? has completely different effects on stress-induced apoptosis depending on cell type. However, the molecular mechanisms by which Hsp105? regulates stress-induced apoptosis are not fully understood. Here, we established HeLa cells that overexpress either Hsp105? or Hsp105? by removing doxycycline and examined how Hsp105 modifies staurosporine (STS)-induced apoptosis in HeLa cells. Apoptotic features such as the externalization of phosphatidylserine on the plasma membrane and nuclear morphological changes were induced by the treatment with STS, and the STS-induced apoptosis was suppressed by overexpression of Hsp105? or Hsp105?. In addition, we found that overexpression of Hsp105? or Hsp105? suppressed the activation of caspase-3 and caspase-9 by preventing the release of cytochrome c from mitochondria. Furthermore, the translocation of Bax to mitochondria, which results in the release of cytochrome c from the mitochondria, was also suppressed by the overexpression of Hsp105? or Hsp105?. Thus, it is suggested that Hsp105 suppresses the stress-induced apoptosis at its initial step, the translocation of Bax to mitochondria in HeLa cells

2006-10-15

337

Heat induced protein denaturation in the particulate fraction of HeLa S3 cells: effect of thermotolerance.  

Science.gov (United States)

In this study we investigated the effect of heat on the proteins of the particulate fraction (PF) of HeLa S3 cells using electron spin resonance (ESR) and thermal gel analysis (TGA). ESR detects overall conformational changes in proteins, while TGA detects denaturation (aggregation due to formation of disulfide bonds) in specific proteins. For ESR measurements the -SH groups of the proteins were labelled with a maleimido bound spin label (4-maleimido-tempo). The sample was heated inside the ESR spectrometer at a rate of 1 degree C/min. ESR spectra were made every 2-3 degrees C between 20 degrees C and 70 degrees C. In the PF of untreated cells conformational changes in proteins were observed in three temperature stretches: between 38 and 44 degrees C (transition A, TA); between 47 and 53 degrees C (transition B, TB); and above 58 degrees C (transition C, TC). With TGA, using the same heating rate, we identified three proteins (55, 70, and 90 kD) which denatured during TB. No protein denaturation was observed during TA, while during TC denaturation of all remaining proteins in the PF occurred. When the ESR and TGA measurements were done with the PF of (heat-induced) thermotolerant cells, TA was unchanged while TB and TC started at higher temperatures. The temperature shift for the onset of these transitions correlated with the degree of thermotolerance that was induced in the cells. These results suggest that protection against heat-induced denaturation of proteins in the PF is involved in heat induced thermotolerance. PMID:1325981

Burgman, P W; Konings, A W

1992-10-01

338

Study on the mutation spectrum of hprt gene in HeLa MR cell induced by ?-ray  

International Nuclear Information System (INIS)

Normal HeLa MR cells were irradiated by 60Co ?-ray in the dose range of 0?8 Gy and the mutants of hprt gene were cloned in the 6-TG containing selective culture and assayed. Three natural spontaneous mutants and eighteen ?-ray induced hprt mutants were recovered. The mutation spectra of hprt exons in all of them were analyzed by polymerase chain reaction (PCR) using 8 pairs of hprt oligonucleotide primers to amplify hprt exons from cellular DNA extracts. The result showed that in the mutants induced by ?-ray, about 77.8% (14/18) of the hprt mutations were deleted, among which 2 were total deletion, the other 12 were partial deletion, and 22.2% (4/18), which did not show noticeable changes in the PCR pattern, were point mutation. The mutation degree of hprt exon deletion was found to be dependent on the irradiation dose. However, about 67%(2/3) point mutants and 33%(1/3) deleted mutants were showed in the natural mutants. Their mutation degree was far lower than that of the ?-ray induced mutants. These results might be beneficial for further study of the molecular mechanism of mutation induced by ?-ray irradiation as well as for the estimation of irradiation dose

1998-02-01

339

Cytotoxic and Apoptotic Potentials of Ganoderma lucidum and Curculigo pilosa on Human Cervical Adenocarcinoma Cell Line, HeLa  

Directory of Open Access Journals (Sweden)

Full Text Available Many African natural products have been hypothesized to have phytochemicals that makes them effective anti-tumour agents. This research study looks at two out of the numerous hypothesized medicinal plants-Curculigo pilosa and Ganoderma lucidum. Caspase-3, Neutral red and DNA fragmentation assays were carried out on HeLa cell lines cultured in Dulbecco’s Modified Eagles Medium (DMEM in (95% O2 + 5% CO2 at 35°C. The apoptotic, cytotoxic capacities and DNA fragmentation assays were carried out on the medicinal plants. Both plant samples were extracted in both organic (mixture of ethanol and ethylacetate in the ratio 50:50 and aqueous solution (mixture of methanol and distilled water in the ratio 70:30. It was observed that both plant samples had apoptotic effects but below 50% of comparative levels with the exception of the aqueous extract of Ganoderma lucidum which could pass as an antitumour agent (showing apoptotic effect above 50%. Conclusively, the aqueous extract of Ganoderma lucidum proves to be suitable for the development of an antitumour agent as shown by its apoptotic effect reported in this study.

James Ayorinde Babatunde

2013-01-01

340

RNA splicing products formed with isolated fractions from HeLa cells are associated with fast-sedimenting complexes  

International Nuclear Information System (INIS)

Three fractions (designated Ia, Ib, and II) have been isolated from HeLa cell nuclear extracts that are required for splicing of adenovirus and human ?-globin RNA transcripts in vitro. The incubation of two of the fractions (Ib and II) in the presence of ATP resulted in cleavage of precursor mRNA at the 5' splice site and formation of the intron-exon lariat. Addition of fraction Ia to the combination of Ib and II resulted in the formation of spliced RNA and the intron lariat 32P-labelled 40s ribosomes were utilized. When fraction II was incubated with precursor RNA in the presence of ATP and the resulting products were sedimented through sucrose gradients, a 30S complex was detected that contained precursor RNA. The combination of fractions Ib and II resulted in the production of a 55S complex that contained the 5' exon as a prominent RNA species. The combination of fractions I (containing Ia and Ib) and II resulted in the formation of the 55S complex and material sedimenting between 40 S and 20 S, in which the predominant RNA species was spliced RNA

1986-01-01

 
 
 
 
341

Basal PIR expression in HeLa cells is driven by NRF2 via evolutionary conserved antioxidant response element.  

Science.gov (United States)

Pirin, a product of the PIR gene, is an iron-binding protein acting as a transcriptional coregulator implicated in the regulation of the NF-?B-related transcription via interaction with RelA (p65), as well as BCL3 and NF-?B1 (p50) proteins. Alterations in pirin expression were observed in various tumors and under oxidative stress conditions. The aim of the present work was to analyze the regulation of the transcription of the human PIR gene. Using constructs containing a different sized PIR promoter and the luciferase reporter genes we found that in HeLa cells PIR transcription is mostly dependent on a highly conserved antioxidant response element located 281 bp downstream of the transcription start site. We have proved that the NRF2 transcription factor binds to this element in vivo and drives the basal PIR expression. We hypothesize that regulation of the PIR expression may constitute a mechanism by which NRF2 is able to modulate the activity of NF-?B and possibly other signaling pathways. PMID:24390086

Brzóska, Kamil; St?pkowski, Tomasz M; Kruszewski, Marcin

2014-04-01

342

Enhancement of Chlamydia trachomatis infectious progeny by cultivation of HeLa 229 cells treated with DEAE-dextran and cycloheximide.  

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The effects of DEAE-dextran and cycloheximide on the infection of HeLa 229 cells with Chlamydia trachomatis serotype G were studied in terms of the number of cells infected and the yield of infectious progeny per infected cell. Pretreatment of the host cells with DEAE-dextran resulted in an increase in the number of infected cels but had no significant effect on the yield of infectious progeny per infected cell (burst size). In contrast, the addition of cycloheximide to the medium of infected...

1984-01-01

343

Effects of activated aflatoxin B"1 and caffeine on DNA replicon initiation in HeLa cells  

International Nuclear Information System (INIS)

Afatoxin B"1 (AFB"1) is activated by a rat microsomal extract (S-9) to form a product that inhibits DNA synthesis in HeLa cells. At 10"-_7 M, AFB"1 inhibited initiation of replicons, as shown in alkaline sucrose gradient profiles 30 min after incubation with the drug. Ninety minutes later, the profile of treated cells was similar to that of control, but 4 h later there was another effect on replicon initiation. At 10"-_6 M, the inhibition of initiation was greater than at 10"-_7 M and increased progressively. Four hours after removal of the drug, the gradient profile showed low amounts of radioactivity in all size classes of DNA. When cells were incubated in medium containing caffeine (2 mM) even as late as 60 min after incubation with AFB"1, the inhibition of replicon initiation was prevented. If caffeine was later removed from the medium, replicon initiation was then inhibited. At 10"-_7 M or 10"-_6 M, AFB"1 had little immediate effect on chain elongation, but at 10"-_5 M, the gradient profiles showed an accumulation of low molecular weight DNA molecules, with no radioactivity in the region of high molecular weight DNA, owing to a block to chain elongation; this was not affected by caffeine. These results suggest that AFB"1 induces damage that changes the fonformation of chromatin so that initiation of new replicons cannot occur; in the presence of caffeine this change does not occur and DNA replication is not inhibited

1981-01-01

344

Adherence to HeLa cells, typing by killer toxins and susceptibility to antifungal agents of Candida dubliniensis strains / Adesão a células HeLa, tipagem pelas toxinas "killer" e sensibilidade a antifúngicos de cepas de Candida dubliniensis  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese O objetivo do presente trabalho foi avaliar o comportamento de cepas de Candida dubliniensis recuperadas de pacientes HIV+ e com AIDS por meio da pesquisa de capacidade de adesão a células HeLa, susceptibilidade a toxinas "Killer" e resistência in vitro a antifúngicos (eTest® AB Biodisk, Solna, Suéc [...] ia). O ensaio de adesão foi fortemente aderente para a amostra padrão ATCC 777, e aderente para os demais isolados. Os testes de tipagem das amostras frente às cepas-padrão produtoras de toxinas "Killer" mostraram dois biótipos diferentes dos 9 isolados estudados: 888 e 688. Somente o biótipo 688 (ATCC 777) de C. dubliniensis foi sensível à toxina K2. Houve correlação inversa significativa entre adesão e sensibilidade a toxinas "killer" (r = -0,8525 - p = 0,0035). Em relação à pesquisa de resistência a antifúngicos, as amostras de C. dubliniensis foram sensíveis ao fluconazol, itraconazol, cetoconazol, voriconazol, à flucitosina e anfotericina B. Com exceção da amostra ATCC 777, todas as demais mostraram comportamento similar. Abstract in english The aim of this study was to evaluate the adherence capability to HeLa cells, the susceptibility to killer toxins and the in vitro susceptibility to antifungal agents (eTest? method - AB Biodisk, Solna, Sweden) of 9 Candida dubliniensis isolates recovered from HIV+ and AIDS patients. The adherence t [...] est was strongly positive for strain ATCC 777 and positive for all other strains. Typing by killer toxins revealed two different biotypes among the 9 isolates studied: 888 and 688. Only biotype 688 (ATCC 777) was susceptible to the K2 toxin. There was a significant inverse correlation between adherence and killer toxin susceptibility (r = -0.8525 - p = 0.0035). No strains presented resistance to fluconazole, itraconazole, ketoconazole, voriconazole, flucytosine or amphotericin-B. With the exception of ATCC 777, all the other isolates presented similar behavior.

Silva, Gismari Miranda da; Silveira, Fernando Ricardo Xavier da; Pires, Maria de Fátima Costa.

345

MODULATION OF SOME MEMBRANARY AND METABOLIC PROCESSES OF HeLa TUMORAL CELLS BY ELECTROMAGNETIC FIELDS OF LOW FREQUENCY AND INTENSITY  

Directory of Open Access Journals (Sweden)

Full Text Available The present paper represents the result of a study on the HeLa neoplastic cells’ membranary and metabolic reactivity to the action of non-ionizing electromagnetic field of continuous or discontinuous type. The in vitro shortlasting electromagnetic field exposure of the human tumoral cells has induced a significant modulation of the membrane Na + -K + -ATP-ase activity, comparatively with the control level. The same electromagnetic treatment has also conditioned an alteration of the control metabolic profile. Thus, we have appreciated a stimulation of the glycogenesis, lipogenesis, proteinsynthesis and nucleic acids biosynthesis, correlated with an intensification of the intracellular consumption – on catabolic and/or anabolic pathways – of the glucose, lactic acid, free fatty acids and aminoacids. The direction and amplitude of the modifications of the HeLa cellular processes are dependent to the type of electromagnetic field application.

Hellen Rotinberg

2007-12-01

346

Instant Response of Live HeLa Cells to Static Magnetic Field and Its Magnetic Adaptation  

CERN Document Server

We report Static Magnetic Field (SMF) induced altered sub-cellular streaming, which retains even after withdrawal of the field. The observation is statistically validated by differential fluorescence recovery after photo-bleaching (FRAP) studies in presence and absence of SMF, recovery rate being higher in presence of SMF. This instant magneto-sensing by live cells can be explained by inherent diamagnetic susceptibility of cells and alternatively by spin recombination, e.g., by the radical pair mechanism. These arguments are however insufficient to explain the retention of the SMF effect even after field withdrawal. Typically, a relaxation time scale at least of the order of minutes is observed. This long duration of the SMF effect can be explained postulating a field induced coherence that is followed by decoherence after the field withdrawal. A related observation is the emergence of enhanced magnetic susceptibility of cells after magnetic pre-incubation. This implies onset of a new spin equilibrium state a...

Raja, Sufi O

2014-01-01

347

Syzygium cumini inhibits growth and induces apoptosis in cervical cancer cell lines: a primary study  

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Cervical cancer is common among women in the Indian subcontinent and the incidences and death rates are gradually increasing over the years. Several dietary phytochemicals have been reported to have growth inhibitory and apoptotic effect on HeLa and other cervical cell lines. In this study, using Hoechst 33342 staining, MTT, Annexin V-FLUOS/PI and TUNEL assays we demonstrated that Syzygium cumini extract inhibits the growth and induces apoptosis in HeLa and SiHa cervical cancer cell lines in ...

2008-01-01

348

Syzygium cumini inhibits growth and induces apoptosis in cervical cancer cell lines: a primary study.  

Science.gov (United States)

Cervical cancer is common among women in the Indian subcontinent and the incidences and death rates are gradually increasing over the years. Several dietary phytochemicals have been reported to have growth inhibitory and apoptotic effect on HeLa and other cervical cell lines. In this study, using Hoechst 33342 staining, MTT, Annexin V-FLUOS/PI and TUNEL assays we demonstrated that Syzygium cumini extract inhibits the growth and induces apoptosis in HeLa and SiHa cervical cancer cell lines in a dose- and time-dependent manner. The phytochemical, its mode of action and safety issues are yet to be determined. PMID:22275971

Barh, D; Viswanathan, G

2008-01-01

349

Evaluation of Cytotoxic Effects of Dichloromethane Extract of Guduchi (Tinospora cordifolia Miers ex Hook F & THOMS) on Cultured HeLa Cells  

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Extracts of Tinospora cordifolia (TCE) have been shown to possess anti-tumor properties, but the mechanism of the anti-tumor function of TCE is poorly understood. This investigation elucidates the possible mechanism underlying the cytotoxic effects of dichlormethane extracts of TCE, after selecting optimal duration and concentration for treatment. HeLa cells were exposed to various concentrations of TCE, which has resulted in a concentration-dependent decline in the clonogenicity, glutathione...

2006-01-01

350

Transcription of adenovirus and HeLa cell genes in the presence of drugs that inhibit topoisomerase I and II function.  

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The requirements for topoisomerases in transcription of adenovirus and HeLa cell genes were analyzed using drugs that specifically inhibit either topoisomerases I or II. Cleavage of viral DNA by topoisomerases in the presence of either camptothecin or VM26 was used to determine drug concentrations that led to maximal inhibition of ligation in the cleavage and ligation step of topoisomerase I or II respectively. Inhibition of topoisomerase II with VM26 did not cause a direct reduction in trans...

1990-01-01

351

Acylated simian virus 40-specific proteins in the plasma membrane of HeLa cells infected with adenovirus 2-simian virus 40 hybrid virus Ad2+ND2  

Energy Technology Data Exchange (ETDEWEB)

HeLa cells infected with the adenovirus 2-simian virus 40 (Ad2+SV40) hybrid virus Ad2+ND2 were labeled with either (/sup 35/S)methionine or (/sup 3/H)palmitate and fractionated into cytoplasmic, nuclear, and plasma membrane fractions. Analysis of these fractions by sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that the SV40-specific proteins in the plasma membrane fraction were specificially acylated.

Klockmann, U.; Deppert, W.

1983-04-30

352

Cytotoxic and Apoptotic Potentials of Ganoderma lucidum and Curculigo pilosa on Human Cervical Adenocarcinoma Cell Line, HeLa  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Many African natural products have been hypothesized to have phytochemicals that makes them effective anti-tumour agents. This research study looks at two out of the numerous hypothesized medicinal plants-Curculigo pilosa and Ganoderma lucidum. Caspase-3, Neutral red and DNA fragmentation assays were carried out on HeLa cell lines cultured in Dulbecco’s Modified Eagles Medium (DMEM) in (95% O2 + 5% CO2) at 35°C. The apoptotic, cytotoxic capacities and DN...

Samuel Titilola Aderonke; Selina, Odesanmi O.; James Ayorinde Babatunde; Tafida Mundi; Magbagbeola, Olubunmi A.

2013-01-01

353

Testicular Germ Cell Cancer  

Science.gov (United States)

More than 90 percent of testicular cancer start in the germ cells, which are cells in the testicles and develop into sperm. This type of cancer is known as testicular germ cell cancer. Testicular germ cell cancer can be classified as either seminomas or nonseminomas, whose cells have different appearances under a microscope.1 Another difference is that nonseminomas typically grow and spread more quickly than seminomas.

354

Targeting SPARC by lentivirus-mediated RNA interference inhibits cervical cancer cell growth and metastasis  

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Abstract Background Secreted protein acidic and rich in cysteine (SPARC), a calcium-binding matricellular glycoprotein, is implicated in the progressions of some cancers. However, no information has been available to date regarding the function of SPARC in cervical cancer cell growth and metastasis. Methods In this study, we isolated and established high invasive subclones and low invasive subclones from human cervical cancer cell lines HeLa and SiHa by the limi...

Chen Jie; Shi Dehuan; Liu Xiaoyan; Fang Shuang; Zhang Jie; Zhao Yueran

2012-01-01

355

Ursolic Acid Induces Apoptosis Through Mitochondrial Intrinsic Pathway and Suppression of ERK1/2 MAPK in HeLa Cells.  

Science.gov (United States)

Ursolic acid (UA), a natural pentacyclic triterpenoid compound, has been demonstrated to induce apoptosis in various tumors. The aim of the present study was to elucidate the molecular mechanisms of UA-induced apoptosis in HeLa cells. Here, we reported that UA induced apoptosis through the mitochondrial intrinsic pathway in HeLa cells, as shown by release of cytosol cytochrome c, activation of caspase-9 and -3, reduction of Bcl-2 and Bcl-xL, and increase of Bax and Bak. UA down-regulated the phosphorylation of ERK1/2 and p38, whereas phosphorylation of JNK was unchanged. The roles of ERK1/2 and p38 were further confirmed using the ERK1/2 inhibitor (U0126) and p38 inhibitor (SB203580). U0126 markedly increased UA-induced the Bax/Bcl-2 ratio, the increase of cytosol cytochrome c, and the levels of cleaved caspase-3, but SB203580 had little effects on the above characters, suggesting the ERK1/2 signaling pathway is required for apoptosis. Furthermore, UA up-regulated DUSP 1, 2, 4, 5, 6, 7, 9, and 10 mRNA expressions, which may be a clue for the role of dephosphorylation of ERK1/2 and p38. These data suggested that the apoptotic mechanism of UA treatment in HeLa cells was through the mitochondrial intrinsic pathway and closely associated with the suppression of the ERK1/2 signaling pathway. PMID:24881958

Li, Yanhong; Lu, Xueying; Qi, Hongxue; Li, Xiaobo; Xiao, Xiangwen; Gao, Jianfeng

2014-06-19

356

Alterations of transcription and translation in HeLa cells exposed to amino acid analogs.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Amino acid analogs, like other effectors of the stress response, induce in mammalian cells the same gene products that are induced upon heat shock; incorporation of the analog into protein is required for induction. We show here that induction by analogs involves controls operating at the levels of both transcription and translation. The electrophoretic patterns of newly made mRNAs simplify with time such that the putative stress protein mRNAs are the only species transported from the nucleus...

Thomas, G. P.; Mathews, M. B.

1984-01-01

357

Reversibility of Membrane N-Glycome of HeLa Cells upon Treatment with Epigenetic Inhibitors  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Glycans are essential regulators of protein function and are now in the focus of research in many physiological and pathophysiological processes. There are numerous modes of regulating their biosynthesis, including epigenetic mechanisms implicated in the expression of glyco-genes. Since N-glycans located at the cell membrane define intercellular communication as well as a cellular response to a given environment, we developed a method to preferentially analyze this fraction of glycans. The me...

2013-01-01

358

Development of kinetochores during early mitosis in HeLa cells and the stability of a trilaminar kinetochore structure  

Energy Technology Data Exchange (ETDEWEB)

The ultrastructure of the kinetochore varies during the course of mitosis. This variation is considered to be a progressive maturation process that may correlate with kinetochore function. We examined the development of kinetochore structures on condensing prophase chromosomes in HeLa cells. Usually, one accepts the three laminar structure of the kinetochore of a condensed metaphase chromosome to reflect the general state and to be a prerequisite for microtubule attachment. The study of developing kinetochores in mitotic cells revealed unquestionable exceptions from this dogma. At the onset of chromatin condensation, indicating the beginning of prophase, the nuclear envelope and the nucleoli are still intact. First indications for kinetochore formation, documented as clouds of a fibrous material or as the {open_quotes}kinetochore ball structure{close_quotes} can be identified along or close to distinct heterochromatic, condensed chromatin elements. These fibrils extend from or interdigitate with the folding chromatin. Enhanced density of the ball structure represents a condensation process. Concomitantly a segregation of a plate structure is detected, occuring at a distal zone but obviously not at the end of the fibrils. This differentiation into kinetochore plate and corona more or less coincides with two other important prophase processes: the first tangential contacts to single microtubules and the mitotic breakdown of the nuclear envelope. But evidence will be discussed that these early prophase features not necessarily depend on each other and that the formation of a characteristic, trilaminar kinetochore structure does not guarantee its function. Conversely, structurally incomplete kinetochores can establish microtubular contacts for the attachment of chromosomes into the spindle.

Schroeter, D.; Paweletz, N.; Finze, E.M.; Kiesewetter, U.L. [German Cancer Research Center, Heidelberg (Germany)

1993-12-31

359

SMILE silencing and PMA activation gene networks in HeLa cells: comparison with kidney transplantation gene networks.  

Science.gov (United States)

Recent findings indicated that the SMILE gene may be involved in kidney graft operational tolerance in human. This gene was found to be up-regulated in blood from patients with a well functioning kidney transplant in the absence of immunosuppression compared to other transplanted recipients with clinically different status. A microarray study of SMILE knock-down and phorbol 12-myristate 13-acetate (PMA) activation in HeLa cells was herein compared to our earlier analysis based on microarray data of kidney allograft tolerance and rejection in humans and in a rat model of allograft transplantation to determine possible new genes and gene networks involved in kidney transplantation. The nearest neighbors at the intersection of the SMILE knock-down network with the human tolerance/rejection networks are shown to be NPHS1 and ARRB2, the former (Nephrin) being involved in kidney podocyte function, and the decrease of the latter (Arrestin ?2) being recently shown to be involved in monocyte activation during acute kidney allograft rejection in rat. Moreover, another one of the neighbors at the intersection of SMILE network and tolerance/rejection networks is XBP-1, that we report previously to be increased, at a transcript level, after ER stress in SMILE silenced cells. Finally, in this study, we also show that topological properties (both local and global) of joint SMILE knock-down network-tolerance/rejection networks and joint PMA activation network-tolerance/rejection networks in rat and human are essentially different, likely due to the inherent nature of the gene SMILE and the mitogen PMA, that do not act the same way on genes and do not interfere the same way on networks. We also show that interestingly SMILE networks contain more feed-forward loop (FFL) motifs and thus SMILE calls for a more fine-tuned genetic regulation. PMID:22134986

Racapé, M; Bragazzi, N; Sivozhelezov, V; Danger, R; Pechkova, E; Duong Van Huyen, J P; Soulillou, J P; Brouard, S; Nicolini, C

2012-06-01

360

Transporter Molecules influence the Gene Expression in HeLa Cells  

Science.gov (United States)

Progresses in biology and pharmacology led to highly specific bioactive substances, but their poor bioavailability at the site of action is a result of their physico-chemical properties. Various design approaches for transport carrier molecules facilitating the cellular entry of bioactive substances could help to reach their molecular target in cells and tissues. The transfer efficacy and the subsequent pharmacological effects of the cargo molecules are well investigated, but the investigations of effects of the carrier molecules themselves on the target cells or tissues remain necessary. A special attention should be paid to the differential gene expression, particularly in the interpretation of the data achieved by highly specific active pharmaceutical products. After application of transmembrane transport peptides, particularly the pAnt and also the HIV-1 Tat, cells respond with a conspicuous altered gene expression of at least three genes. The PKN1 gene was induced and two genes (ZCD1 and BSG) were slightly repressed. The genes and the chromosomes are described, the moderate differential gene expression graphed, and the ontology is listed.

Waldeck, Waldemar; Pipkorn, Ruediger; Korn, Bernhard; Mueller, Gabriele; Schick, Matthias; Toth, Katalin; Wiessler, Manfred; Didinger, Bernd; Braun, Klaus

2009-01-01

 
 
 
 
361

Transporter molecules influence the gene expression in HeLa cells.  

Science.gov (United States)

Progresses in biology and pharmacology led to highly specific bioactive substances, but their poor bioavailability at the site of action is a result of their physico-chemical properties. Various design approaches for transport carrier molecules facilitating the cellular entry of bioactive substances could help to reach their molecular target in cells and tissues. The transfer efficacy and the subsequent pharmacological effects of the cargo molecules are well investigated, but the investigations of effects of the carrier molecules themselves on the target cells or tissues remain necessary. A special attention should be paid to the differential gene expression, particularly in the interpretation of the data achieved by highly specific active pharmaceutical products. After application of transmembrane transport peptides, particularly the pAnt and also the HIV-1 Tat, cells respond with a conspicuous altered gene expression of at least three genes. The PKN1 gene was induced and two genes (ZCD1 and BSG) were slightly repressed. The genes and the chromosomes are described, the moderate differential gene expression graphed, and the ontology is listed. PMID:19214198

Waldeck, Waldemar; Pipkorn, Ruediger; Korn, Bernhard; Mueller, Gabriele; Schick, Matthias; Tóth, Katalin; Wiessler, Manfred; Didinger, Bernd; Braun, Klaus

2009-01-01

362

Mitotic block and delayed lethality in HeLa epithelial cells exposed to Escherichia coli BM2-1 producing cytotoxic necrotizing factor type 1.  

Science.gov (United States)

The cytopathic effect (CPE) of Escherichia coli producing cytotoxic necrotizing factor type 1 (CNF1) was investigated by using a human epithelial cell (HeLa) model of infection with CNF1-producing E. c