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1

Radiation sensitization by dihydroartemisinin on human HeLa cells of cervical cancer  

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Objective: To investigate the radiosensitizing effects of dihydroartemisinin (DHA) on human HeLa cells of cervical cancer irradiated by X rays. Methods: Cell growth kinetics was determined using MTF assay. Cell survival was analyzed by elonogenic assay. The change of cell cycle and apeptosis was measured by flow cytometry. Results: Dihydroartemisinin inhibited the growth of HeLa cells of human cervical cancer and showed a dose-dependent and time-dependent manner. Dihydroartemisinin (20 ?mol/L) showed the radiosensitizing effects on HeLa cells, and the sensitizing enhancement ratio (SER) was 1.47. Dihydroartemisinin abrogated radiation-induced G2 arrest of the tested HeLa cells, the G2 ratio of medicine + radiation group dechned from 73.58% to 48.31%. Dihydroartemisinin enhanced the apoptosis of HeLa cells by X-irradiation, the apoptosis rates of medicine + radiation group significantly increased from 29.46%, 48.04%, 70.21% to 45.79%, 66.36% and 79.58%, respectively for 2, 4 and 6 Gy. Conclusions: Dihydroartemisinin could increase the radiosensitivity of HeLa cells of human cervical cancer. Abrogation of radiation-induced C2 arrest could be part of the mechanism. (authors)

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

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

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

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

Liu Xianting

2010-01-01

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Radiation sensitization by CAPE on human HeLa cells of cervical cancer  

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Objective: To study the radiosensitizing effect of caffic acid phenethyl ester (CAPE) on human cervical cancer HeLa cells. Methods: MTT assay was used to measure the relation between the inhibition effect and CAPE concentrations by CAPE with different concentrations on HeLa cells for 24 hours. HeLa cells were divided into the control and experimental groups, both of which were given 0, 2, 4, 6 and 8 Gy of 60Co ?-irradiation, respectively. The cell clones were counted. Meanwhile HeLa cells were divided into the control, CAPE, irradiation and combination groups. Flow cytometric analysis was adopted to detect the changes of cell cycle distribution induced by CAPE. Results: The inhibition rate of CAPE acting on Hela cells increased with concentrations (F=126. 49 ? 3654.88, P0) (1.45 and 1.82 Gy) and the quasi-threshold dose (Dq) (1.89 and 3.21 Gy) of HeLa cells in experimental group decreased comparing with control group, SER was 1.26. Compared with the sole irradiation group, cells in G2/M phase of the CAPE group and the sole irradiation group increased (P2/M arrest and may be related to the inhibition of the sub-lethal damage repair. (authors)

5

Study of radiation sensitization of artesunate on human HeLa cells of cervical cancer  

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Objective: To investigate the radiosensitizing effects of artesunate on human HeLa cells of cervical cancer in vitro. Methods: Hela cells irradiated with 60Co ?-rays. The dose rate was 0.635 Gy/min and the radiation dose was 0, 1, 2, 4, 6 Gy, respectively. The anti-proliferation activities of artesunate on HeLa cells were evaluated with MTT assay, to determine the most appropriate drug concentration. The effect of radiosensitivity was observed by using clonogenic assay. The single-hit multi-target model was used to plot the HeLa cell's dose-survival curve, to calculate mean lethal dose, quasi-threshold dose and sensitization enhancement rate, and to evaluate its radiosensitization effect. The apoptosis was analyzed with flow cytometry (FCM) to further test the radiation sensitization of artesunate on HeLa cells. Results: The inhibition of artesunate on HeLa cells increased with concentration. In radiation group, the cell cloning efficiency were 91.67%, 82.02%, 58.06%, 25.01%, respectively, and in artesunate (2.0 ?mol/L) + radiation group, the cell cloning efficiency were 74.93%, 60.53%, 22.38%, 5.05%. In radiation group and artesunate (2.0 ?mol/L) + radiation group, the mean lethal dose (D0) was 2.95 and 2.07 Gy, respectively, while the qusai-threshold dose (Dq) were 2.01 and 1.24 Gy, respectively, and SER was 1.43. Compared with 2 and 6 Gy radiation group, the apoptosis rate of drug + radiation group increased from 12.26%, 40.08% to 22.71%, 59.92. Conclusions: The inhibiting effect of artesunate on HeLa cells is concentration-dependent. Artesunate has radiosensitizing effect on HeLa cells in vitro. (authors)

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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.

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Combined treatment of ionizing radiation with genistein on cervical cancer HeLa cells  

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The anticancer agent genistein inhibits cell growth of tumor cell lines from various malignancies. In our study, we investigated the effectiveness of combined treatment of ionizing radiation (IR) with genistein on cervical HeLa cells and its possible mechanism. It was found that the inhibitory rate in cells with combined treatment was significantly higher than that of the cells treated with IR or genistein alone. After treatments of IR (4 Gy) combined with genistein (40 ?mol/L), the apoptotic index of the cells was significantly increased and the cells were arrested in the G2/M phase. Survivin mRNA expression increased after IR (4 Gy), while it significantly decreased after combined treatment. These findings indicated that genistein enhanced the radiosensitivity of cervical cancer HeLa cells, and the mechanisms for this action might include increase of apoptosis, decrease of survivin expression, and prolongation of cell cycle arrest. (author)

8

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

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

Liu Junye

2012-08-01

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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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Effects of Smac gene over-expression on radiotherapeutic sensitivity of cervical cancer cell line HeLa  

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Objective: To study the effects of extrinsic Smac gene transfection and its over-expression on radiotherapeutic sensitivity of cervical cancer cells, in order to explore a novel strategy for ameliorating radiotherapy of cervical cancer. Methods: After Smac gene was transferred into cells of cervical cancer cell line HeLa, the subclone cells were obtained by persistent G418 selection. Cellular Smac gene expression was determined by RT-PCR and Western blot. After treatment with X-ray irradiation, cellular growth activity in vitro was investigated by MTT colorimetry. Cellular apoptosis and its rate were determined by electron microscopy, Annexin V-FITC and propidium iodide staining flow cytometry. Cellular Caspase-3 protein expression and its activity were assayed by Western blot and colorimetry. Results: RT-PCR and Western blot demonstrated that Smac mRNA and protein levels of HeLa/Smac cells, the selected subclone cells of cervical cancer cell line, were significantly higher than those of HeLa cells (P<0.01). After treated with 8 Gy X-ray irradiation, growth activity of HeLa/Smac cells reduced by 10.19%(P<0.01), as compared with that of HeLa cells. Partial HeLa/Smac cancer cells presented characteristic morphological changes of apoptosis under electron microscope, with an apoptosis rate of 16.4%, which was significantly higher than that of HeLa cells(6.2%, P<0.01). Compared with HeLa cells, Caspase-3 expression level in HeLa/Smac was improved significantly (P<0.01), while its activity was 3.42 times as much as that of HeLa cells (P<0.01). Conclusion: Stable transfection of extrinsic Smac gene and its over-expression in cervical cancer cell line could significantly enhance cellular caspase-3 expression and activity, ameliorate apoptosis-inducing effects of radiation on cancer cells, which would be a novel strategy to improve radiotherapeutic effects for cervical cancer. (authors)

11

Curcumin-mediated decrease in the expression of nucleolar organizer regions in cervical cancer (HeLa) cells.  

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Curcumin, the major yellow-orange pigment of turmeric derived from the rhizome of Curcuma longa, is a highly pleiotropic molecule with the potential to modulate inflammation, oxidative stress, cell survival, cell secretion, homeostasis and proliferation. Curcumin, at relatively high concentrations, was repeatedly reported to be a potent inducer of apoptosis in cancer cells and thus considered a promising anticancer agent. In the present paper, the effects of low concentrations of curcumin on human cervical cancer (HeLa) cells were studied. We found curcumin-mediated decrease in the cell number and viability, and increase in apoptotic events and superoxide level. In contrast to previously shown curcumin cytotoxicity toward different cervical cancer lines, we observed toxic effects when even as low as 1?M concentration of curcumin was used. Curcumin was not genotoxic to HeLa cells. Because argyrophilic nucleolar protein (AgNOR protein) expression is elevated in malignant cells compared to normal cells reflecting the rapidity of cancer cell proliferation, we evaluated curcumin-associated changes in size (area) and number of silver deposits. We showed curcumin-induced decrease in AgNOR protein pools, which may be mediated by global DNA hypermethylation observed after low concentration curcumin treatment. In summary, we have shown for the first time that curcumin at low micromolar range may be effective against HeLa cells, which may have implications for curcumin-based treatment of cervical cancer in humans. PMID:25308441

Lewinska, Anna; Adamczyk, Jagoda; Pajak, Justyna; Stoklosa, Sylwia; Kubis, Barbara; Pastuszek, Paulina; Slota, Ewa; Wnuk, Maciej

2014-09-01

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Effects of HMGB1 Expression Suppressed by siRNA on Cell Cycle and Proliferation of Human Cervical Cancer Cell Line HeLa  

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OBJECTIVE In this study, RNA interference was used to evaluate the effects of HMGB1 expression on cell cycle and proliferation of the human cervical cancer cell line HeLa. METHODS We had previously constructed and screened effective eukaryotic expression vectors carrying PGCsi3.0-1/HMGB1 siRNA and PGCsi3.0-3/HMGB1 siRNA, then the vectors were transfected into HeLa cells. The expression of HMGB1 before and after transfection in HeLa cells were detected by RT-PCR and Western blot. The c...

Qiu, Yuan-yuan; Wang, Hui-yu; Hao, Quan

2010-01-01

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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

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Serum ferritin in patients with cancer: determination with antibodies to HeLa cell and spleen ferritin  

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Some malignant tissues and cell lines contain acidic isoferritins and it has been suggested that the assay of such isoferritins in serum may be of value in the diagnosis of malignancy. This paper describes a radioimmunoassay for acidic ferritin purified from HeLa cells. Examination of purified heart, kidney, liver and spleen ferritin showed that the assay was highly specific for acidic isoferritins. Ferritin concentrations have been measured with antibodies to HeLa cell and spleen ferritin in extracts of normal and tumour tissue. Although the tumours contained more HeLa type ferritin than the corresponding normal tissue the HeLa/spleen type ferritin ratio was low. HeLa-type ferritin concentrations have been compared with values obtained with anti-spleen ferritin in over 1000 sera from normal subjects and patients with cancer and leukaemia. HeLa-type ferritin was not detected (<2 ?g/l) in most normal sera. Concentrations of up to 53 ?g/l were found in sera from patients with malignant disease but the HeLa/spleen type ferritin ratio was always very low. There appears to be little application for antibodies to HeLa cell or heart ferritin in the diagnosis or monitoring of cancer. (Auth.)

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Pathway of programmed cell death in HeLa cells induced by polymeric anti-cancer drugs.  

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Synthesis of anticancer polymeric materials plus their biological applications is one of the most charming and active research areas in biological functional materials. However, the predominant mechanisms for controlling cancer cell viability are not yet clear. In this work, cell culture polymeric materials co-immobilized with death signal proteins interferon-? (IFN-?)/tumor necrosis factor-? (TNF-?) on the surface were prepared by photochemical method to develop an anticancer polymeric drug model. Various characterizations on the microstructures and compositions, including the Fourier transform infrared spectroscopy, UV absorption spectroscopy, fluorescence measurement, atomic force microscopy, and electron spectroscopy for chemical analysis, were performed. For addressing the biological applications, we investigated systematically the death pathways of HeLa cells attached onto the drug model by means of a series of cell-biology techniques. It was demonstrated that the IFN-? plus TNF-? co-immobilized on the polymeric material surface exhibited more notable inhibitive effects than the free IFN-? plus TNF-?, and the induced HeLa cells were mainly along apoptosis-like PCD with the translocation of EndoG from the cytoplasm to the nucleus. These findings indicate that the polymeric drugs with the co-immobilized IFN-? plus TNF-? may offer significant potentials for therapeutic manipulation of human cervical cancer. PMID:21320725

Guan, Yan-Qing; Li, Zhibin; Chen, Jiamei; Tao, Huimin; Wang, Wenwen; Zheng, Zhe; Li, Ling; Liu, Jun-Ming

2011-05-01

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In vitro and in vivo anti-cancer activity of formononetin on human cervical cancer cell line HeLa.  

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Worldwide, cervical cancer (CC) is the third most common malignancy in women, and it remains a leading cause of cancer-related death of women. Genomic studies indicate that phosphoinositide 3-kinase (PI3K)/AKT signaling is one of the most frequently deregulated pathways in several human cancers, including CC. This signaling pathway has an important role in cancer cell proliferation, survival, motility, and metabolism, and therefore could be an attractive therapeutic target. In a previous study, we used a sensitive and high-speed homogeneous assay for the detection of kinase activity and for screening of PI3K/AKT signaling inhibitors in a high-throughput screening (HTS) format and then obtain formononetin, as an O-methylated isoflavone existed in a number of plants and herbs like Astragalus membranaceus. We showed that formononetin inhibited the phosphorylation of AKT and induced the apoptosis of CC cell line HeLa in a dose-dependent manner. Furthermore, formononetin suppressed xenograft tumor growth in nude mice. Our results indicated that formononetin may be used as an anti-cancer drug for cervical cancer in the future. PMID:24272199

Jin, Yue-mei; Xu, Tian-min; Zhao, Yan-hui; Wang, Yi-chao; Cui, Man-hua

2014-03-01

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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)

18

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

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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±2.15 µg/mL. Furthermore, treatment of HeLa cells with ICD-85 at concentrations of 1.6×10 and 2.6×10 µg/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.

Zare-Mirakabadi, Abbas; Sarzaeem, Ali

2012-01-01

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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)

20

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)

 
 
 
 
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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

22

Effect of coating material on uptake of indocyanine green-loaded nanocapsules by HeLa cervical cancer cells  

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Fluorescent molecular probes offer a potential for early cancer detection. Indocyanine green (ICG) is an FDAapproved near-infrared (NIR) fluorescent dye used in ophthalmic angiography and assessment of cardiac and hepatic functions. However, clinical applications of ICG remain very limited due to its rapid clearance from vascular circulation, unstable optical properties, non-specific interactions with plasma proteins, and inability for localized targeting. To overcome these limitations, we have encapsulated ICG within nanoconstructs composed of poly(allylamine) hydrochloride and disodium hydrogen phosphate salt. To understand the effects of coating materials on the cellular uptake of the nanocapsules, we have measured the uptake of ICG-loaded nanocapsules (ICG-NCs) with various coating materials by HeLa cancerous cervical epithelial cells in-vitro. Results of this study provide important information for the choice of appropriate coating materials that will result in maximal uptake of ICG-NCs in optical and phototherapy of cancerous tissue.

Jung, Bongsu; Lomeli, Eulieses; Anvari, Bahman

2010-02-01

23

Role of PI3K/Akt/COX-2 pathway in the radiation resistance of human cervical cancer HeLa cells  

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Objective: To explore the role of PI3K/Akt/COX-2 pathway in the radiation resistance of human cervical cancer HeLa cells. Methods: The HeLa cells were cultured in vitro. Using Celecoxib in Hela cells or associated with LY294002 for 24 h, the cells were radiated by different doses of X-ray. The cell survival ratio was detected by clone formation. To calculate Dq , D0, SF2, SER, the cell survival curve was draw by one-hit multitarget model. The expression of pAkt, Akt, COX-2, Bad and pBad were detected by Western blot. The mRNA expression of COX-2 and Bad was detected by RT-PCR. Results: The Dq, D0, SF2 of Celecoxib or associated with LY294002 group was lower than those in radiation group. The pathway PI3K/Akt/COX-2 was activated by irradiation. Celecoxib inhibited the activation of PI3K/Akt/COX-2 for multitarget. Conclusions: The activation of PI3K/Akt/COX-2 signal transduction resulted in resistance of radiosensitization in HeLa cell. It was indicated that inhibiting PI3K/Akt/COX-2 pathway could improve the radiosensitivity of human cervical cancer HeLa cells. (authors)

24

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

25

Effects of Tatariside G Isolated from Fagopyrum tataricum Roots on Apoptosis in Human Cervical Cancer HeLa Cells  

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Full Text Available Cervical cancer is the second most common female carcinoma. Current therapies are often unsatisfactory, especially for advanced stage patients. The aim of this study was to explore the effects of tatariside G (TG on apoptosis in human cervical cancer HeLa cells and the possible mechanism of action involved. An MTT assay was employed to evaluate cell viability. Hoechst 33258 staining and flow cytometry (FCM assays were used to detect cell apoptosis. The protein expression of phosphorylated JNK, P38, ERK and Akt and cleaved caspase-3 and caspase-9 was evaluated by western blot analysis. Additionally, the mRNA expression of caspase-3 and caspase-9 was measured by fluorescent quantitative reverse transcription-PCR (FQ-RT-PCR. TG notably inhibited cell viability, enhanced the percentage of apoptotic cells, facilitated the phosphorylation of p38 MAPK and JNK proteins and caspase-3 and caspase-9 cracking, downregulated the phosphorylation level of Akt, and increased the loss of MMP and the mRNA expression of caspase-3 and caspase-9. TG-induced apoptosis is associated with activation of the mitochondrial death pathway. TG may be an effective candidate for chemotherapy against cervical cancer.

Yuan Li

2014-07-01

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Effects of tatariside G isolated from Fagopyrum tataricum roots on apoptosis in human cervical cancer HeLa cells.  

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Cervical cancer is the second most common female carcinoma. Current therapies are often unsatisfactory, especially for advanced stage patients. The aim of this study was to explore the effects of tatariside G (TG) on apoptosis in human cervical cancer HeLa cells and the possible mechanism of action involved. An MTT assay was employed to evaluate cell viability. Hoechst 33258 staining and flow cytometry (FCM) assays were used to detect cell apoptosis. The protein expression of phosphorylated JNK, P38, ERK and Akt and cleaved caspase-3 and caspase-9 was evaluated by western blot analysis. Additionally, the mRNA expression of caspase-3 and caspase-9 was measured by fluorescent quantitative reverse transcription-PCR (FQ-RT-PCR). TG notably inhibited cell viability, enhanced the percentage of apoptotic cells, facilitated the phosphorylation of p38 MAPK and JNK proteins and caspase-3 and caspase-9 cracking, downregulated the phosphorylation level of Akt, and increased the loss of MMP and the mRNA expression of caspase-3 and caspase-9. TG-induced apoptosis is associated with activation of the mitochondrial death pathway. TG may be an effective candidate for chemotherapy against cervical cancer. PMID:25076146

Li, Yuan; Wang, Su-Juan; Xia, Wei; Rahman, Khalid; Zhang, Yan; Peng, Hao; Zhang, Hong; Qin, Lu-Ping

2014-01-01

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Inotodiol inhabits proliferation and induces apoptosis through modulating expression of cyclinE, p27, bcl-2, and bax in human cervical cancer HeLa cells.  

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Inonotus obliquus is a medicinal mushroom that has been used as an effective agent to treat various diseases such as diabetes, tuberculosis and cancer. Inotodiol, an included triterpenoid shows significant anti-tumor effect. However, the mechanisms have not been well documented. In this study, we aimed to explore the effect of inotodiol on proliferation and apoptosis in human cervical cancer HeLa cells and investigated the underlying molecular mechanisms. HeLa cells were treated with different concentrations of inotodiol. The MTT assay was used to evaluate cell proliferating ability, flow cytometry (FCM) was employed for cell cycle analysis and cell apoptosis, while expression of cyclinE, p27, bcl-2 and bax was detected by immunocytochemistry. Proliferation of HeLa cells was inhibited by inotodiolin a dose-dependent manner at 24h (r=0.9999, pInonotus obliquus inhibited the proliferation of HeLa cells and induced apoptosis in vitro. The mechanisms may be related to promoting apoptosis through increasing the expression of bax and cutting bcl-2 and affecting the cell cycle by down-regulation the expression of cyclin E and up-regulation of p27. The results further indicate the potential value of inotodiol for treatment of human cervical cancer. PMID:24815470

Zhao, Li-Wei; Zhong, Xiu-Hong; Yang, Shu-Yan; Zhang, Yi-Zhong; Yang, Ning-Jiang

2014-01-01

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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-?B activity. Furthermore, expression of NF-?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-?B activity, suggesting that TOPK might be a potential molecular target for successful cancer therapy using TRAIL.

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Induction of apoptotic effects of antiproliferative protein from the seeds of Borreria hispida on lung cancer (A549) and cervical cancer (HeLa) cell lines.  

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A 35 KDa protein referred to as F3 was purified from the seeds of Borreria hispida by precipitation with 80% ammonium sulphate and gel filtration on Sephadex G-100 column. RP-HPLC analysis of protein fraction (F3) on an analytical C-18 column produced a single peak, detected at 220?nm. F3 showed an apparent molecular weight of 35?KDa by SDS PAGE and MALDI-TOF-MS analyses. Peptide mass fingerprinting analysis of F3 showed the closest homology with the sequence of 1-aminocyclopropane-1-carboxylate deaminase of Pyrococcus horikoshii. The protein (F3) exhibited significant cytotoxic activity against lung (A549) and cervical (HeLa) cancer cells in a dose-dependent manner at concentrations ranging from 10?µg to 1000?µg/mL, as revealed by the MTT assay. Cell cycle analysis revealed the increased growth of sub-G0 population in both cell lines exposed to a concentration of 1000?µg/mL of protein fraction F3 as examined from flow cytometry. This is the first report of a protein from the seeds of Borreria hispida with antiproliferative and apoptotic activity in lung (A549) and cervical (HeLa) cancer cells. PMID:24605320

Rupachandra, S; Sarada, D V L

2014-01-01

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Transportation of Berberine into HepG2, HeLa and SY5Y Cells: A Correlation to Its Anti-Cancer Effect  

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The anti-cancer activities of berberine (BBR) have been reported extensively in various cancer cell lines. However, the minimal inhibitory concentrations of BBR varied greatly among different cell lines and very few studies have been devoted to elucidate this aspect. In this study, we employed three cancer cell lines, HepG2, HeLa and SY5Y, to compare the transportation and distribution of BBR. HPLC results demonstrated that BBR was capable of penetrating all the cell lines whereas the cumulative concentrations were significantly different. HepG2 cells accumulated higher level of BBR for longer duration than the other two cell lines. Molecular docking studies revealed the BBR binding site on P-glycoprotein 1 (P-gp). In addition, we elucidated that BBR regulated P-gp at both mRNA and protein levels. BBR induced the transcription and translation of P-gp in HeLa and SY5Y cells, whereas BBR inhibited P-gp expression in HepG2 cells. Further study showed that BBR regulates P-gp expression depending on different mechanisms (or affected by different factors) in different cell lines. To summarize, our study has revealed several mechanistic aspects of BBR regulation on P-gp in different cancer cell lines and might shed some useful insights into the use of BBR in the anti-cancer drug development. PMID:25402492

Pang, Yu-Nong; Liang, Yin-Wen; Feng, Tian-Shi; Zhao, Shuang; Wu, Hao; Chai, Yu-Shuang; Lei, Fan; Ding, Yi; Xing, Dong-Ming; Du, Li-Jun

2014-01-01

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

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argeting 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

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DYTOGENETIC ANALYSIS OF HELA AND CHANG CELLS  

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

N.Lzadian

1982-08-01

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Novel mixed ligand di-n-butyltin(IV) complexes derived from acylpyrazolones and fluorinated benzoic acids: synthesis, characterization, cytotoxicity and the induction of apoptosis in Hela cancer cells.  

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Twenty one novel mixed ligand di-n-butyltin(IV) complexes [(n)Bu2SnAL] (A = substituted 4-acyl-5-pyrazolone, and L = fluorinated benzoic acid) were prepared by condensation of di-n-butyltin(IV) oxide with HL and HA in 1:1:1 molar ratio in refluxing methanol. All of the complexes were characterized by elemental analyses, IR, NMR ((1)H, (13)C, (119)Sn) and in four cases by X-ray diffraction. Cytotoxicity of the compounds was studied against two human cancer cell lines (KB and Hela) by means of the MTT assay compared to cisplatin, featuring IC?? values in the low micromolar range. Hela cancer cell apoptosis-induced by 2 was examined by flow cytometry analysis, and preliminary results showed that 2 at concentrations of more than 1.0 ?M can induce apoptosis. PMID:24583378

Zhao, Bin; Shang, Xianmei; Xu, Ling; Zhang, Wendian; Xiang, Guangya

2014-04-01

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Effects of HMGB1 Expression Suppressed by siRNA on Cell Cycle and Proliferation of Human Cervical Cancer Cell Line HeLa  

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Full Text Available OBJECTIVE In this study, RNA interference was used to evaluate the effects of HMGB1 expression on cell cycle and proliferation of the human cervical cancer cell line HeLa. METHODS We had previously constructed and screened effective eukaryotic expression vectors carrying PGCsi3.0-1/HMGB1 siRNA and PGCsi3.0-3/HMGB1 siRNA, then the vectors were transfected into HeLa cells. The expression of HMGB1 before and after transfection in HeLa cells were detected by RT-PCR and Western blot. The cell viability and proliferating activity was tested by Trypan blue dye test and MTT, and the cell cycle was determined by ? ow cytometry. RESULTS The introduction of PGCsi3.0-1/HMGB1 siRNA and PGCsi3.0-3/HMGB1 siRNA inhibited the expression of HMGB1mRNA and protein efficiently and specifically, there was a sifnificant difference between the siRNA groups and the control groups (P < 0.05. The proliferation speed of PGCsi3.0-1 group and PGC si3.0-3 group were obviously slower than those of PGCsi3.0-Neg group and non-transfected group. Flow cytometry showed that the content of DNA in G2 phase in PGCsi3.0-1 group and PGCsi3.0-3 group were obviously more than those in PGCsi3.0-Neg group and non-transfected group, but the content in S phase was less (P < 0.01. The progression of cell cycle was arrested from G2 to S phase. CONCLUSION PGCsi3.0-1/HMGB1 siRNA and PGCsi3.0-3/HMGB1 siRNA could specially suppress the expression of HMGB1 gene, inhibit the proliferation speed of HeLa cells effectively, and arrest the progression of cell cycle from G2 to S phase. RNAi provides a new approach to the bio-therapy of cervical cancer.

Yuan-yuan QIU

2010-04-01

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[6]-Gingerol induces caspase 3 dependent apoptosis and autophagy in cancer cells: drug-DNA interaction and expression of certain signal genes in HeLa cells.  

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[6]-Gingerol, a pharmacologically important bioactive component of ginger, has been reported to have anti-hyperglycemic, anti-cancer and anti-oxidative properties, but mechanisms through which these are achieved are largely unclear. The present study focuses on apoptosis and autophagy, two key events of anti-cancer activity, in HeLa cells treated with [6]-gingerol. The treated cells showed several morphological changes, including externalization of phosphatidyl serine, degradation of DNA and increase in TUNEL positivity. Furthermore, there was depolarization of mitochondrial membrane potential, providing evidence of mitochondria mediated apoptosis. The expression of caspase 3 and PARP was increased in cells exposed to [6]-gingerol. Circular dichroism study for testing drug-DNA interaction with both calf thymus and nuclear DNA as target revealed that the drug had potential to bind with the nuclear DNA and induce conformational changes of DNA. The over-expression of NFk?, AKT and Bcl2 genes in cancer cells was down-regulated by [6]-gingerol treatment. On the other hand the expression levels of TNF?, Bax and cytochrome c were enhanced in [6]-gingerol treated cells. Thus, overall results suggest that [6]-gingerol has potential to bind with DNA and induce cell death by autophagy and caspase 3 mediated apoptosis. PMID:22939973

Chakraborty, Debrup; Bishayee, Kausik; Ghosh, Samrat; Biswas, Raktim; Mandal, Sushil Kumar; Khuda-Bukhsh, Anisur Rahman

2012-11-01

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Influences of ionizing radiation on apoptosis and cell cycle of HelaS3 cell lines  

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Changes of apoptosis and cell cycle progression of HeLaS3 cells following irradiation with different doses of X rays were observed. The percentages of apoptotic cells were examined by flow cytometry (FCM) after the cells had been stained with prospidium iodine (PI) and Hoechst 33342 (HO), and the cell cycle was detected by FCM stained with only PI. It was found that the percentages of apoptotic cells and the cells in S phase and G2/M phase increased after irradiation with 0.5-4.0 Gy X rays. So irradiation may induce apoptosis and cell cycle block for HelaS3 cell lines

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Evaluating the gene-expression profiles of HeLa cancer cells treated with activated and nonactivated poly(amidoamine) dendrimers, and their DNA complexes.  

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Using dendrimers in cancer therapy as nonviral vectors for gene delivery seems promising. The biological performance of a dendrimer-based gene delivery system depends heavily on its molecular architecture. The transfection activity of dendrimers is significantly improved by processes activated in the heat degradation treatment of solvolysis. However, very little is known about the molecular mechanisms that dendrimers produce in cancer cells. We studied the changes in global gene-expression profiles in human cervical cancer HeLa cells exposed to nonactivated and activated poly(amidoamine) (PAMAM) dendrimers, alone or in complexes with plasmid DNA (dendriplexes). Real-time quantitative reverse transcriptase-polymerase chain reaction was used to confirm four regulated genes (PHF5A, ARNTL2, CHD4, and P2RX7) affected by activated dendrimers and dendriplexes. Activated and nonactivated dendrimers and dendriplexes alike induced multiple gene expression changes, some of which overlapped with their dendriplexes. Dendrimer activation improved transfection efficiency and induced additional gene expression changes in HeLa cells. Dendrimers and dendriplexes principally affect genes with the molecular functions of nucleic acid binding and transcription activity, metal-ion binding, enzyme activity, receptor activity, and protein binding. Our findings provide a deeper insight into the changes in gene expression patterns caused by the molecular structure of PAMAM dendrimers for gene-based cancer therapy. PMID:20394435

Kuo, Jung-hua Steven; Liou, Meng-jie; Chiu, Hsueh-chen

2010-06-01

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The aqueous extract of Ficus religiosa induces cell cycle arrest in human cervical cancer cell lines SiHa (HPV-16 Positive) and apoptosis in HeLa (HPV-18 positive).  

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

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

2013-01-01

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Irradiation And Papillomavirus E2 Proteins On Hela Cells  

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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

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Potentiation of Apoptin-induced apoptosis by Cecropin B-like antibacterial peptide ABPs1 in human HeLa cervical cancer cell lines is associated with membrane pore formation and caspase-3 activation.  

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Apoptin, a chicken anemia virus-encoded protein, induces apoptosis in chicken or human tumor cells, localizing in their nuclei as opposed to the cytoplasm of non-transformed cells. The present study was undertaken to investigate whether ABPs1 could potentiate apoptininduced apoptosis in HeLa cells. ABPs1 and the apoptin genes were successfully cloned into pIRES2-EGFP expression vector and expressed in HeLa cells. We report that ABPs1 augments apoptin cell growth inhibition in a concentration- and time-dependent manner. The DAPI staining and scanning electron microscopy observations revealed apoptotic bodies and plasma membrane pores, which were attributed to apoptin and ABPs1, respectively. Further, ABPs1 in combination with apoptin was found to increase the expression of Bax and to decrease the expression of survivin compared with either agent alone or the control. The apoptotic rate of HeLa cells treated with ABPs1 and apoptin in combination for 48 h was 53.95%. The two-gene combination increased the caspase-3 activity of HeLa cells. Taken together, our study suggests that ABPs1 combined with apoptin significantly inhibits HeLa cell proliferation, and induces cell apoptosis through membrane defects, up-regulation of Bax expression, down-regulation of survivin expression, and activation of the caspase-3 pathway. Thus, the combination of ABPs1 and apoptin could serve as a means to develop novel gene therapeutic agents against human cervical cancer. PMID:24633228

Birame, Basse Mame; Jigui, Wang; Fuxian, Yu; Jiazeng, Sun; Zhili, Li; Weiquan, Liu

2014-06-28

 
 
 
 
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Effect of 17-AAG on radio-sensitivity of HeLa and V79 cells  

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In order to investigate the radio-sensitizing effect of 17-AAG, an inhibitor of Heat Shock Protein 90, on human Uterine Cervix Cancer HeLa and V79 cells, Clonogenic assay was used to observe the cell survival rate. The results show that 17-AAG can decrease obviously (p0.05). This indicates that 17-AAG may enhance the radio-sensitivity of the HeLa cell line and has no effect on the V79 cell line. (authors)

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Effect of rhodoxanthin from Potamogeton crispus L. on cell apoptosis in Hela cells.  

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Carotenoid, a natural functional pigment, is known to have anti-carcinogenic activity. To verify the anti-cancer effects of rhodoxanthin which is a kind of carotenoids, we investigated the effects of rhodoxanthin from Potamogeton crispus L. on the proliferation rate, cell cycle distribution, apoptosis and the change in mitochondrial membrane potential in Hela cell line. The effects of rhodoxanthin were also tested on the concentration of Ca(2+) in cells. Rhodoxanthin inhibited cell proliferation in Hela cells in a dose and time-dependent manner. Rhodoxanthin induced an accumulation of cells in the S phase of the cell cycle, reduced the mitochondria transmembrane potential and increased the concentration of intracellular Ca(2+). In summary, our results suggested that rhodoxanthin-induced apoptosis in Hela cells occurred via these pathways. PMID:16919415

Ren, Dandan; Peng, Guanghua; Huang, Hongxia; Wang, Haibin; Zhang, Shenghua

2006-12-01

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Regulation of radiosensitivity by HDAC inhibitor trichostatin A in the human cervical carcinoma cell line Hela.  

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Histone deacetylase (HDAC) inhibitors play an important role in inducing growth arrest, differentiation, and/or apoptosis in cancer cells. Given their ability to disrupt critical biological processes in cancer cells, these agents are emerging as potential therapeutics for cancer. Recently, it has been identified that HDAC inhibitors can also efficiently enhance the radiation sensitivity of cells, both in vitro and in vivo. In this study, we investigated whether the potent HDAC inhibitor, Trichostatin A, modulates the radiation sensitivity of the human cervical carcinoma cell line Hela under hypoxic conditions. We concluded that TSA could significantly inhibit the proliferation of Hela cells in a dose-and time-dependent manner under normoxic and hypoxic conditions. Hypoxia resulted in the cervical carcinoma Hela cells resistant to TSA. The findings from clonogenic survival assays indicate that incubation with TSA for 24 hours prior to irradiation enhances the radiation sensitivity of Hela cells under hypoxic conditions. More generally, we found Hela cells under hypoxic conditions treated with TSA could significantly down-regulate the expressions of HIF-1alpha and VEGF proteins. Taken together, our results demonstrated that TSA acts as a powerful radiosensitizer in Hela cells under hypoxic conditions probably by down-regulated expression of HIF-1alpha and VEGF proteins. PMID:22873101

Yu, J; Mi, J; Wang, Y; Wang, A; Tian, X

2012-01-01

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Naringin inhibits growth and induces apoptosis by a mechanism dependent on reduced activation of NF??B/COX?2?caspase-1 pathway in HeLa cervical cancer cells.  

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Naringin (NRG), a bioflavonoid found in citrus fruit extracts, has been pharmacologically evaluated as a potential anticancer agent. This study confirmed a novel mechanism of the anticancer effects of NRG in the human cervical cancer HeLa cell line (HeLa cells). Exposure of HeLa cells to NRG resulted in growth inhibition, as evidenced by a decrease in cell viability. In addition, NRG treatment induced apoptosis, as indicated by the increased apoptotic percentage and the cleaved caspase-3 expression. Importantly, exposure of the cells to NRG attenuated the expression levels of phosphorylated (p) nuclear factor ?B (NF-?B) p65 subunit, cyclooxygenase-2 (COX-2) and cysteinyl aspartate proteinase-1 (caspase-1). Treatment with PDTC (an inhibitor of NF-?B) or NS-398 (an inhibitor of COX-2) or SC-3069 (an inhibitor of caspase-1) markedly induced growth inhibition and apoptosis. Treatment with PDTC or NS-398 also reduced caspase-1 expression. Interestingly, PDTC treatment blocked the expression of COX-2 and NS-398 reduced the p-NF-?B p65 expression. Taken together, this study provides novel evidence that NRG induces growth inhibition and apoptosis by inhibiting the NF-?B/COX-2-caspase-1 pathway and that a positive interaction between NF-?B and COX-2 pathway contributes to the growth and antiapoptotic effect in HeLa cells. PMID:25174821

Zeng, Lan; Zhen, Yulan; Chen, Yiming; Zou, Lin; Zhang, Ying; Hu, Fen; Feng, Jianqiang; Shen, Jianhua; Wei, Bing

2014-11-01

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Effect of Quercetin on radio-sensitivity of HeLa cells  

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In order to investigate the mechanism of Quercetin on radio-sensitivity of human Uterine Cervix Cancer HeLa cells, HeLa cells were cultured in different concentrations of Quercetin and different doses of irradiation. The clonogenic assay was used to observe the cell survival rate. The repair of DNA double-strand breaks and effect of Quercetin combination of radiation on the cell cycle were detected by flow cytometry. The results show that the radio-sensitivity of Quercetin on HeLa cells was obvious and the unrepaired DSBs after irradiation increased, but did not decrease G2/M cell cycle arrest. From this it can be inferred that the effect on HeLa cell radio-sensitivity may be related to the inhibition of the repair of DNA double-strand breaks induced by Quercetin, but it dose not reveal a significant relation with the cell cycle and G2/M arrest. (authors)

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

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

Wei ZHANG

2013-02-01

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

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

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  

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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  

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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.

José María Eloy, Contreras-Ortiz; Juan Carlos, Vázquez-Chagoyán; José Simón, Martínez-Castañeda; José Guillermo, Estrada-Franco; José Esteban, Aparicio-Burgos; Jorge, Acosta-Dibarrat; Alberto, Barbabosa-Pliego.

2013-09-02

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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

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Suberoyl bishydroxamic acid-induced apoptosis in HeLa cells via ROS-independent, GSH-dependent manner.  

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Suberoyl bishydroxamic acid (SBHA) is a HDAC inhibitor that can regulate many biological functions including apoptosis and proliferation in various cancer cells. Here, we evaluated the effect of SBHA on the growth of HeLa cervical cancer cells in relation to apoptosis, reactive oxygen species (ROS) and glutathione (GSH) levels. Dose-dependent inhibition of cell growth was observed in HeLa cells with an IC50 of approximately 15 ?M at 72 h. SBHA also induced apoptosis in HeLa cells, as evidenced by sub-G1 cells, annexin V-FITC staining cells, activations of caspase 3 and 8, and the loss of mitochondrial membrane potential (??m). In addition, all of the tested caspase inhibitors rescued some cells from SBHA-induced HeLa cell death. SBHA increased ROS levels including O2(•-) and induced GSH depletion in HeLa cells. Generally, caspase inhibitors did not affect ROS levels in SBHA-treated HeLa cells, but they significantly prevented GSH depletion in these cells. Furthermore, while the well-known antioxidants, N-acetyl cysteine and vitamin C, did not affect cell death, ROS level or GSH depletion in SBHA-treated HeLa cells, L-buthionine sulfoximine, a GSH synthesis inhibitor, enhanced cell death and GSH depletion in these cells. In conclusion, SBHA inhibits the growth of HeLa cervical cancer cells via caspase-dependent apoptosis, and the inhibition is independent of ROS level changes, but dependent on GSH level changes. PMID:23269626

You, Bo Ra; Park, Woo Hyun

2013-05-01

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Radiosensitization of nitroindazole derivatives on HeLa cells  

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Objective: To investigate the cytotoxicity and radiosensitization of 5-nitroindazole-3-formyliminodiacetic acid on HeLa cells. Methods: HeLa cells in exponential growth phase were incubated in culture media with different doses and the survival rate was determined by MTT assay. The survival rate of cells receiving radiation combined with different doses of medicine was compared with that of the control.Results: The cytotoxicity of S-nitroindazole-3-formyliminodiacetic acid on HeLa cells was very low. The drug had hypoxia radiosensitizing effect on HeLa cells. At doses of 0, 6, 12, 24, 48 and 96 ?g/ml under hypoxia, the survival rate were 0.91 , 0.87, 0.84, 0.81, 0.76 and 0.60, respectively. At the dosage of 48 and 96 ?g/ml, the survival rate were 0.85 and 0.73 under oxygenous). Conclusions: 5-Nitroindazole-3-formyliminodiacetic acid has low cytotoxicity and rediosensitizing effect on HeLa cells. (authors)

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

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Trichostatin A (TSA) as a HDAC inhibitor can regulate many biological properties including apoptosis and cell proliferation in various cancer cells. Here, we evaluated the effect of TSA on the growth and death of HeLa cervical cancer cells in relation to reactive oxygen species (ROS) and glutathione (GSH) levels. Dose- and time-dependent growth inhibition was observed in HeLa cells with an IC50 of approximately 20 nM at 72 h. This agent also induced apoptotic cell death, as evidenced by annexin V-FITC staining cells, caspase-3 activation and the loss of mitochondrial membrane potential (MMP; ??m). In addition, the administration of Bcl-2 siRNA intensified TSA-induced HeLa cell death. All of the tested caspase inhibitors significantly rescued some cells from TSA-induced HeLa cell death. TSA increased O2•- level and induced GSH depletion in HeLa cells. Caspase inhibitors significantly attenuated O2•- level and GSH depletion in TSA-treated HeLa cells. In addition, N-acetyl cysteine (NAC; a well known antioxidant) significantly prevented cell death and GSH depletion in TSA-treated HeLa cells via decreasing O2•- level. In conclusion, TSA inhibited the growth of HeLa cells via Bcl-2-mediated apoptosis, which was closely related to O2•- and GSH content levels. PMID:23165748

You, Bo Ra; Park, Woo Hyun

2013-01-01

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Autophagy facilitates Salmonella replication in HeLa cells.  

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Autophagy is a process whereby a double-membrane structure (autophagosome) engulfs unnecessary cytosolic proteins, organelles, and invading pathogens and delivers them to the lysosome for degradation. We examined the fate of cytosolic Salmonella targeted by autophagy and found that autophagy-targeted Salmonella present in the cytosol of HeLa cells correlates with intracellular bacterial replication. Real-time analyses revealed that a subset of cytosolic Salmonella extensively associates with autophagy components p62 and/or LC3 and replicates quickly, whereas intravacuolar Salmonella shows no or very limited association with p62 or LC3 and replicates much more slowly. Replication of cytosolic Salmonella in HeLa cells is significantly decreased when autophagy components are depleted. Eventually, hyperreplication of cytosolic Salmonella potentiates cell detachment, facilitating the dissemination of Salmonella to neighboring cells. We propose that Salmonella benefits from autophagy for its cytosolic replication in HeLa cells. IMPORTANCE As a host defense system, autophagy is known to target a population of Salmonella for degradation and hence restricting Salmonella replication. In contrast to this concept, a recent report showed that knockdown of Rab1, a GTPase required for autophagy of Salmonella, decreases Salmonella replication in HeLa cells. Here, we have reexamined the fate of Salmonella targeted by autophagy by various cell biology-based assays. We found that the association of autophagy components with cytosolic Salmonella increases shortly after initiation of intracellular bacterial replication. Furthermore, through a live-cell imaging method, a subset of cytosolic Salmonella was found to be extensively associated with autophagy components p62 and/or LC3, and they replicated quickly. Most importantly, depletion of autophagy components significantly reduced the replication of cytosolic Salmonella in HeLa cells. Hence, in contrast to previous reports, we propose that autophagy facilitates Salmonella replication in the cytosol of HeLa cells. PMID:24618251

Yu, Hong B; Croxen, Matthew A; Marchiando, Amanda M; Ferreira, Rosana B R; Cadwell, Ken; Foster, Leonard J; Finlay, B Brett

2014-01-01

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

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

Alcindo A. Dos Santos

2013-01-01

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ANTICANCER AND CYTOTOXIC POTENTIAL OF TRITICUM AESTIVUM EXTRACT ON HELA CELL LINE  

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The objective of the study was to analyze the anticancer property of the leaves of Triticum aestivum on HeLa cells. The Indian medicinal plant Triticum aestivum that is used in traditional medicine for cancer and non cancerous diseases was collected. The crude methanolic extract was prepared by using standard protocols. The antiproliferative effect the methanolic extract was evaluated in vitro by employing MTT assay. The potency of each plant extract concentration was calculated in terms of p...

Patel Janki B.; Patel Piyush M.

2013-01-01

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Expression of LGI1 Impairs Proliferation and Survival of HeLa Cells  

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

Nadia Gabellini

2009-01-01

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Biocompatibility of Porous Spherical Calcium Carbonate Microparticles on Hela Cells  

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

Xu Yang

2012-03-01

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Growth regulation of HeLa cells by 1060 nm photons  

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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)

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

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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; Semwal, Manoj K.; Sarma, Asitikantha; Avasthi, Devesh Kumar

2013-04-01

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

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

Kathirvel, Poonkodi; Ravi, Subban

2012-01-01

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Genetic transformation of HeLa cells by Agrobacterium  

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Agrobacterium tumefaciens is a soil phytopathogen that elicits neoplastic growths on the host plant species. In nature, however, Agrobacterium also may encounter organisms belonging to other kingdoms such as insects and animals that feed on the infected plants. Can Agrobacterium, then, also infect animal cells? Here, we report that Agrobacterium attaches to and genetically transforms several types of human cells. In stably transformed HeLa cells, the integration ev...

Kunik, Talya; Tzfira, Tzvi; Kapulnik, Yoram; Gafni, Yedidya; Dingwall, Colin; Citovsky, Vitaly

2001-01-01

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Antiproliferative effects of Tanaceti partheni, Hypericum perforatum and propolis on HeLa cells  

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Full Text Available Tanaceti partheni, Hypericum perforatum and propolis have been widely used for centuries and are well-documented medicinal plants and natural product. In this study, we investigated the antiproliferative effects of water extracts of ethanolic dry extracts of two different medicinal plants (Tanaceti partheni and Hypericum perforatum and propolis on HeLa cells. The Tanaceti partheni extract exhibited mild cytotoxic activity. The IC50 was 153.71 ?g/mL. The extract of Hypericum perforatum did not show active cytotoxic activity against HeLa cells (IC50 >200 ?g/mL. Regarding the antiproliferative effects of Hypericum perforatum, our results are not in correlation with the results of other authors, probably because different Hypericum species and different human cancer cell lines were used. The extract of propolis did not show active cytotoxic activity against HeLa cells (IC50 = 1.08 ± 0.01 mg/mL. The weak antiproliferative effect of propolis on HeLa cells is either due to the use of a low concentration of propolis extracted in weakly polar solvents, or the use of propolis collected in the autumn. [Projekat Ministarstva nauke Republike Srbije, br. 34021 i br. 175011

Ceni?-Miloševi? Desanka

2014-01-01

64

Phosphatidylinositol anchor of HeLa cell alkaline phosphatase  

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

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ANTICANCER AND CYTOTOXIC POTENTIAL OF TRITICUM AESTIVUM EXTRACT ON HELA CELL LINE  

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Full Text Available The objective of the study was to analyze the anticancer property of the leaves of Triticum aestivum on HeLa cells. The Indian medicinal plant Triticum aestivum that is used in traditional medicine for cancer and non cancerous diseases was collected. The crude methanolic extract was prepared by using standard protocols. The antiproliferative effect the methanolic extract was evaluated in vitro by employing MTT assay. The potency of each plant extract concentration was calculated in terms of percent decrease in viable HeLa cells as compared to the control value. The extract showed dose dependent antitumor activity. The MTT assay showed an anti proliferative activity (IC50 at 156 ?g/ml of crude extract.

Patel Janki B.

2013-01-01

66

Energy metabolism in hela and walker-256 cells  

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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)

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AMP-activated protein kinase is required for cell survival and growth in HeLa-S3 cells in vivo.  

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Activation of the AMP-dependent protein kinase (AMPK) is linked to cancer cell survival in a variety of cancer cell lines, particularly under conditions of stress. As a potent activator of AMPK, metformin has become a hot topic of discussion for its effect on cancer cell. Here, we report that AMPK activated by metformin promotes HeLa-S3 cell survival and growth in vivo. Our results show that metformin inhibited cell proliferation in MCF-7 cells, but not in LKB1-deficient HeLa-S3 cells. Re-expression of LKB-1 in HeLa-S3 cells restored the growth inhibitory effect of metformin, indicating a requirement for LKB-1 in metformin-induced growth inhibition. Moreover, AMPK activation exerted a protective effect in HeLa-S3 cells by relieving ER stress, modulating ER Ca(2+) storage, and finally contributing to cellular adaptation and resistance to apoptosis. Our findings identify a link between AMPK activation and cell survival in HeLa-S3 cells, which demonstrates a beneficial effect of AMPK activated by metformin in cancer cell, and suggests a discrete re-evaluation on the role of metformin/AMPK activation on tumor cell growth, proliferation, and on clinical application in cancer therapy. PMID:24916949

Song, Xuhong; Huang, Dongyang; Liu, Yanmin; Pan, Xiaokang; Zhang, Jing; Liang, Bin

2014-06-01

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

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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.

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

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

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

Zahra Tayarani-Najarani

2012-04-01

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

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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.

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

2012-04-01

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Microcell-mediated transfer of chromosome 4 into HeLa cells suppresses telomerase activity.  

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Telomerase activity can be detected in most human cancers and immortal cell lines. In contrast, the lack of telomerase activity in normal diploid fibroblasts has been correlated with progressive reduction of telomere lengths to critically short sizes followed by the cessation of cell division and the onset of senescence. Several investigators have provided evidence for the localization of a telomerase suppressor gene on chromosome 3. The aim of our study was to determine whether other chromosomes are involved in telomerase repression. Beside human chromosome 3 (serving as positive control), chromosomes 4, 6, and 11 were introduced into HeLa cells via microcell-mediated chromosome transfer. Telomerase activity from different hybrid cell lysates was determined at an early time point after fusion using a Telomerase ELISA kit. Strong repression of telomerase activity was only found in a subset of HeLa hybrids in which chromosome 3 or chromosome 4 had been introduced. Telomerase suppression induced by chromosome 3 or 4 transfer was paralleled by a high frequency (30% or 43%, respectively) of a senescent-like phenotype. Chromosomes 6 and 11, the functional loss of which is also implicated in cervical cancer, had no effect. These results indicate that normal human chromosomes 3 and 4 carry a gene or genes that suppress telomerase activity and induce cellular senescence in HeLa cells. PMID:11319808

Backsch, C; Wagenbach, N; Nonn, M; Leistritz, S; Stanbridge, E; Schneider, A; Dürst, M

2001-06-01

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

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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

73

Three-dimensional printing of Hela cells for cervical tumor model in vitro.  

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Advances in three-dimensional (3D) printing have enabled the direct assembly of cells and extracellular matrix materials to form in vitro cellular models for 3D biology, the study of disease pathogenesis and new drug discovery. In this study, we report a method of 3D printing for Hela cells and gelatin/alginate/fibrinogen hydrogels to construct in vitro cervical tumor models. Cell proliferation, matrix metalloproteinase (MMP) protein expression and chemoresistance were measured in the printed 3D cervical tumor models and compared with conventional 2D planar culture models. Over 90% cell viability was observed using the defined printing process. Comparisons of 3D and 2D results revealed that Hela cells showed a higher proliferation rate in the printed 3D environment and tended to form cellular spheroids, but formed monolayer cell sheets in 2D culture. Hela cells in 3D printed models also showed higher MMP protein expression and higher chemoresistance than those in 2D culture. These new biological characteristics from the printed 3D tumor models in vitro as well as the novel 3D cell printing technology may help the evolution of 3D cancer study. PMID:24722236

Zhao, Yu; Yao, Rui; Ouyang, Liliang; Ding, Hongxu; Zhang, Ting; Zhang, Kaitai; Cheng, Shujun; Sun, Wei

2014-09-01

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

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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

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

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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)

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Toona Sinensis and Moschus Decoction Induced Cell Cycle Arrest in Human Cervical Carcinoma HeLa Cells.  

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Toona sinensis and Moschus are two herb materials used in traditional Chinese medicine, most commonly for their various biological activities. In this study, we investigated the inhibitory effect of three decoctions from Toona sinensis, Moschus, and Toona sinensis and Moschus in combination on cell growth in several normal and cancer cell lines by cell viability assay. The results showed that the combined decoction exhibited the strongest anticancer effects, compared to two single decoctions. The observations indicated that the combined decoction did not induce cell apoptosis and autophagy in HeLa cells by fluorescence microscopy. Flow cytometry analysis revealed that the combined decoction arrested HeLa cell cycle progression in S-phase. After the decoction incubation, among 41 cell cycle related genes, eight were reduced, while five were increased in mRNA levels by real-time PCR assay. Western blotting showed that there were no apparent changes of protein levels of Cyclin E1, while P27 expression significantly declined and the levels of CDC7 and CDK7 obviously increased. The data suggest that the RB pathway is partially responsible for the decoction-induced S-phase cell cycle arrest in HeLa cells. Therefore, the combined decoction may have therapeutic potential as an anticancer formula for certain cancers. PMID:24511319

Zhen, Hong; Zhang, Yifei; Fang, Zhijia; Huang, Zhiwei; You, Chongge; Shi, Ping

2014-01-01

77

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)

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Stable tRNA precursors in HeLa cells.  

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Two tRNA precursors were isolated from 32P-labeled or unlabeled HeLa cells by two dimensional polyacrylamide gel electrophoresis, and were sequenced. These were the precursors of tRNAMet and tRNALeu, and both contained four extra nucleotides including 5'-triphosphates at their 5'-end and nine extra nucleotides including oligo U at their 3'-end. These RNAs are the first naturally occurring tRNA precursors from higher eukaryotes whose sequences have been determined. In these molecules, several ...

Harada, F.; Matsubara, M.; Kato, N.

1984-01-01

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Molekulêre kruiskommunikasie tussen apoptose en outofagie geïnduseer deur ‘n 2-metoksi?stradiol analoog (C19 in HeLa selle Molecular crosstalk between apoptosis and autophagy induced by a 2-methoxyestradiol analogue (C19 in HeLa cells.  

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Full Text Available Servikale kanker is gerapporteer deur die Wêreld Gesondheids Organisasie as die mees algemene tipe kanker wat vroue in armer sosio-ekonomiese lande affekteer.Using a novel synthesised sulphamoylated 2-methoxyestradiol analogue, C19, two types of cell death, namely apoptosis and autophagy, were demonstrated in vitro when cervical cancer HeLa cells were exposed to this compound.

A. E. Theron

2012-03-01

80

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

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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 ca

 
 
 
 
81

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

82

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

83

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

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MicroRNA-21 promotes cell proliferation and down-regulates the expression of programmed cell death 4 (PDCD4) in HeLa cervical carcinoma cells  

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MicroRNAs are involved in cancer-related processes. The microRNA-21(miR-21) has been identified as the only miRNA over-expressed in a wide variety of cancers, including cervical cancer. However, the function of miR-21 is unknown in cervical carcinomas. In this study, we found that the inhibition of miR-21 in HeLa cervical cancer cells caused profound suppression of cell proliferation, and up-regulated the expression of the tumor suppressor gene PDCD4. We also provide direct evidence that PDCD4-3'UTR is a functional target of miR-21 and that the 18 bp putative target site can function as the sole regulatory element in HeLa cells. These results suggest that miR-21 may play an oncogenic role in the cellular processes of cervical cancer and may serve as a target for effective therapies.

85

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

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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

87

Comprehensive and definitive molecular cytogenetic characterization of HeLa cells by spectral karyotyping.  

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We revisited the cytogenetic alterations of the cervical adenocarcinoma cell line HeLa through the use of spectral karyotyping (SKY), comparative genomic hybridization (CGH), and fluorescence in situ hybridization (FISH). SKY analysis unequivocally characterized all abnormal chromosomes. Chromosomal breakpoints were primarily assigned by simultaneous assessment of SKY painted chromosomes and inverted 4,6-diamidino2-phenylindole banding from the same cell. Twenty clonally abnormal chromosomes were found. Comparison with previously reported HeLa G-banding karyotypes revealed a remarkably stable cytogenetic constitution because 18 of 20 markers that were found were present before. The classification of 12 markers was refined in this study. Our assignment of the remaining six markers was consistent with those described in the literature. The CGH map of chromosomal copy number gains and losses strikingly matched the SKY results and was, in a few instances, decisive for assigning breakpoints. The combined use of molecular cytogenetic methods SKY, CGH, and FISH with site-specific probes, in addition to inverted 4,6-diamidino-2-phenylindole or conventional G-banding analysis, provides the means to fully assess the genomic abnormalities in cancer cells. Human papillomaviruses (HPVs) are frequently integrated into the cellular DNA in cervical cancers. We mapped by FISH five HPV18 integration sites: three on normal chromosomes 8 at 8q24 and two on derivative chromosomes, der(5)t(5;22;8)(qll;q11q13;q24) and der(22)t(8; 22)(q24;q13), which have chromosome 8q24 material. An 8q24 copy number increase was detected by CGH. Dual-color FISH with a c-MYC probe mapping to 8q24 revealed colocalization with HPV18 at all integration sites, indicating that dispersion and amplification of the c-MYC gene sequences occurred after and was most likely triggered by the viral insertion at a single integration site. Numerical and structural chromosomal aberrations identified by SKY, genomic imbalances detected by CGH, as well as FISH localization of HPV18 integration at the c-MYC locus in HeLa cells are common and representative for advanced stage cervical cell carcinomas. The HeLa genome has been remarkably stable after years of continuous cultivation; therefore, the genetic alterations detected may have been present in the primary tumor and reflect events that are relevant to the development of cervical cancer. PMID:9892199

Macville, M; Schröck, E; Padilla-Nash, H; Keck, C; Ghadimi, B M; Zimonjic, D; Popescu, N; Ried, T

1999-01-01

88

Evaluation of the effects of paederus beetle extract and gamma irradiation on HeLa cells  

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Objective(s): Cervical cancer is a malignancy that is the second most common cause of death from cancer in women throughout the world. Paederus beetle (Paederus fuscipes) extract (PBE), contains bioactive compounds such as pederine which has cytotoxic properties and blocks DNA and protein synthesis at very low concentrations. In this investigation we tried to determine the effects co-treatment with PBE and gamma irradiation on HeLa cells. Materials and Methods: The viability of the cells was measured by two methods: MTT and Colony assay. Results: We found that supplementing gamma irradiation therapy with PBE does not increase cell death and it might even interfere with its cytotoxicty at the concentrations below 0.1 ng/ml and the viability for irradiation vs irradiation + PBE was 37%: 60%. Conclusion: This finding might be due to radioprotective effects of the very low doses of PBE against gamma radiation. PMID:24904724

Samani, Fariba; Monfared, Ali Shabestani; Zabihi, Ebrahim; Khafri, Soraya; Karimi, Maesoumeh; Akhavan Niaki, Haleh

2014-01-01

89

Evaluation of cytotoxicity of Moringa oleifera Lam. callus and leaf extracts on Hela cells  

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Background: There are considerable attempts worldwide on herbal and traditional compounds to validate their use as anti-cancer drugs. Plants from Moringaceae family including Moringa oleifera possess several activities such as antitumor effect on tumor cell lines. In this study we sought to determine if callus and leaf extracts of M. oleifera possess any cytotoxicity. Materials and Methods: Ethanol-water (70-30) extracts of callus and leaf of M. oleifera were prepared by maceration method. The amount of phenolic compounds of the extracts was determined by Folin Ciocalteu method. The cytotoxicity of the extracts against Hela tumor cells was carried out using MTT assay. Briefly, cells were seeded in microplates and different concentrations of the extract were added. Cells were incubated for 48 h and 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. Cytotoxicity was considered when more than 50% reduction on cell survival was observed. Results: Callus and leaf extracts of M. oleifera significantly decreased the viability of Hela cells in a concentration-dependent manner. However, leaf extract of M. oleifera were more potent than that of callus extract. Conclusion: As the content of phenolic compounds of leaf extract was higher than that of callus extract, it can be concluded that phenolic compounds are involved in the cytotoxicity of M. oleifera. PMID:25337524

Jafarain, Abbas; Asghari, Gholamreza; Ghassami, Erfaneh

2014-01-01

90

Effects of 3-AB on PARP expression of Hela cells and apoptosis and cell cycle progression of Hela cells after X-rays irradiation  

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Objective: To study the changes of apoptosis and cell cycle progression of Hela cells after the poly (ADP- ribose) polymerase (PARP) was inhibited by its inhibitor 3-aminobenzamid (3-AB) and the mechanisms of PARP interaction with Hela cells damaged by irradiation. Methods: Hela cell line was used. Flow cytometry (FCM) was used to examine the PARP expression of control and 3 AB groups at 0, 2, 4, 8, 12 h alter administration with 5 mmol·L-1 3-AB. The percentage of apoptotic cells and cell cycle progression ol control, irradiation, 3-AB plus irradiation groups were measured with FCM at 2, 8, 12, 24 h after exposure to 2 Gy irradiation following administration with 5 mmol·L-1 3-AB. Results: The percentage of Hela cells with positive expression of PARP protein decreased after administration with 3-AB and there was significant difference between 3-AB plus irradiation group and control group (P2 cells in the 3-AB plus irradiation group were lower than those in the irradiation group (P2 arrest induced by irradiation. (authors)

91

Cytotoxic effect and radiation enhancement of artemisinin in uterine cervical carcinoma cell line HeLa  

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Objective: To investigate cytotoxic and radiosensitizing effect of Artemisinin on cervical carcinoma cell line HeLa. Methods: In order to measure the optimized effective time, cytotoxic effect of Artemisinin on HeLa cell line was investigated with MTT assay. The radiosensitization effect of different doses and different treatment duration of Artemisinin on HeLa cell line were evaluated by MTT test, the SER is 1.17 and radiosensitizing effect was measured with multi-target single hit model through SER of HeLa cell. Cell cycles in different groups were calculated by flow cytometry. Results: The 50% inhibition concentration of Artemisinin interacted with HeLa cells for 24 h is 600.19 nmol/ml, and for 48 h is 160.71 nmol/ml. The HeLa cells'surival ratio is 93.51%, 91.87%, and 87.28% after adding Atemisinin of 110.69 nmol/ml and 1 Gy radiation exposure. There are three groups: the chemotherapy only group, the radiotherapy only group and the combination group. The result of the cell cycles showed that cells in G2/M period decreased in the combination group. Conclusion: Artemisinin has radiosensitization effect on cervical carcinoma HeLa cells, whichshows dose and time dependent. Artemisinin can inhibit the G2/M block by ionizing radiation. (authors)

92

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

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

Joanna Jakubowicz-Gil

2012-10-01

93

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

94

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

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

Alshatwi Ali A

2009-11-01

95

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

96

Proteasome inhibitor lactacystin enhances cisplatin cytotoxicity by increasing endoplasmic reticulum stress?associated apoptosis in HeLa cells.  

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Cisplatin is commonly used as a therapeutic agent, despite its known adverse side effects and the occurrence of drug resistance. The development of novel methods for combination therapy with cisplatin is required in order to circumvent these limitations of cisplatin alone. The proteasome inhibitor lactacystin (LAC) has been indicated to produce anti?tumor effects, and has previously been used as an antitumor agent in cancer treatment research; however, its effects in combination with cisplatin treatment are unknown. In the current study, the effects of LAC in combination with cisplatin treatment were investigated in HeLa human cervical cancer (HCC) cells. The results demonstrated that cisplatin treatment inhibited cell growth and induced cell apoptosis. HeLa cell exposure to cisplatin induced endoplasmic reticulum (ER) stress?associated apoptosis, and LAC treatment increased levels of cell apoptosis and the activation of caspase?3. Specifically, LAC treatment increased the cisplatin?induced expression of PDI, GRP78, CHOP, cleaved caspase?4 and cleaved caspase?3. Together, these data indicate that LAC is able to enhance cisplatin cytotoxicity by increasing ER stress?associated apoptosis in HeLa cells. PMID:25323748

Xu, Ye; Li, Di; Zeng, Linchuan; Wang, Chunyan; Zhang, Lili; Wang, Yan; Yu, Yang; Liu, Shibing; Li, Zhixin

2015-01-01

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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

98

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

99

Apigenin inhibits HeLa sphere?forming cells through inactivation of casein kinase 2?.  

Science.gov (United States)

The protein kinase casein kinase 2 (CK2) has been implicated in stem cell maintenance and its aberrant activation has been demonstrated in several types of cancer, including cervical cancer. In the present study, it was demonstrated that the sphere?forming cells (SFCs) of HeLa cell lines exhibited self?renewal capacity, indicating that they possessed the properties of cervical cancer stem?like cells. HeLa?derived SFCs exhibited a higher level of CK2? protein, compared with the parental cells. Apigenin, a dietary flavonoid, led to a dose?dependent inhibition of the self?renewal capacity and the protein expression of CK2? in HeLa?derived SFCs. Furthermore, forced overexpression of CK2? resulted in a decrease in the inhibition of CK2? expression and the self?renewal capacity induced by apigenin in HeLa?derived SFCs. These results suggested that apigenin inhibits the self?renewal capacity of HeLa?derived SFCs through downregulation of CK2? expression. PMID:25334018

Liu, Jie; Cao, Xiao-Cheng; Xiao, Qiao; Quan, Mei-Fang

2015-01-01

100

Cell-cycle-dependent variations in the FTIR spectroscopy of HeLa cells treated with trichostatin A.  

Science.gov (United States)

It is quite complex to evaluate the mechanism of action for antitumor drugs on cancer cells. Studies have pointed out that there is an unique advantage of Fourier transform infrared spectrum to obtain a fingerprint of all molecules present in the cells when cancer cells were exposed to anti-cancer drugs. Trichostatin A (TSA) is a most potent reversible inhibitor of mammalian histone deacetylases. It can inhibit cancer cell growth in vitro and in vivo. In the present study, HeLa cells were exposed to 0, 50, 100, 200, 300 and 400 nmol x L(-1) TSA, and FTIR spectra were applied to evaluate the effect of TSA on cancer cells. Results show that there is some significant relationship between the changes in FTIR absorption and cell cycle arresting. On the other hand, this investigation shows that the concentration of TSA had to be more than 200 nmol x L(-1) in order to ensure A1080 cm(-1)/A1540 cm(-1) > or = 1 for inhibiting cell proliferation. PMID:22007388

Zhang, Feng-Qiu; Qi, Jian; Yang, Zhan-Guo

2011-08-01

 
 
 
 
101

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

102

Optimizing A Lipocomplex-Based Gene Transfer Method into HeLa Cell Line  

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Full Text Available One of the most significant steps in gene expression studies is transferring genes into cell cultures. Despite there are different methods for gene delivery such as viral and non-viral producers, some cationic lipid reagents have recently developed to transfect into mammalian cell lines. The main aim of this study was optimizing and improving lipocomplex based transient transfection procedures into HeLa cell line which is being used widely as a typical cell in biological studies.This study was an experimental research. In this work, pCMV: ?-Gal DNA plasmid was used as a reporter DNA for determining the rate of gene transfection into HeLa cells. To accomplish the highest gene delivery into HeLa cells, optimizing experiments were carried out in different volumes of FuGENE-HD, LipofectamineTM2000 and X-tremeGENE. Also, we investigated tranasfection efficiency in presence of various cell densities of HeLa cells. Then, transfection efficiency and cell toxicity were measured by beta gal staining and trypan blue methods, respectively.Using FuGENE-HD in volume of 4?l along with 105 HeLa cells, transfection efficiency was higher (43.66 ± 1.52% in comparison with the cationic lipids lipofectamineTM2000 and X-tremeGENE. In addition, the rate of cell toxicity in presence of FuGENE-HD was less than 5%.In summary, the cationic lipid FuGENE-HD indicates a suitable potential to transfer DNA into HeLa cells and it can be an efficient reagent for gene delivery for HeLa cells in vitro. Moreover, it is worth designing and optimizing gene transfer experiments for other cell lines with FuGENE-HD due to its low toxicity and high efficiency.

Alimohammad Asgharian

2013-01-01

103

Rheological properties of mammalian cell culture suspensions: Hybridoma and HeLa cell lines.  

Science.gov (United States)

Data on viscous (eta') and elastic (eta'') components of the complex viscosity versus oscillatory angular frequency (0.01 to 4.0 rad/s) with increasing strains were obtained for hybridoma cell (62'D3) and HeLa cell (S3) suspensions in PBS at 0.9 (mL/mL) cell volume fraction using a Weissenberg rheogoniometer equipped with two parallel plate geometry at ambient temperature. Both cell suspensions exhibited shear thinning behavior. From the measured viscoelastic properties, the yield stress was calculated. Hybridoma cell suspension (15 microm as the mean diameter of cells) showed the yield stress at 550 dyne/cm(2) that was 1.8 times higher than the value of HeLa cell suspension (22 microm mean diameter) as measured at the oscillatory angular frequency, 4.0 rad/s. The apparent viscosities of HeLa cell suspension at four concentrations and varying steady shear rate were also determined using the Brookfield rotational viscometer. The yield stress to steady shear test was about 130 dyne/cm(2) for HeLa cell suspension at 0.9 (mL/mL) cell volume fraction. The apparent viscosity was in the range about 1 approximately 1000 Poise depending on the cell concentration and shear rate applied. A modified semiempirical Mooney equation, eta = eta(0) exp[K gamma(.)(-beta)phi(c)(1 - K'' sigmaphi(c) /D)] was derived based on the cell concentration, the cell morphology, and the steady shear rate. The beta, shear rate index, was estimated as 0.159 in the range of shear rate, 0.16 to 22.1 s(-1), for the cell volume fractions from 0.6 to 0.9 (mL/mL). In this study, the methods of determining the shear sensitivity and the viscous and the elastic components of mammalian cell suspensions are described under the steady shear field. PMID:18609617

Shi, Y; Ryu, D D; Ballica, R

1993-03-25

104

Modification of radiation response in HeLa cells by misonidazole  

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The hypoxic radiosensitizer misonidazole decreased survival in dense cultures of HeLa cells irradiated with gamma rays in a non-dose modifying fashion. Survival curves of treated hypoxic cells displayed a much larger extrapolation number than untreated cells. In oxygenated randomly dividing cells, drug treatment had an effect opposite to that in hypoxic cells, increasing survival. In cultures initiated from mitotic cells and irradiated soon afterwards, a smaller sensitization under hypoxia and no increase in survival of oxygenated cells was observed. It was concluded that metabolic as well as radiochemical events take place in misonidazole-treated and then irradiated HeLa cells which modify survival. (author)

105

Effect of hyperthermia and radiation on the cell cycle progression of HeLa cells  

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The effect of hyperthermia and irradiation on cytokinetics was studied using exponentially growing HeLa cells. To determine the effect of heat and/or radiation on the cell cycle progression, the changes in the DNA distribution of the cell population after time intervals after treatment were studied. The cellular DNA content of the cell population was measured by flow cytometry. The results obtained were as follows: 1. Compared with the control, the cellular DNA content distribution of HeLa cells treated with 430C for 20 min and 60 min showed cell accumulation in S and G2M phases 8 hours after treatment. 2. Hyperthermic treatment at 450C for 20 min caused cells to accumulate in S phase in the first 4 hours and G2M phase after 8 to 14.5 hours, whereas heat treatment at 450C for 60 min caused cells to accumulate in G2M phase after 24 hours. 3. Irradiation of exponentially growing cells induced a block in the progress from G2M to G1 phase. 4. Dose survival curves of HeLa cells with and without postirradiation thermal treatment (430C, 60 min) showed significant enhancement of radiosensitivity by hyperthermia. 5. The sequential treatment, i.e. 5 Gy irradiation followed immediately by heat treatment at 430C for 60 min, caused more cells to accumulate in G2M phase after 24 and 48 hours, as compared with 5 Gy irradiation alone. (author)

106

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

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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)

107

Ionizing radiation regulated expression of Lipofectamin-mediated p16 gene in HeLa cells  

International Nuclear Information System (INIS)

Objective: To investigate the expression of Egr-p16 and its role in tumor cell inhibition in HeLa cells. Methods: HeLa cells were transfected with p16 by recombined plasmids with Lipofectamin induced by ?-ray irradiation with different doses. RT-PCR was used to detect p16 mRNA level in the dose-effect and time-course aspects. Cell proliferation was detected by cell count and cell cycle changes were detected by FCM. Results: The results showed that p16 mRNA level was higher than the control after 0.5-8.0 Gy irradiation. There were peaks after 2-4 Gy irradiation, and p16 mRNA level increased at 2-4 h after 2 Gy irradiation. There was a significant increase at 4 h. After the stably transfected HeLa cells were irradiated, the transfer of Egr-p16 combined with ?-ray irradiation could inhibit the growth of HeLa cells. The percentage of G0/G1 showed a dose-dependant decrease, but the percentage of S showed a dose-dependant increase. The percentage of G2/M showed a markedly arrest after 2 Gy irradiation, and gradually reduced after 5 and 10 Gy irradiation, but it was still higher than the control group. Conclusion: ?-rays can induce the recombined plasmid to express at mRNA level in HeLa cells and there was a significant change in cell proliferation

108

Experimental study on the killing effect of 125I seeds to Hela cells  

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Objective: To investigate the rules of apoptosis and proliferation of Hela cells in vitro irradiated with 125I seeds and the killing effect to tumor cells. Methods: The proliferation reaction and morphological change of Hela cells with low energy ? ray 125I irradiation in different dose and duration were observed. Results: The apoptosis segments of Hela cells irradiated with 125I appeared was on stage II b. However, significant increase in the cell apoptosis ratio related to the dose accumulation and apoptosis reached peak level at a total dose of 5 Gy. meanwhile, the results of the cell clones generally correlated negatively with apoptosis and demonstrated a significant reduction. A reverve dose rate effect was also found. Conclusion: The therapeutic effect of 131I seeds irradiation probably may be the inhibition of cell proliferation in early stage, and apoptosis of large amount of tumor cells in later stage. (authors)

109

Cell Culture on Nanopillar Sheet: Study of HeLa Cells on Nanopillar Sheet  

Science.gov (United States)

This is the first report of the successful culturing of HeLa cells on nanopillar sheets—a new type of cell culture dish—in a different way from that on flat petri dishes. Nanopillar sheets were fabricated with a high-aspect ratio structure with a diameter of 80-1000 nm and a height of 1-3 ?m using nanoprint technology. Nanopillar structure with 500 nm diameter and 1 ?m height enabled easy subculture of the cells from the sheets without the conventional trypsinization method. Moreover, the HeLa cells divided and proliferated on the sheets in the same way as on the flat surfaces with different manner of adhesion.

Nomura, Shinobu; Kojima, Hiroko; Ohyabu, Yoshimi; Kuwabara, Kosuke; Miyauchi, Akihiro; Uemura, Toshimasa

2005-09-01

110

Characterization of the association of radiolabeled bleomycin A2 with HeLa cells  

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The association of [3H]bleomycin A2 and Cu(II):[3H]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 [3H]bleomycin A2 or Cu(II):[3H]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

111

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

112

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

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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

113

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

114

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

Science.gov (United States)

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

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

2009-01-01

115

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

116

Targeting Diamond Nanoparticles into Folate-Receptor Expressing HeLa Cells  

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We have studied binding of synthesized folic acid-diamond nanoparticle conjugates to proliferatively active HeLa cells. In order to determine the binding of the complex to the cells, we used spectral luminescence methods and microscopy, which let us visualize localization of the conjugate in the cellular system in vitro. We show that the conjugate under study binds to folate-receptor expressing HeLa cells. We have established a determining role for the folate receptor in binding of the conjugate to the cells. Our studies suggest that is it possible to use the conjugate as a targeted nanoplatform for targeted delivery of diagnostic and therapeutic agents to tumor cells.

Lapina, V. A.; Vorobey, A. V.; Pavich, T. A.; Opitz, J.

2013-07-01

117

Reciprocal relationship between cytosolic NADH and ENOX2 inhibition triggers sphingolipid-induced apoptosis in HeLa cells.  

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ENOX2 (tNOX), a tumor-associated cell surface ubiquinol (NADH) oxidase, functions as an alternative terminal oxidase for plasma membrane electron transport. Ubiquitous in all cancer cell lines studied thus far, ENOX2 expression correlates with the abnormal growth and division associated with the malignant phenotype. ENOX2 has been proposed as the cellular target for various quinone site inhibitors that demonstrate anticancer activity such as the green tea constituent epigallocatechin-3-gallate (EGCg) and the isoflavone phenoxodiol (PXD). Here we present a possible mechanism that explains how these substances result in apoptosis in cancer cells by ENOX2-mediated alterations of cytosolic amounts of NAD(+) and NADH. When ENOX2 is inhibited, plasma membrane electron transport is diminished, and cytosolic NADH accumulates. We show in HeLa cells that NADH levels modulate the activities of two pivotal enzymes of sphingolipid metabolism: sphingosine kinase 1 (SK1) and neutral sphingomyelinase (nSMase). Their respective products sphingosine 1-phosphate (S1P) and ceramide (Cer) are key determinants of cell fate. S1P promotes cell survival and Cer promotes apoptosis. Using plasma membranes isolated from cervical adenocarcinoma (HeLa) cells as well as purified proteins of both bacterial and human origin, we demonstrate that NADH inhibits SK1 and stimulates nSMase, while NAD(+) inhibits nSMase and has no effect on SK1. Additionally, intact HeLa cells treated with ENOX2 inhibitors exhibit an increase in Cer and a decrease in S1P. Treatments that stimulate cytosolic NADH production potentiate the antiproliferative effects of ENOX2 inhibitors while those that attenuate NADH production or stimulate plasma membrane electron transport confer a survival advantage. PMID:20518072

De Luca, Thomas; Morré, Dorothy M; Morré, D James

2010-08-15

118

Construction of a model cell line for the assay of MDR1 (multi drug resistance gene-1) substrates/inhibitors using HeLa cells.  

Science.gov (United States)

Cancer cells often become resistant to chemotherapy, and induction of the ABC transporter Multi-drug Resistance gene-1 (MDR1) is a major cause. We established a tool for high-throughput screening of substrates and inhibitors of MDR1, using transformed HeLa cells that over-express MDR1. The cDNA for human MDR1 was subcloned into the eukaryotic expression vector pBK-CMV to produce an MDR1 expression vector, pBK-CMV/MDR1. HeLa cells were transfected with pBK-CMV/MDR1 or the empty vector pBK-CMV. Transfection of the vector sequence for MDR1 and its expression were evaluated by genomic PCR and western blotting, respectively. The efficiency of the MDR1 transporter for pumping a substrate out of the transformed cells was evaluated using rhodamine123 (R-123), a mitochondrial dye that is also an MDR1 substrate. After treatment of the MDR1-expressing HeLa cells with MDR1 substrate vinblastin or inhibitors cyclosporin A and verapamil, the amount of R-123 retained in the cells was increased to 2 to 2.3 times the level in untreated MDR1-expressing HeLa cells. The transfection of empty pBK-CMV had no effect on the R-123 retention in HeLa cells, regardless of drug treatment. In conclusion, we have established a model human carcinoma cell line that expresses functional MDR1 and can be used to screen for substrates and inhibitors of MDR1. PMID:19530439

Kugawa, F; Suzuki, T; Miyata, M; Tomono, K; Tamanoi, F

2009-05-01

119

Curcumin targeting the thioredoxin system elevates oxidative stress in HeLa cells  

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The thioredoxin system, composed of thioredoxin reductase (TrxR), thioredoxin (Trx), and NADPH, is ubiquitous in all cells and involved in many redox-dependent signaling pathways. Curcumin, a naturally occurring pigment that gives a specific yellow color in curry food, is consumed in normal diet up to 100 mg per day. This molecule has also been used in traditional medicine for the treatment of a variety of diseases. Curcumin has numerous biological functions, and many of these functions are related to induction of oxidative stress. However, how curcumin elicits oxidative stress in cells is unclear. Our previous work has demonstrated the way by which curcumin interacts with recombinant TrxR1 and alters the antioxidant enzyme into a reactive oxygen species (ROS) generator in vitro. Herein we reported that curcumin can target the cytosolic/nuclear thioredoxin system to eventually elevate oxidative stress in HeLa cells. Curcumin-modified TrxR1 dose-dependently and quantitatively transfers electrons from NADPH to oxygen with the production of ROS. Also, curcumin can drastically down-regulate Trx1 protein level as well as its enzyme activity in HeLa cells, which in turn remarkably decreases intracellular free thiols, shifting the intracellular redox balance to a more oxidative state, and subsequently induces DNA oxidative damage. Furthermore, curcumin-pretreated HeLa cells are more sensitive to oxidative stress. Knockdown of TrxR1 sensitizes HeLa cells to curcumin cytotoxicity, highlighting the physiological significance of targeting TrxR1 by curcumin. Taken together, our data disclose a previously unrecognized prooxidant mechanism of curcumin in cells, and provide a deep insight in understanding how curcumin works in vivo. -- Highlights: ? Curcumin induces oxidative stress by targeting the thioredoxin system. ? Curcumin-modified TrxR quantitatively oxidizes NADPH to generate ROS. ? Knockdown of TrxR1 augments curcumin's cytotoxicity in HeLa cells. ? Curcumin sensitizes HeLa cells to oxidative stress.

Cai, Wenqing; Zhang, Baoxin; Duan, Dongzhu [State Key Laboratory of Applied Organic Chemistry, Lanzhou University, Lanzhou, Gansu 730000 (China); Wu, Jincai [College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, Gansu 730000 (China); Fang, Jianguo, E-mail: fangjg@lzu.edu.cn [State Key Laboratory of Applied Organic Chemistry, Lanzhou University, Lanzhou, Gansu 730000 (China); College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, Gansu 730000 (China)

2012-08-01

120

Curcumin targeting the thioredoxin system elevates oxidative stress in HeLa cells  

International Nuclear Information System (INIS)

The thioredoxin system, composed of thioredoxin reductase (TrxR), thioredoxin (Trx), and NADPH, is ubiquitous in all cells and involved in many redox-dependent signaling pathways. Curcumin, a naturally occurring pigment that gives a specific yellow color in curry food, is consumed in normal diet up to 100 mg per day. This molecule has also been used in traditional medicine for the treatment of a variety of diseases. Curcumin has numerous biological functions, and many of these functions are related to induction of oxidative stress. However, how curcumin elicits oxidative stress in cells is unclear. Our previous work has demonstrated the way by which curcumin interacts with recombinant TrxR1 and alters the antioxidant enzyme into a reactive oxygen species (ROS) generator in vitro. Herein we reported that curcumin can target the cytosolic/nuclear thioredoxin system to eventually elevate oxidative stress in HeLa cells. Curcumin-modified TrxR1 dose-dependently and quantitatively transfers electrons from NADPH to oxygen with the production of ROS. Also, curcumin can drastically down-regulate Trx1 protein level as well as its enzyme activity in HeLa cells, which in turn remarkably decreases intracellular free thiols, shifting the intracellular redox balance to a more oxidative state, and subsequently induces DNA oxidative damage. Furthermore, curcumin-pretreated HeLa cells are more sensitive to oxidative stress. Knockdown of TrxR1 sensitizes HeLa cells to curcumin cytotoxicity, highlighting the physiological significance of targeting TrxR1 by curcumin. Taken together, our data disclose a previously unrecognized prooxidant mechanism of curcumin in cells, and provide a deep insight in understanding how curcumin works in vivo. -- Highlights: ? Curcumin induces oxidative stress by targeting the thioredoxin system. ? Curcumin-modified TrxR quantitatively oxidizes NADPH to generate ROS. ? Knockdown of TrxR1 augments curcumin's cytotoxicity in HeLa cells. ? Curcumin sensitizes HeLa cells to oxidative stress.

 
 
 
 
121

tRNA modifying enzymes, NSUN2 and METTL1, determine sensitivity to 5-fluorouracil in HeLa cells.  

Science.gov (United States)

Nonessential tRNA modifications by methyltransferases are evolutionarily conserved and have been reported to stabilize mature tRNA molecules and prevent rapid tRNA decay (RTD). The tRNA modifying enzymes, NSUN2 and METTL1, are mammalian orthologs of yeast Trm4 and Trm8, which are required for protecting tRNA against RTD. A simultaneous overexpression of NSUN2 and METTL1 is widely observed among human cancers suggesting that targeting of both proteins provides a novel powerful strategy for cancer chemotherapy. Here, we show that combined knockdown of NSUN2 and METTL1 in HeLa cells drastically potentiate sensitivity of cells to 5-fluorouracil (5-FU) whereas heat stress of cells revealed no effects. Since NSUN2 and METTL1 are phosphorylated by Aurora-B and Akt, respectively, and their tRNA modifying activities are suppressed by phosphorylation, overexpression of constitutively dephosphorylated forms of both methyltransferases is able to suppress 5-FU sensitivity. Thus, NSUN2 and METTL1 are implicated in 5-FU sensitivity in HeLa cells. Interfering with methylation of tRNAs might provide a promising rationale to improve 5-FU chemotherapy of cancer. PMID:25233213

Okamoto, Mayumi; Fujiwara, Mamoru; Hori, Masato; Okada, Kaoru; Yazama, Futoshi; Konishi, Hiroaki; Xiao, Yegui; Qi, Guangying; Shimamoto, Fumio; Ota, Takahide; Temme, Achim; Tatsuka, Masaaki

2014-09-01

122

p53-dependent Fas expression is critical for Ginsenoside Rh2 triggered caspase-8 activation in HeLa cells.  

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We have recently reported that Ginsenoside Rh2 (G-Rh2) induces the activation of two initiator caspases, caspase-8 and caspase-9 in human cancer cells. However, the molecular mechanism of its death-inducing function remains unclear. Here we show that G-Rh2 stimulated the activation of both caspase-8 and caspase-9 simultaneously in HeLa cells. Under G-Rh2 treatment, membrane death receptors Fas and TNFR1 are remarkably upregulated. However, the induced expression of Fas but not TNFR1 was contributed to the apoptosis process. Moreover, significant increases in Fas expression and caspase-8 activity temporally coincided with an increase in p53 expression in p53-non-mutated HeLa and SK-HEP-1 cells upon G-Rh2 treatment. In contrast, Fas expression and caspase-8 activity remained constant with G-Rh2 treatment in p53-mutated SW480 and PC-3 cells. In addition, siRNA-mediated knockdown of p53 diminished G-Rh2-induced Fas expression and caspase-8 activation. These results indicated that G-Rh2-triggered extrinsic apoptosis relies on p53-mediated Fas over-expression. In the intrinsic apoptotic pathway, G-Rh2 induced strong and immediate translocation of cytosolic BAK and BAX to the mitochondria, mitochondrial cytochrome c release, and subsequent caspase-9 activation both in HeLa and in SW480 cells. p53-mediated Fas expression and subsequent downstream caspase-8 activation as well as p53-independent caspase-9 activation all contribute to the activation of the downstream effector caspase-3/-7, leading to tumor cell death. Taken together, we suggest that G-Rh2 induces cancer cell apoptosis in a multi-path manner and is therefore a promising candidate for anti-tumor drug development. PMID:24622841

Guo, Xiao-Xi; Li, Yang; Sun, Chao; Jiang, Dan; Lin, Ying-Jia; Jin, Feng-Xie; Lee, Seung-Ki; Jin, Ying-Hua

2014-03-01

123

Isolating HeLa Cell Fractions Enriched for Clathrin-Coated Vesicles.  

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HeLa cell lines can be experimentally manipulated using drugs or gene-silencing techniques such as RNA interference. Fractions enriched for clathrin-coated vesicles (CCVs) can be isolated from these cell lines and used to study the effects of these manipulations on the composition of CCVs. This protocol, originally developed in the laboratory of Margaret Robinson (Cambridge, United Kingdom), describes the preparation of a HeLa cell fraction that is enriched for a mixed population of CCVs and is suitable for analysis by mass spectroscopy, western blotting, or electron microscopy. PMID:25368312

Borner, Georg H H; Fielding, Andrew B

2014-01-01

124

Effects of red light emitting diode on apoptosis of HeLa cells and suppression of implanted HeLa cells growth in mice.  

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Low intensity irradiation of cells by laser was an effective method of biostimulation. Here, we have extended these actions to evaluate the apoptosis effects in red light emitting diode (RLED) exposure. Through morphological observation, flow cytometric analysis, intracellular calcium measurement and RT-PCR, we found that HeLa cells in 24 h RLED irradiation in in-vitro experiments would significantly affects the induction of cellular apoptosis, and morphological changes such as the loose arrangement of cells, the noticeable development of apoptotic bodies,and the accompaniment of arrested S phase and activated caspases-3,-6,-8. Moreover, intracellular calcium concentrations markedly increased 40.3 +/- 1.3% and 43.1 +/- 0.8% respectively, relative to an extracellular solution containing the Ca(2+) and Ca(2+) free unexposed group. In in-vivo tests, RLED irradiation decreased the growth of tumors on day 50 and attenuated the elevation of vascular endothelial growth factor (VEGF) expression in HeLa cell implanted BALB/c mice. Taken together, our results suggest that RLED could induce HeLa cell apoptosis and convey potential antitumor properties. PMID:19164885

Zhang, Li; Xiong, Zhe; Li, Zhengjia; Yao, Bei; Zhang, Donghua

2009-03-01

125

DNA polymerases ?, ?, and var-epsilon: Three distinct enzymes from HeLa cells  

International Nuclear Information System (INIS)

DNA polymerases ?, ?, ? have been purified and characterized from the same HeLa cell extract in order to determine their relationship by comparing them from the same cell type. The catalytic properties and the primary structures of the large subunits of the DNA polymerases as compared by partial peptide mapping with N-chlorosuccinimide are different. Likewise, the small subunit of DNA polymerase ? appears to be distinct from the large subunit of the same polymerase and from the smaller subunits of DNA polymerase ?. HeLa DNA polymerase ? is processive only when HeLa proliferating cell nuclear antigen is present, whereas DNA polymerase ? is quite processive in its absence. Inhibitor and activator spectra of DNA polymerases ?, ?, and ? also distinguish the three enzymes. These results and immunologic comparisons published elsewhere support the premise that HeLaDNA polymerases ?, ?, and ? are distinct enzymes that have common properties with yeast DNA polymerases I, HI, and II, respectively

126

Study of photodynamic, sonodynamic and antioxidative influence on HeLa cell line.  

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Photodynamic treatment (PDT) in combination with sonodynamic treatment (SDT) can be used as suitable methods to treat malignant and benign diseases or combat resistant bacteria. Both methods affect the production of reactive oxygen species (ROS). On the other hand, antioxidants are useful for cell protection against ROS. This work was aimed to study the effect of PDT and SDT treatments on the HeLa cell line using antioxidant Pronalen Sensitive Skin as a protection from free radicals in the cells. We evaluated the effect of sensitizer ClAlPcS2 using battery of in vitro methods, including MTT assay, kinetic production of ROS, mitochondrial membrane potential change, type of cell death and microscopic analysis. Ultrasound treatment was observed to increase the production of ROS, only in combination with PDT, particularly at higher concentrations of ClAlPcS2. The added antioxidant acts as protection against free radicals and has potential as a dietary supplement against aging or free radicals. The results of study suggested that ClAlPcS2 could be used as a potential photosensitizer for treatment of a specific type of cancers. PMID:24791413

Tomankova, Katerina; Kolarova, Hana; Vachutka, Jaromir; Zapletalova, Jana; Hanakova, Adela; Kaplova, Eva

2014-02-01

127

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

128

Single-walled carbon nanotube interactions with HeLa cells  

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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

129

Trypanosoma cruzi trypomastigotes induce cytoskeleton modifications during HeLa cell invasion  

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

Maria Cecília Fernandes

2011-12-01

130

Aphidicolin does not inhibit DNA repair synthesis in ultraviolet-irradiated HeLa cells  

International Nuclear Information System (INIS)

A radioautographic examination of nuclear DNA synthesis in unirradiated and u.v.-irradiated HeLa cells, in the presence and in the absence of aphidicolin, showed that aphidicolin inhibits nuclear DNA replication and has no detectable effect on DNA repair synthesis. Although the results establish that in u.v.-irradiated HeLa cells most of the DNA repair synthesis is not due to DNA polymerase ?, they do not preclude a significant role for this enzyme in DNA repair processes. (author)

131

Oropouche virus entry into HeLa cells involves clathrin and requires endosomal acidification.  

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Oropouche virus (ORO), family Bunyaviridae, is the second most frequent cause of arboviral febrile illness in Brazil. Studies were conducted to understand ORO entry in HeLa cells. Chlorpromazine inhibited early steps of ORO replication cycle, consistent with entry/uncoating. The data indicate that ORO enters HeLa cells by clathrin-coated vesicles, by a mechanism susceptible to endosomal acidification inhibitors. Transmission electron microscopy and immunofluorescence indicated that ORO associates with clathrin-coated pits and can be found in association with late endosomes in a time shorter than 1h. PMID:18840482

Santos, Rodrigo I M; Rodrigues, Alcir H; Silva, Maria Lúcia; Mortara, Renato A; Rossi, Marcos A; Jamur, Maria Célia; Oliver, Constance; Arruda, Eurico

2008-12-01

132

Present status of MA-160 cell line. Prostatic epithelium or HeLa cells.  

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This report reviews the background and the present status of the MA-160 cell line. This cell line was reported to have arisen from tissue cultures of a human, benign prostatic adenoma and consequently has been used for investigations on the prostat and on hormone-dependent cells. Recent studies using chromosome analysis by banding techniques, glucose-6-phosphate dehydrogenase (G6D) electrophoretic mobility, and a specific prostatic acid phosphatase test indicate that MA-160 is not of prostatic origin but a HeLa cell contaminant, arising as a result of intraspecies cell contamination. It is suggested that whenever spontaneous transformation is observed in vitro and established cell lines are being used in the laboratory, the cells in question be thoroughly checked with the currently available chromosomal, biochemical, and cytochemical methods in order to establish their origin and to rule out interspecies and intraspecies cell contamination. PMID:66218

Webber, M M; Horan, P K; Bouldin, T R

1977-03-01

133

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

International Nuclear Information System (INIS)

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

134

The Roles of Platelet GPIIb/IIIa and ?v?3 Integrins during HeLa Cells Adhesion, Migration, and Invasion to Monolayer Endothelium under Static and Dynamic Shear Flow  

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Full Text Available During their passage through the circulatory system, tumor cells undergo extensive interactions with various host cells including endothelial cells and platelets. Mechanisms mediating tumor cell adhesion, migration, and metastasis to vessel wall under flow condition are largely unknown. The aim of this study was to investigate the potential roles of GPIIb/IIIa and ?v?3 integrins underlying the HeLa-endothelium interaction in static and dynamic flow conditions. HeLa cell migration and invasion were studied by using Millicell cell culture insert system. The numbers of transmigrated or invaded HeLa cells significantly increased by thrombin-activated platelets and reduced by eptifibatide, a platelet inhibitor. Meanwhile, RGDWE peptides, a specific inhibitor of ?v?3 integrin, also inhibited HeLa cell transmigration. Interestingly, the presence of endothelial cells had significant effect on HeLa cell migration regardless of static or cocultured flow condition. The adhesion capability of HeLa cells to endothelial monolayer was also significantly affected by GPIIb/IIIa and ?v?3 integrins. The arrested HeLa cells increased nearly 5-fold in the presence of thrombin-activated platelets at shear stress condition (1.84?dyn/cm2 exposure for 1 hour than the control (static. Our findings showed that GPIIb/IIIa and ?v?3 integrins are important mediators in the pathology of cervical cancer and provide a molecular basis for the future therapy, and the efficient antitumor benefit should target multiple receptors on tumor cells and platelets.

Yiyao Liu

2009-01-01

135

Study of Paclitaxel-Treated HeLa Cells by Differential Electrical Impedance Flow Cytometry  

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Full Text Available This work describes the electrical investigation of paclitaxel-treated HeLa cells using a custom-made microfluidic biosensor for whole cell analysis in continuous flow. We apply the method of differential electrical impedance spectroscopy to treated HeLa cells in order to elucidate the changes in electrical properties compared with non-treated cells. We found that our microfluidic system was able to distinguish between treated and non-treated cells. Furthermore, we utilize a model for electrical impedance spectroscopy in order to perform a theoretical study to clarify our results. This study focuses on investigating the changes in the electrical properties of the cell membrane caused by the effect of paclitaxel. We observe good agreement between the model and the obtained results. This establishes the proof-of-concept for the application in cell drug therapy.

Julie Kirkegaard

2014-08-01

136

Study of Paclitaxel-Treated HeLa Cells by Differential Electrical Impedance Flow Cytometry  

DEFF Research Database (Denmark)

This work describes the electrical investigation of paclitaxel-treated HeLa cells using a custom-made microfluidic biosensor for whole cell analysis in continuous flow. We apply the method of differential electrical impedance spectroscopy to treated HeLa cells in order to elucidate the changes in electrical properties compared with non-treated cells. We found that our microfluidic system was able to distinguish between treated and non-treated cells. Furthermore, we utilize a model for electrical impedance spectroscopy in order to perform a theoretical study to clarify our results. This study focuses on investigating the changes in the electrical properties of the cell membrane caused by the effect of paclitaxel. We observe good agreement between the model and the obtained results. This establishes the proof-of-concept for the application in cell drug therapy.

Kirkegaard, Julie; Clausen, Casper Hyttel

2014-01-01

137

Cytotoxic Effects of Different Extracts and Latex of Ficus carica L. on HeLa cell Line.  

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It has been reported that latex and extracts of different species of Ficus are cytotoxic to some human cancerous cell lines. In this study, cytotoxicity of fruit and leaf extracts as well as the latex of Ficuscarica L. on HeLa cell line were evaluated. ethanolic extracts of leaves and fruits were prepared through percolation and ethyl acetate and dichloromethane extracts were prepared by reflux method. Cytotoxic effects of these extracts and latex against HeLa cell line were then examined. Briefly, He Lacells were seeded at 2 × 10(4) cells/mL in 96-well plates. After 24 h incubation at 37(°)C, the cells were treated with different concentrations of the extracts or latex. The viability of the cells was determined by the reduction of 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) from formazan following 48 h incubation and the absorbance was measured at 540 nm using an ELISA plate reader. The results indicated that the latex and different extracts of Ficus carica could reduce the viability of the He Lacells at concentrations as low as 2 µg/mL in a dose dependent manner. The approximate IC50 values of the ethanolic, ethyl acetate and dichloromethane extracts of the leaves and fruits were 10, 19, 12 µg/mL and 12, 12, 11.5 µg/mL, respectively. The IC50 for the latex was about 17 µg/mL. PMID:24250354

Khodarahmi, Ghadam Ali; Ghasemi, Nasrollah; Hassanzadeh, Farshid; Safaie, Marzieh

2011-01-01

138

The effect of aclacinon on macromolecular syntheses in HeLa cells  

International Nuclear Information System (INIS)

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.)

139

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

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

Chan Wai-Yee

2006-06-01

140

Mechanism of aminoglycoside enhancement of Staphylococcus aureus adherence to HeLa cells.  

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There is enhanced adherence of Staphylococcus aureus to HeLa cells if the organism is grown in the presence of sub-lethal concentration of aminoglycosides. In this study, we investigated the mechanism of this enhancement. Cell surface components obtained by lysosaphin digestion under hypertonic conditions were examined for binding to HeLa cells. The components considered responsible for the adherence were recovered more from aminoglycoside treated cells than from control, benzylpenicillin or chloramphenicol treated cells. Using a contact angle measurement, the bacterial cell surface was found to be more hydrophobic after growing in the presence of some aminoglycosides, and hydrophilic which decreased adherence, after growing in the presence of benzylpenicillin and ofloxacin. Spectinomycin and kasugamycin, which are both aminoglycosides which do not cause misreading, failed to enhance adherence suggesting that misreading caused by aminoglycosides plays an important role in the enhancement of adherence. PMID:1816179

Miyake, Y; Kohada, A; Sugai, M; Suginaka, H

1991-12-01

 
 
 
 
141

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english MP [4-(3?,3?-dimethylallyloxy)-5-methyl-6-methoxyphthalide] was obtained from liquid culture of Pestalotiopsis photiniae isolated from the Chinese Podocarpaceae plant Podocarpus macrophyllus. MP significantly inhibited the proliferation of HeLa tumor cell lines. After treatment with MP, characterist [...] ic 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.

C., Chen; R.L., Yang.

2013-08-01

142

A 1H spin echo NMR study of HeLa tumour cell  

International Nuclear Information System (INIS)

Phosphorylcholine, phosphorylcreatine, lactate, glutathione, glycine and leucine were identified in living HeLa cells in the first study of this cell type by 1H spin echo NMR spectroscopy. The technique has the advantage that it is non-invasive, providing detailed structural information on individual species present in the cell matrix. It has been used in this case to study the rate of energy consumption following the activation of the glycolytic pathway with glucose. (Auth.)

143

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

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

144

Growth and apoptosis of HeLa cells induced by intense picosecond pulsed electric field  

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Full Text Available Objective To investigate the growth and apoptosis of HeLa cells induced by intense picosecond pulsed electric field(PEF in vitro.Methods HeLa cells cultured in vitro were divided into experimental group and control group(with or without intense picosecond PEF.With constant pulse width,frequency and voltage,the cells in experimental group were divided into 6 sub-groups according to the number of pulse(100,200,500,1000,1500,2000,the growth inhibition of HeLa cells by PEF and the dose-effect relationship were analyzed by MTT.Caspase 3 protein activity was detected in the cells in 500,1000 and 2000 sub-groups.Mitochondrial transmembrane potential was detected by rhodamine 123 staining with the cells in 2000 sub-groups.Results MTT assay demonstrated that intense picosecond PEF significantly inhibited the proliferation of HeLa cells in dose-dependent manner.The survival rates of cells declined along with the increase in pulse number,and were 96.23%±0.76%,94.11%±2.42%,90.31%±1.77%,64.59%±1.59%,32.95%±0.73%,23.85%±2.38% and 100%,respectively,in 100,200,500,1000,1500,2000 sub-groups and control group(P < 0.01.The Caspase 3 protein activity was significantly enhanced by intense picosecond PEF,and the absorbancy indexes(A were 0.174±0.012,0.232±0.017,0.365±0.016 and 0.122±0.011,respectively,in 500,1000,2000 sub-groups and control group(P < 0.05.The mitochondrial transmembrane potential of HeLa cells was significantly inhibited by intense picosecond PEF,and the fluorescence intensity in 2000 sub-group(76.66±13.38 was much lower than that in control group(155.81±2.33,P < 0.05.Conclusion Intense picosecond PEF may significantly inhibit the growth of HeLa cells,and induce cell apoptosis via mitochondrial pathway.

Yuan-yuan HUA

2011-07-01

145

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

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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

146

Diffusion kinetics of tritiated uridine into HeLa cells previously exposed to hyperthermia  

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Hyperthermic treatment of HeLa cells at 420C for 60 min depressed the specific activity of these cells when incubated with 3H-uridine both during and post heating compared to cells maintained at 370C. These changes were unlikely to arise from increased leakage from the cells and may partially be attributed to membrane damage influencing facilitated diffusion. Diffusion kinetic data for incorporation of the radiolabel into the T.C.A. soluble and T.C.A. insoluble fractions of HeLa cells indicated that a significant depression of Vsub(max) and a significant elevation of Ksub(m) for incorporation of 3H UdR into RNA may possibly result from an isotope dilution effect attributed to degrading pre-ribosomal RNA under the effect of hyperthermia. (author)

147

Proliferation inhibition and radiosensitization effects of 6-bromoisovanillin on HeLa cells  

International Nuclear Information System (INIS)

Objective: To study the effects of 6-bromoisovanillin (6-bromine-5-hydroxy-4-methoxy-benzaldehyde, BVAN08) on the proliferation and radiosensitivity of cultured HeLa cells, and to provide experimental data and advanced evidence for exploring anticancer and radiation-sensitizing new drugs. Methods: MTT assay was used to detect cell proliferation, the DNA damage was detected by single-cell gel electrophoresis. Flow cytometry was used to monitor cell cycle changes, colony-forming ability assay was used to analyze the radiosensitivity, and apoptosis was detected by AnnexinV/PI staining. Results: BVAN08 suppressed the proliferation of HeLa cells at the concentrations over 10-20 ?mol/L in a dose-dependent manner. The survival cure analysis indicated that the IC50 was 19.07 ?mol/L. DNA double-strand breaks were also detected in the cells after the treatment with BVAN08. BVAN08 initiated a G2/M phases arrest four hours after treatment. The accumulation of G2/M population increased with the duration of treatment. The expression of DNA repair protein DNA-PKcs was remarkably depressed by BVAN08. Combined treatment with 20 ?mol/L BVAN08 and 2 Gy 60Co ?-rays significantly decreased the cell survival and increased the induction of apoptosis. Conclusion: BVAN08 can inhibit proliferation and enhance the radiosensitivity of HeLa cells. The radiosensitization is related to the suppression of DNA-PKcs and G2/M phase arrest. (authors)

148

Adherence of Actinomyces pyogenes to HeLa cells mediated by hydrophobic surface proteins.  

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Determination of cell-surface hydrophobicity of Actinomyces pyogenes by hydrophobic interaction chromatography on phenyl-Sepharose revealed that all 42 cultures examined were strongly hydrophobic. The hydrophobic surface proteins were solubilized by mutanolysin treatment of the bacteria and isolated by hydrophobic interaction chromatography. In SDS-PAGE, they appeared with numerous protein bands and blocked the adhesion of whole bacterial cells to the gel matrix. The A. pyogenes cultures attached to HeLa cells in varying degrees. This attachment of A. pyogenes was greatly reduced in the presence of isolated hydrophobic proteins and in the presence of specific antibodies produced against hydrophobic surface proteins. The results of the present study demonstrate that hydrophobic surface proteins promote the capacity of A. pyogenes to adhere to HeLa cells. PMID:8219500

Ding, H; Lämmler, C; Seleim, R S

1993-08-01

149

DNA polymerases alpha, delta, and epsilon: three distinct enzymes from HeLa cells.  

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DNA polymerases alpha, delta, and epsilon have been purified and characterized from the same HeLa cell extract in order to determine their relationship by comparing them from the same cell type. The catalytic properties and the primary structures of the large subunits of the DNA polymerases as compared by partial peptide mapping with N-chlorosuccinimide are different. Likewise, the small subunit of DNA polymerase epsilon appears to be distinct from the large subunit of the same polymerase and...

Syva?oja, J.; Suomensaari, S.; Nishida, C.; Goldsmith, J. S.; Chui, G. S.; Jain, S.; Linn, S.

1990-01-01

150

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

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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

151

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

International Nuclear Information System (INIS)

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

152

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

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

Kezban Ada

2010-04-01

153

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

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

Nisa Tandogan

2011-04-01

154

Clonal heterogeneity in the progeny of HeLa cells which survive X-irradiation  

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Clones of HeLa cells that had survived 9 Gy X-rays were investigated for plating efficiency, radiosensitivity, the proportion of giant cells, and chromosome numbers. A significant decrease in plating efficiency was observed that persisted for > 20 population doublings. The reduced plating efficiency was associated with an increased proportion of giant cells in the colonies. There was pronounced clonal heterogeneity in the progeny of irradiated surviving cells with respect to plating efficiency, proportion of giant cells, and chromosome numbers. The radiosensitivity of progeny from irradiated cells did not change compared with controls. (author)

155

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

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Full Text Available The bacteriostatic activity of some of alkyl substituted (E-b-(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

156

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.

157

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 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.

158

Comparison of metaphase and interphase nucleolar activity in Hela-CCL2 cells and PHA-stimulated human lymphocytes.  

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Metaphase and interphase nucleolar activity in cultured Hela-CCL2 cells and PHA-stimulated human lymphocytes have been studied with silver nitrate staining. In metaphase, we examined the relationship between the actual number of active NORs (AgNORs) and total number of NOR-bearing acrocentrics. Interphase silver staining over the nucleus was analyzed cytodensitometrically and morphologically. From all investigations, Hela-CCL2 cells and lymphocytes were shown to have similar levels of nucleolar activity. Our results suggest that there is a form of regulation of nucleolar activity in malignant Hela-CCL2 cells as compared to PHA-stimulated human lymphocytes. PMID:6208996

Van der Elst, J; Deleener, A; Verschaeve, L; Kirsch-Volders, M; Susanne, C

1984-11-01

159

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)

160

Identification of the methylated ribosomal proteins in HeLa cells and the fluctuation of methylation during the cell cycle.  

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Methylated proteins from HeLa cell cytoplasmic ribosomes have been identified. At least seven proteins are methylated and four of them are mildly acidic. The nature of the methylated amino acid in each protein is presented. In synchronized HeLa cell culture, the extent of methylation for both subunits varies with the cell cycle. Methylation of the 40 S subunit occurs heavily in the late G1 phase whereas methylation of the 60 S subunit is most pronounced in the early S phase. PMID:629983

Chang, F N; Navickas, I J; Au, C; Budzilowicz, C

1978-03-29

 
 
 
 
161

Action of caffeine on x-irradiated HeLa cells. II. Synergistic lethality  

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Postirradiation treatment of HeLa S3 cells with 1 mM caffeine results in a marked diminution of the surviving fraction as scored by colony formation. The decrease is dose dependent; the effect of a 24-hour postirradiation treatment of a nonsynchronous population with caffeine is to change the terminal slope of the survival curve and its intercept. D0 is reduced from 130 to 60 rad; the extrapolation number is increased about twofold. The amount of postirradiation killing is maximal if cells are exposed to caffeine at a concentration of at least 1 mM for 8 hours; less than 10% of unirradiated cells are killed under these conditions. Dose-response curves were also obtained for synchronous cells at various phases of the cell cycle. Similar results were obtained at all cell ages, but the magnitude of the effect is age dependent. This age dependence was further explored in experiments in which mitotically collected cells were exposed to 300 or 500 rad doses at 2-hour intervals throughout the cell cycle. Treatment with caffeine for 24 hours after irradiation enchances the killing of cells late in the cycle more than cells in G1. The sensitivities of two other cell lines, CHO and EMT6, also were examined; both are substantially less sensitive to caffeine. The smaller cell-cycle dependence of CHO cells is qualitatively the same as that of HeLa cells

162

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

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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)

163

Trypanosoma cruzi trypomastigotes induce cytoskeleton modifications during HeLa cell invasion  

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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.

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

164

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

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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

165

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 5min 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 35nm. A zeta potential value of -27.6mV 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. PMID:24934968

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

2014-12-10

166

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

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

Kosti? Milena

2008-01-01

167

DNA polymerases. alpha. ,. delta. , and. var epsilon. : Three distinct enzymes from HeLa cells  

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DNA polymerases {alpha}, {delta}, {epsilon} have been purified and characterized from the same HeLa cell extract in order to determine their relationship by comparing them from the same cell type. The catalytic properties and the primary structures of the large subunits of the DNA polymerases as compared by partial peptide mapping with N-chlorosuccinimide are different. Likewise, the small subunit of DNA polymerase {epsilon} appears to be distinct from the large subunit of the same polymerase and from the smaller subunits of DNA polymerase {alpha}. HeLa DNA polymerase {delta} is processive only when HeLa proliferating cell nuclear antigen is present, whereas DNA polymerase {epsilon} is quite processive in its absence. Inhibitor and activator spectra of DNA polymerases {alpha}, {delta}, and {epsilon} also distinguish the three enzymes. These results and immunologic comparisons published elsewhere support the premise that HeLaDNA polymerases {alpha}, {delta}, and {epsilon} are distinct enzymes that have common properties with yeast DNA polymerases I, HI, and II, respectively.

Syvaeoja, J.; Suomensaari, S.; Goldsmith, J.S.; Chui, G.S.J.; Jain, S.; Linn, S. (Univ. of California, Berkeley (USA)); Nishida, C. (Univ. of California, San Francisco (USA))

1990-09-01

168

Automatic segmentation of HeLa cell images  

CERN Document Server

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

Urban, Jan

2011-01-01

169

Laser stimulation can activate autophagy in HeLa cells  

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For decades, lasers have been a daily tool in most biological research for fluorescent excitation by confocal or multiphoton microscopy. More than 20 years ago, cell photodamage caused by intense laser stimulation was noticed by generating reactive oxygen species, which was then thought as the main damage effect by photons. In this study, we show that laser stimulation can induce autophagy, an important cell lysosomal pathway responding to immune stimulation and starvation, without any biochemical treatment. Two different types of laser stimulations are found to be capable of activating autophagy: continuous scanning by continuous-wave visible lasers and a short-time flash of femtosecond laser irradiation. The autophagy generation is independent from wavelength, power, and scanning duration of the visible lasers. In contrast, the power of femtosecond laser is very critical to autophagy because the multiphoton excited Ca2+ dominates autophagy signaling. In general, we show here the different mechanisms of autophagy generation by such laser stimulation, which correspond to confocal microscopy and cell surgery, respectively. Those results can help further understanding of photodamage and autophagy signaling.

Wang, Yisen; Lan, Bei; He, Hao; Hu, Minglie; Cao, Youjia; Wang, Chingyue

2014-10-01

170

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

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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

171

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

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

172

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

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

A. Soeksmanto

2010-01-01

173

Immunotherapy: rAAV2 expressing interleukin-15 inhibits HeLa cell tumor growth in mice  

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Full Text Available Abstract Human interleukin-15 (hIL15 has anti-tumor activities, but it is not convenient for tumor treatment because of its short half-life. A gene therapy for mouse lung cancer using an adenovirus vector expressing IL15 has been reported. However, adenovirus vector-mediated gene therapy can provoke cellular toxicity and inflammatory reactions. The recombinant adenovirus-associated vector 2 (rAAV2 is safer due to minimal cellular toxicity and immune response. In order to demonstrate that gene therapy can be used safely and successfully for human cancer treatment, the rAAV2 expressing hIL15 gene (rAAV2-hIL15 is applied for human cervical cancer, HeLa cell, in this study. This study successfully demonstrates that rAAV2-hIL15 can express IL15 with bioactivities in vitro and in vivo. In conclusion, our studies show that human cervical cancers are inhibited on animal model with rAAV2-hIL15 treatment and provide a safer and important reference for human cancer gene therapy.

Hung Yu-Ting

2009-05-01

174

The cold-shock response in mammalian cells: investigating the HeLa cell cold-shock proteome  

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In recent years there have been a number of reports that suggest the sub-physiological (<37 °C) temperature in vitro culturing of mammalian cells can result in enhanced heterologous protein production. Despite these reports, the mechanisms by which mammalian cells respond to such conditions are largely unknown. We therefore set out to use a model in vitro culture HeLa cell system to begin investigating the cold-shock response in mammalian cell systems. Sub-physiological temperature cultiv...

Underhill, Miche?le F.; Smales, C. Mark

2007-01-01

175

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

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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

176

Vibrio fluvialis attachs to but does not enter Hela cell monolayers  

Scientific Electronic Library Online (English)

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

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

177

Vibrio fluvialis attachs to but does not enter Hela cell monolayers  

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

I. T. Carvalho

1994-06-01

178

Spontaneous and radiation induced cell death in HeLa S3 human carcinoma  

International Nuclear Information System (INIS)

Radiation biologists have classified radiation-induced cell death based on cell proliferative capacity to either mitotic or interphase death. Cytologists have revealed two morphologically and biochemically diverse forms of cell death, apoptosis and necrosis. While the knowledge of the former is already well exploited by radiologists, cell susceptibility to apoptosis and necrosis is still under investigation. We studied characteristics of spontaneous cell death, and dose dependence and time course of radiation-induced cell death of human uterine cervix epitheloid carcinoma HeLaS3 in culture. Cells were irradiated with 2-40 Gy of ?-rays. The effect on growth, viability, morphology and genomic DNA structure were followed 24-72 h after irradiation. Cell viability was evaluated by trypan-blue exclusion assay and cell morphology by in situ DNA staining with propidium iodide. Cell genomic DNA fragmentation pattern was determined by electrophoresis on 2% agarose gels. At all cell densities 25-35% cells were PI positive and their DNA was fragmented to a high molecular size (?20 kbp), but the internucleosomal ladder was not observed. A significant decrease in viability to 33% was observed 72 h post 40 Gy irradiation. It corresponded to 55% of PI positive cells. A smear of smaller DNA fragments (0.1-1 kbp), 24 h after 10-20 Gy irradiation was considered as proof that the dominant form of radiation-induced cell death was necrosis. It was concluded that the dominant form of radiation-induced cell death in HeLaS3 population was necrosis and the radiation dose which caused 50% of cell death after 72 h (termed ND50) was between 30-40 Gy. (author)

179

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

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

180

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

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

Natalia M. Vladimirova

2011-11-01

 
 
 
 
181

Intracellular imaging of HeLa cells by non-functionalized NaYF4 : Er3+, Yb3+ upconverting nanoparticles  

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We report on the efficient incorporation of non-functionalized NaYF(4) : Er(3+), Yb(3+) nanoparticles inside HeLa live cancer cells by direct endocytosis. The efficient two-photon excited near-infrared-to-visible upconversion fluorescence of these nanoparticles is then used to obtain high-contrast intracellular fluorescence images of single cells. These images reveal a redistribution of the nanoparticles inside the cell as the incubation time increases. Thus, non-functionalized NaYF(4) : Er(3+), Yb(3+) nanoparticles emerge as very promising fluorescence probes for real-time imaging of cellular dynamics.

Vetrone, Fiorenzo; Naccache, Rafik

2010-01-01

182

Study on the characteristics of cell-cycle perturbation in hela cell exposed to continuous ? irradiation of 32P  

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In an attempt to understand radiobiological basis for targeted radiotherapy in oncology, the cell cycle perturbations have studied in Hela cell lines after exposed to different doses and dose-rate of 32P radiation. Asynchronous Hela cells, cultured in vitro, were exposed to ? radiation from radioactive filter papers (absorbed 32P) which were put close under culture plate of growing monolayer of Hela cells. The characteristic radiation response to different dose, dose-rate and radiation time was evaluated through cell cycle perturbation studied by flow cytometry. Cell cycle status showed G2 phase blockage in a way of dose dependence, a plateau of G2 block can be recognized at about 24h. Interestingly, the G2 phase declined even though the accumulated doses increased as the time of radiation prolonged. This result suggested that the cell cycle progress could not be inhibited completely when exposed to continuous radiation, rather it seems to be controlled somehow by the nature of cell cycle itself for a certain cell line. G2 blockage, one of the major changes caused by ? radiation, is dose-dependent, but the time reaching the plateau of G2 phase blockage is most likely related with the intrinsic nature of cell cycle

183

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)

184

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

185

Mitomycin-induced chromatid breaks in HeLa cells: a consequence of incomplete DNA replication.  

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The formation of chromosome aberrations induced by alkylating agents such as mitomycin C has been shown to require the passage of the treated cell through S phase. However, the exact mechanisms by which mitomycin C-induced DNA lesions are translated into chromosome aberrations during S phase are not known. The purpose of these studies was to better understand the molecular basis of chromosome aberration formation after mitomycin C treatment. The morphology of metaphases of cells treated in G1 phase with mitomycin C resembled that of prematurely condensed chromosomes of S-phase cells. Consequently we postulated that chromosome aberrations resulted from cells reaching mitosis without completing DNA replication. This was tested by treating HeLa cells in G1 phase with mitomycin C and then analyzing these cells at mitosis for residual DNA damage and DNA content. Utilizing the DNA alkaline elution assay for DNA damage, we showed that HeLa cells progress through S phase into mitosis with intact DNA-DNA interstrand cross-links. These cross-links, originally induced into parental DNA, were associated equally with parental and newly replicated DNA at the time the cells reached mitosis. This suggests that recombinational events had taken place during the DNA replication process. Cells that were treated in G1 phase and allowed to proceed to mitosis in the presence of bromodeoxyuridine to density label newly replicated DNA were analyzed with cesium chloride density sedimentation. Unreplicated DNA was present in the mitotic cells of the treated populations but not in the untreated control cells. Further, flow cytometric measurements, made under hypotonic conditions in order to reduce chromatin condensation effects, demonstrated that the mitotic cells from the mitomycin C-treated populations contained 10-20% less DNA than untreated mitotic controls. These results indicate that chromosome breaks induced by mitomycin C are the result of cells reaching mitosis without having fully completed DNA replication. PMID:3089585

Sognier, M A; Hittelman, W N

1986-08-01

186

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

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

187

Rose Bengal Acetate PhotoDynamic Therapy (RBAc-PDT) Induces Exposure and Release of Damage-Associated Molecular Patterns (DAMPs) in Human HeLa Cells  

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The new concept of Immunogenic Cell Death (ICD), associated with Damage Associated Molecular Patterns (DAMPs) exposure and/or release, is recently becoming very appealing in cancer treatment. In this context, PhotoDynamic Therapy (PDT) can give rise to ICD and to immune response upon dead cells removal. The list of PhotoSensitizers (PSs) able to induce ICD is still short and includes Photofrin, Hypericin, Foscan and 5-ALA. The goal of the present work was to investigate if Rose Bengal Acetate (RBAc), a powerful PS able to trigger apoptosis and autophagy, enables photosensitized HeLa cells to expose and/or release pivotal DAMPs, i.e. ATP, HSP70, HSP90, HMGB1, and calreticulin (CRT), that characterize ICD. We found that apoptotic HeLa cells after RBAc-PDT exposed and released, early after the treatment, high amount of ATP, HSP70, HSP90 and CRT; the latter was distributed on the cell surface as uneven patches and co-exposed with ERp57. Conversely, autophagic HeLa cells after RBAc-PDT exposed and released HSP70, HSP90 but not CRT and ATP. Exposure and release of HSP70 and HSP90 were always higher on apoptotic than on autophagic cells. HMGB1 was released concomitantly to secondary necrosis (24 h after RBAc-PDT). Phagocytosis assay suggests that CRT is involved in removal of RBAc-PDT generated apoptotic HeLa cells. Altogether, our data suggest that RBAc has all the prerequisites (i.e. exposure and/or release of ATP, CRT, HSP70 and HSP90), that must be verified in future vaccination experiments, to be considered a good PS candidate to ignite ICD. We also showed tha CRT is involved in the clearance of RBAc photokilled HeLa cells. Interestingly, RBAc-PDT is the first cancer PDT protocol able to induce the translocation of HSP90 and plasma membrane co-exposure of CRT with ERp57. PMID:25140900

Panzarini, Elisa; Inguscio, Valentina; Fimia, Gian Maria; Dini, Luciana

2014-01-01

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

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

189

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

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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?.

190

Clonal variability of radiation-induced cisplatin resistant HeLa cells.  

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Low-dose fractionated gamma-irradiation (3 cycles of 5x2 Gy) induces moderate cisplatin resistance in HeLa cells which is associated with alterations of the caspase-dependent apoptotic pathway (2). There is a considerable heterogeneity in cell survival among isolated resistant clones (Rf values between 1.4 to 2.4) and in the propensity of the cells to enter apoptosis. These variations are associated with altered activation of the apoptotic pathway by members of the TNF family. The membrane receptor CD95 (Apo-1/Fas), which is upregulated immediately after cisplatin exposure in parental HeLa cells, is expressed at various levels in the resistant clones. There are also changes in the formation of the inhibitor protein I kappa B, which regulates the antiapoptotic transcription factor NF kappa B. Since the response to radiation is unchanged, the results collectively suggest that changes in the activation of the caspase-dependent signalling cascade are involved in the death pathway initiated by cisplatin but not by radiation damage. PMID:9713497

Eichholtz-Wirth, H; Marx, K

1998-01-01

191

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)

192

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

193

Negative pion depth-dose profile examined by means of HeLa cell survival curves  

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HeLa cells were irradiated at liquid nitrogen temperature with negative pions from Nimrod at the Rutherford Laboratory and then assayed for survival of colony-forming ability. Complete dose-response curves were obtained from repeated determinations at fourteen different positions along the depth dose profile and survival curves fitted to the data by computer programme. A depth damage profile was thus established in terms of the final slopes of these curves. This confirmed the expected RBE value of the 1.9 at the ionisation peak. Although a value of unity was found in the main plateau region, the 'entrance' positions showed significantly higher values. (author)

194

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

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

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

1984-02-01

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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

196

Suppressive effect on HeLa cells proliferation by phenothiazine derivatives alone and combining with ionizing radiation  

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Objective: To examine the antiproliferative effects of phenothiazine derivatives (PTZDs) alone on HeLa cells and in combination with ionizing radiation. Methods: MTT and colony-forming method were used to evaluate the proliferation activity and cellular radiosensitivity of HeLa cells. Results: We compared the antiproliferative effects of six phenothiazine derivatives, and found that the derivatives ?-chloro-N-dimethylamine phenothiazine (PTZD2), ?-triflumethyl-N-?(dimethylamine ethyl) phenothiazine (PTZD3) and ?-chloro-N-(dimethylamine ethyl) phenothiazine (PTZD5) showed a significant antiproliferative effect at concentration of 10 ?mol/L. HeLa cells proliferation was completely suppressed when treated with PTZDs of 40-50 ?mol/L. PTZD2/PTZD3 and cobalt-60 gamma-irradiation showed synergistic suppressive effect on proliferation of HeLa cells. The enhancement ratios of 10 ?mol/L PTZD3 combination with 2 Gy and 4 Gy irradiations were 3.5 and 1.8, respectively. The maximum synergistic suppressive effect was observed when cells administered with PTZD3 at 18 h before being irradiated. Conclusion: Phenothiazine derivatives show antiproliferations on HeLa cells, and differ in degrees. The synergistic anticancer effect could be obtained by combining phenothiazine derivatives with radiotherapy. (authors)

197

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

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

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

2010-01-25

198

Cytotoxic Activity of Three South Sulawesi Medicinal Plant Extracts Used in the Treatment of HeLa Cell Line: Jati Putih (Gmelina arborea Roxb., Jati Belanda (Guazuma ulmifolia Lamk. and Lakkalakka (Curculigo orchioides Gaerth  

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Full Text Available Gmelina arborea Roxb, Guazuma ulmifolia Lamk and Curculigo orchioides Gaerth, the three plants frequently used in South Sulawesi for the treatment of cancerous diseases, have been selected to examine their action in cervical epithelial carcinoma. These extracts were assessed using HeLa cell cancer (Human cervix cancer and doxorubicin was used as the positive control. Data are presented as the dose that inhibited 50% control growth (IC50. Cytotoxic activity was measured using MTT colorimetric assay. Dose-dependent studies revealed IC50 of 113.61±0.12 ?g/mL, 174.90±1.22 ?g/mL and 126.05±2.43 ?g/mL for eGA, eGU and eCO on HeLa cell cancer, respectively and correlated with treatment of cancer.

LUKMAN M

2014-09-01

199

Degradation of structurally characterized proteins injected into HeLa cells. Basic measurements  

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Thirty-five proteins of known x-ray structure were labeled by chloramine-T radioiodination or by reaction with 125I-Bolton-Hunter reagent and introduced into HeLa cells using red cell-mediated microinjection. Degradation rates of the injected proteins were then determined over the next 50 h by measuring the release of soluble isotope to the culture medium. Control experiments demonstrated that the measured rates were not compromised by proteolysis within RBCs, the presence of unfused RBCs, or degradation of protein released from RBCs to the medium. Degradation of some injected proteins was faster during the first 12 h after fusion than at later times, apparently a response of HeLa cells to trypsinization. However, all proteins exhibited first-order degradation rates between 24 and 48 h post injection. Except for seven proteins, stabilities measured during this interval were unaffected by the labeling procedure. Reductive methylation was used to choose among the seven discordant values, and half-lives for the 35 proteins ranged from 16 h for lysozyme to 214 h for yeast alcohol dehydrogenase. Since half-lives for six of the injected proteins closely match values obtained by in vivo measurements, we consider our estimates of the metabolic stabilities of the injected proteins to be generally accurate. Therefore, the half-lives obtained by microinjection should prove useful in the search for relationships between protein structure and intracellular stabilityand intracellular stability

200

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

 
 
 
 
201

Investigation of siRNA Nanoparticle Formation Using Mono-Cationic Detergents and Its Use in Gene Silencing in Human HeLa Cells  

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The focus of recent research has been on the development of siRNA vectors to achieve an innovative gene therapy. Most of the conventional vectors are siRNA nanoparticles complexed with cationic polymers and liposomes, making it difficult to release siRNA. In this study, we report on the use of MCD, a quaternary ammonium salt detergent containing a long aliphatic chain (L-chain) as an siRNA complexation agent using human HeLa cells (a model cancer cell). We prepared siRNA nanoparticles using v...

Hideyoshi Harashima; Yuma Yamada; Ryosuke Suzuki

2013-01-01

202

Involvement of the Nrf2 Pathway in the Regulation of Pterostilbene-Induced Apoptosis in HeLa Cells via ER Stress.  

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Among the various cancer cell lines, HeLa cells were found to be sensitive to pterostilbene (Pte), a compound that is enriched in small fruits such as grapes and berries. However, the mechanism involved in the cytotoxicity of Pte has not been fully characterized. Using biochemical and free radical biological experiments in vitro, we identified the pro-apoptotic profiles of Pte and evaluated the level of redox stress-triggered ER stress during HeLa cell apoptosis. The data showed a strong dose-response relationship between Pte exposure and the characteristics of HeLa apoptosis in terms of changes in apoptotic morphology, DNA fragmentation, and activated caspases in the intrinsic apoptotic pathway. During drug exposure, alterations in the intracellular redox homeostasis that favor oxidation were necessary to cause ER stress-related apoptosis, as demonstrated by enzymatic and non-enzymatic redox modulators. A statistically significant and dose-dependent increase (P anti-oxidant application with the Pte treatment. Our research demonstrated that Pte trigged ER stress by redox homeostasis imbalance, which was negatively regulated by a following activation of Nrf2. PMID:25341683

Zhang, Bo; Wang, Xiao-Qin; Chen, Han-Yin; Liu, Bin-Hua

2014-11-20

203

Differential effect of procaine on irradiated mammalian cells in culture. [. gamma. rays; HeLa cells; V-79 Chinese hamster cells  

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HeLa and V-79 Chinese hamster cells temporarily stored in ampoules were treated with the local anesthetic procaine. Postirradiation treatment increased lethality in HeLa cells depending on drug concentration, duration of treatment, and cell density, as measured by colony-forming ability upon plating. If present during irradiation only, procaine protected from irradiation. In V-79 cells, procaine potentiated radiation lethality only in freshly trypsinized cells. Procaine effect was thus cell type specific and most likely involved the cell membrane.

Djordjevic, B.

1979-05-01

204

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

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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)

205

Changes in the protein-synthesizing system of HeLa cells in culture in the presence of trace elements  

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This paper studies the state of the protein-synthesizing system of HeLa cells in culture in the presence of certain trace elements. The cytopathic action of zinc, nickel, cobalt, cadmium, and fluorine was studied in the presence of maximal allowable concentrations adopted for liquid media. Thirty minutes before the end of incubation with the elements to be studied, (H 3)-uridine or (H 3)-leucine was added to the cultures of HeLa cells. The autoradiographic data showed that variation in the integral parameters of cell function as the level of synthesis of total fast-labeled RNA and total protein in fact do take place during incubation of the HeLa cell culture with trace elements

206

Prevotella bivia can invade human cervix epithelial (HeLa) cells.  

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Prevotella bivia has been associated with female upper genital tract infections and an increased risk of preterm delivery. In this study, the adherence and invasion capacity of P. bivia was investigated using a cervix epithelial cell line. P. bivia was furthermore analysed for its ability to evoke a proinflammatory cytokine response in epithelial cells. The invasion capacity, defined as the number of bacteria recovered from lysed HeLa cells infected with P. bivia, varied considerably among five strains, all of which were isolates from women with bacterial vaginosis. One P. bivia strain (P47) gave rise to an approximately 120-fold higher number of intracellular bacteria (7 x 10(3) bacteria per 1 x 10(5) cells) compared with the least invasive strain. Three strains expressed an intermediate or low invasiveness, showing an approximately 3- to 40-fold higher number of intracellular bacteria per 1 x 10(5) cells compared with the least invasive strain. The intracellular localization of P47 in phagosome-like vesicles was confirmed by transmission electron microscopy. All P. bivia strains adhered to HeLa cells to the same extent (range 14-22 bacteria per cell) as analysed by interference microscopy. No correlation was found between adhesion and invasion capacity of the strains. Furthermore, no fimbriae-like structures were observed on P47 detected by scanning electron microscopy or negative staining. Analysis of TNF-alpha, IL-1alpha, IL-6, IL-8, and IL-18 in P. bivia-stimulated HeLa cells showed low levels of only IL-6 and IL-8 for the most invasive P. bivia strain P47. Thus, the induction of IL-6 or IL-8 secretion appeared to be associated with invasion capacity. This work provides evidence that some P. bivia isolates can invade human cervix epithelial. Thus, a strong capacity for invasion and a weak proinflammatory cytokine-inducing capacity in P. bivia are suggested to be virulence factors in establishing a low-grade upper genital tract infection. PMID:17367470

Strömbeck, Louise; Sandros, Jens; Holst, Elisabeth; Madianos, Phoebus; Nannmark, Ulf; Papapanou, Panos; Mattsby-Baltzer, Inger

2007-03-01

207

Mechanistic aspects of fluorescent gold nanocluster internalization by live HeLa cells  

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

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

2013-01-01

208

Gefitinib Inhibits the Growth of Toxoplasma gondii in HeLa Cells.  

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Toxoplasma gondii is the causative agent of toxoplasmosis with symptoms of congenital neurological and ocular diseases and acquired lymphadenitis, retinochoroiditis, and meningoencephalitis. Small molecules which block the activity of protein kinases were tested in in vitro culture of T. gondii to find new therapeutic drugs of safer and more effective than the combined administration of pyrimethamine and sulfadoxine that sometimes provoke lethal Stevens-Johnson syndrome. Among them, Gefitinib and Crizotinib inhibited intracellular growth of T. gondii in HeLa cells by counting the number of T. gondii per parasitophorous vacuolar membrane whereas Sunitinib did not. Gefitinib inhibited the growth of T. gondii in a dose-dependent manner over 5 µM up to the tolerable concentration of HeLa cells and halted the division of the parasite immediately from the time point of treatment. Gefitinib inhibition suggests that tyrosine kinases of EGFR family or other homologous kinases of the parasite itself may be the target to cause the block of T. gondii growth. PMID:25246725

Yang, Zhaoshou; Ahn, Hye-Jin; Nam, Ho-Woo

2014-08-01

209

Gefitinib Inhibits the Growth of Toxoplasma gondii in HeLa Cells  

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Toxoplasma gondii is the causative agent of toxoplasmosis with symptoms of congenital neurological and ocular diseases and acquired lymphadenitis, retinochoroiditis, and meningoencephalitis. Small molecules which block the activity of protein kinases were tested in in vitro culture of T. gondii to find new therapeutic drugs of safer and more effective than the combined administration of pyrimethamine and sulfadoxine that sometimes provoke lethal Stevens-Johnson syndrome. Among them, Gefitinib and Crizotinib inhibited intracellular growth of T. gondii in HeLa cells by counting the number of T. gondii per parasitophorous vacuolar membrane whereas Sunitinib did not. Gefitinib inhibited the growth of T. gondii in a dose-dependent manner over 5 µM up to the tolerable concentration of HeLa cells and halted the division of the parasite immediately from the time point of treatment. Gefitinib inhibition suggests that tyrosine kinases of EGFR family or other homologous kinases of the parasite itself may be the target to cause the block of T. gondii growth. PMID:25246725

Yang, Zhaoshou; Ahn, Hye-Jin

2014-01-01

210

PVA engineered microcapsules for targeted delivery of camptothecin to HeLa cells  

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Capsular microvectors are an important tool in the recent research field of nanomedicine to address a drug cargo for the therapeutic treatment of several pathologies. In this study we describe how the product of the conjugation of the polysaccharide chitosan with folate can be used as a coating of poly (vinyl alcohol), PVA, based microcapsules for an efficient targeting of HeLa cells. The influence of the coating on the bioadhesive properties of the vector and on its cargo capacity was also considered using camptothecin as an anticancer drug model. The coating strategy was finalized to exploit the good chemical versatility of PVA, used to form the shell of the vector. This study is a follow up of an investigation activity aiming to show the potentialities of PVA-shelled microcapsules or microbubbles as injectable microdevices supporting a theranostic approach for different types of tumour. Highlights: {yields}Coating of PVA-shelled microcapsules with chitosan-folate. {yields} Selective bioadhesion of microcapsules to HeLa Cells. {yields} Effective loading and release of camptothecin. {yields} In vitro anti-proliferative action of camptothecin loaded microcapsules.

Galbiati, Alice; Rocca, Blasco Morozzo della; Tabolacci, Claudio; Beninati, Simone; Desideri, Alessandro [Dipartimento di Biologia, Universita di Roma Tor Vergata, Via della Ricerca Scientifica, 00133 Rome (Italy); Paradossi, Gaio, E-mail: paradossi@stc.uniroma2.it [Dipartimento di Scienze e Tecnologie Chimiche, Universita di Roma Tor Vergata, Via della Ricerca Scientifica, 00133 Rome (Italy)

2011-12-01

211

Biostimulation of HeLa cells by low-intensity visible light. Pt. 5  

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Autoradiographic experiments, performed with monolayer HeLa cells, show that the irradiation with He-Ne laser (d=100 J/m2) causes an increase in the number of S-phase cells connected with enhanced G1-S transition of a part of the population, as well as an increase in the grain count on the labelled nuclei connected with an enhancement of DNA synthesis in S-phase cells. The irradiation influences the proliferation rate of various subpopulations not in equal degree, as shown analysing the clone size distribution after the irradiation (D 10, 102, 103 J/m2). The stimulative effect of irradiation is most noticeable on the proliferation activity of the slowly growing subpopulations

212

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

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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

213

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

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

E. Garcia-Alcantara

2013-07-01

214

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

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

215

LyGDI expression in HeLa cells increased its sensitivity to radiation-induced apoptosis  

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Objective: In order to confirm whether LyGDI has apoptotic signal transduction function and can increase the apoptotic rate of radiation-induced cell death, the lyGDI and mutant D19lyGDI gene, which constructed with the pCDNA3. 1 His A, were transfected into no-endogenous lyGDI HeLa cells. Methods Transient expressions of lyGDI and D19lyGDI in HeLa cells were analyzed by Western blot using anti-mono antibody of LyGDI and Xpress tag. Cell apoptosis was assayed with Annexin V-FITC apoptosis kit. To select stable clone, the transferred HeLa cells had been maintained in G418 medium for 3 weeks, then a cell line, which stably expressed LyGDI and mutant D19lyGDI, was selected. The selected cell line was irradiated with 12 Gy 60Co y-rays. Caspase-3 activity of the cells was determined by Western blot and cell viability by clone-forming assay after 48 hours post-irradiation culture. Results: Western blot and Annexin V-FITC apoptotic analysis revealed that lyGDI and D19lyGDI transient expressions in HeLa cells induced apoptosis; Caspase-3 activity measurement and clone-forming assay showed that lyGDI increased sensitivity to radiation-induced cell apoptosis. Conclusions: lyGDI performs function in apoptosis signal transduction, its expression in HeLa cells can increase the sensitivity to radiation-induced cell apoptosis. (authors)

216

Downregulation of FoxM1 inhibits proliferation, invasion and angiogenesis of HeLa cells in vitro and in vivo.  

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FoxM1 is a specific transcription factor that has an important function in aggressive human carcinomas, including cervical cancer. However, the specific function and internal molecular mechanism in cervical cancer remain unclear. In this study, RNAi-mediated FoxM1 knockdown inhibited cell growth. This process also decreased the migration and invasion activities of HeLa cells in vitro. Downregulation of FoxM1 inhibited tumor growth and angiogenesis in vivo. In addition, the expressions of uPA, matrix metalloproteinase (MMP)-2, MMP-9 and VEGF were significantly decreased in vitro and in vivo. These results suggested that the inactivation of FoxM1 could be a novel therapeutic target for cervical cancer treatment. PMID:25201015

Chen, Hong; Zou, Yang; Yang, Hong; Wang, Jingjing; Pan, Hong

2014-12-01

217

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)

218

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 L4. 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

219

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.

220

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

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

ANA NICIFOROVIC

2007-10-01

 
 
 
 
221

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

222

Prolonged cell cycle response of HeLa cells to low-level alkylation exposure  

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Alkylation chemotherapy has been a long-standing treatment protocol for human neoplasia. N-methyl-N’–nitro-N-nitrosoguanidine (MNNG) is a direct-acting monofunctional alkylator. Temozolomide is a clinical chemotherapeutic equivalent requiring metabolic breakdown to the alkylating agent. Both chemicals have similar mechanistic efficacy against DNA mismatch repair proficient tumor cells that lack expression of methylguanine methyltransferase (MGMT). Clinically relevant concentrations of both agents affect replicating cells only after the first cell cycle. This phenomenon has been attributed to replication fork arrest at unrepaired O6methyldeoxyguanine lesions mispaired with thymine during the first replication cycle. Here we demonstrate, by several different approaches, that MNNG-treated tumor cells do not arrest within the second cell cycle. Instead the population slowly traverses through mitosis without cytokinesis into a third cell cycle. The peak of both ss- and ds-DNA breaks occur at the height of the long mitotic phase. The majority of the population emerges from mitosis as multinucleated cells that subsequently undergo cell death. However, a very small proportion of cells, less than 1:45,000, survive to form new colonies. Taken together, these results indicate that multinucleation within the third cell cycle, rather than replication fork arrest within the second cell cycle, is the primary trigger for cell death. Importantly, multinucleation and cell death is consistently avoided by a small percentage of the population that continues to divide. This information should prove clinically relevant for the future design of enhanced cancer chemotherapeutics. PMID:19638578

Schroering, Allen G.; Kothandapani, Anbarasi; Patrick, Steve M.; Kaliyaperumal, Saravanan; Sharma, Vishal P.; Williams, Kandace J.

2009-01-01

223

Prolonged cell cycle response of HeLa cells to low-level alkylation exposure.  

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Alkylation chemotherapy has been a long-standing treatment protocol for human neoplasia. N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) is a direct-acting monofunctional alkylator. Temozolomide is a clinical chemotherapeutic equivalent requiring metabolic breakdown to the alkylating agent. Both chemicals have similar mechanistic efficacy against DNA mismatch repair-proficient tumor cells that lack expression of methylguanine methyltransferase. Clinically relevant concentrations of both agents affect replicating cells only after the first cell cycle. This phenomenon has been attributed to replication fork arrest at unrepaired O(6)-methyldeoxyguanine lesions mispaired with thymine during the first replication cycle. Here, we show, by several different approaches, that MNNG-treated tumor cells do not arrest within the second cell cycle. Instead, the population slowly traverses through mitosis without cytokinesis into a third cell cycle. The peak of both ssDNA and dsDNA breaks occurs at the height of the long mitotic phase. The majority of the population emerges from mitosis as multinucleated cells that subsequently undergo cell death. However, a very small proportion of cells, <1:45,000, survive to form new colonies. Taken together, these results indicate that multinucleation within the third cell cycle, rather than replication fork arrest within the second cell cycle, is the primary trigger for cell death. Importantly, multinucleation and cell death are consistently avoided by a small percentage of the population that continues to divide. This information should prove clinically relevant for the future design of enhanced cancer chemotherapeutics. PMID:19638578

Schroering, Allen G; Kothandapani, Anbarasi; Patrick, Steve M; Kaliyaperumal, Saravanan; Sharma, Vishal P; Williams, Kandace J

2009-08-01

224

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

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

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

2013-05-01

225

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

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

226

Cell cycle dependency of FDG and 67Ga citrate uptake in HeLa S 3 cells  

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Positron emission tomography using fluorine-18 fluoro-deoxyglucose (FDG PET) is useful for the detection of malignant tumor recurrences and for the evaluation of a therapeutic response to these; including ones found in the lung, colon, head and neck regions. The experimental study demonstrated FDG uptake was higher in faster-growing rather than in slower-growing tumors. These findings show FDG accumulation exhibits cell cycle dependency. However, the precise mechanism remains to be elucidated. In this study, the relationship between FDG uptake and the cell cycle phase in HeLa S 3 cells, as well as how they compare to the conventional tracer 67Ga citrate (Ga) with single photon emission tomography (SPECT) was assessed. Synchronization of HeLa S 3 cells was accomplished via a double thymidine block. The uptake of FDG and Ga was determined after cell cycle synchronization. The glucose transporter was independently evaluated for the level of its Glut 1 glucose transporter membrane protein. FDG uptake in HeLa S 3 cells was significantly higher in the early S phase and G2/M phase compared to the G1 phase. In addition, Ga uptake was higher in the G2/M phase. Immunochemical assays for the Glut 1 transporter showed an increase in membrane expression within S phase cells. It has been concluded that cell cycle dependency is reflected in the uptake of FDG and Ga, seen during PET or SPECT imaging of tumor tissue. These results reveal tumor proliferative activity, and can assist in evaluating a therapeutic response. Furthermore, the increased FDG uptake, in part due to increased membrane expression of the Glut 1 glucose transporter, contributes to this phenomenon. (author)

227

On enhancing drugs effect on radiosensitivity of HeLa cells by inhibiting P13K/Akt signal transduction  

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Objective: To explore the mechanism of PI3K/Akt in radiosensitization of docetaxel and cisplatin by inhibiting PI3K/Akt pathway in HeLa cells. Methods: To detect the 50% inhibition concentration (IC50) of cisplatin and docetaxel in Hela cells by mono-nuclear cell direct cytotoxicity assay (MTT) in vitro. Using the IC20 of cisplatin and docetaxel in Hela cell or in association with LY294002 for 24 h, then, the cells were irradiated by X-ray with 2,3,4,6,8 Gy. The cell survival fraction was computed by clone formation. Cell survival curve was fitted by multitarget one-hit model, and Dq, D0, SF2, sensitizing enhancing ratio(SER) was calculated. The expression of pAkt and total Akt by western blot were detected. Apoptosis was detected by flow cytometry. Results: 1. Docetaxel and cisplatin improved the phosphorylation of Akt by irradiation obviously. 2. The SER of docetaxel + LY294002 + irradiation group (1.92) was higher than that of docetaxel + irradiation group(1.41). The SER of cisplatin + LY294002 + irradiation group(1.71) was higher than the cisplatin + irradiation group (1.37). 3. Apoptosis rate of docetaxel + LY294002 + irradiation and cisplatin + LY294002 + irradiation groups(12.5%, 10.2%) were higher than those of docetaxel + irradiation and cisplatin + irradiation groups(6.1%, 5.1%). Conclusions: PI3K/Akt signal transduction activation may be as an important reason of radiosensitization reduction of docetaxel and cisplatin in the HeLa cells. Our results show that inhibiting PI3K/Akt can improve the radiosensitization of docetaxel and cisplatin in the HeLa cells. (authors)

228

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

229

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

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

230

Interaction of Enteropathogenic or Enterohemorrhagic Escherichia coli with HeLa Cells Results in Translocation of Cortactin to the Bacterial Adherence Site  

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Infection of cultured HeLa epithelial cells with enteropathogenic Escherichia coli (EPEC) or enterohemorrhagic E. coli (EHEC) O157:H7 results in accumulation of cortactin under the adherent bacteria. Tyrosine phosphorylation of cortactin is not induced following HeLa cell infection with EHEC or EPEC, contrary to what has been reported to occur with Shigella flexneri.

Cantarelli, Vlademir V.; Takahashi, Akira; Akeda, Yukihiro; Nagayama, Kenichi; Honda, Takeshi

2000-01-01

231

High temperature enhances cytotoxicity of mercury (HgCl2) on Hela S3 cells  

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The combined effect of mercury (HgCl2) and high temperature on the growth and synthesis of nucleic acid and protein, and on the cell cycle of HeLa S3 cells was investigated. The subsequent growth of the cells was dose-dependently inhibited by mercury at 37.2° and 41.2°C. The inhibitory effect of mercury on subsequent growth was enhanced at the higher temperature. IC50 values for DNA and RNA synthesis but not protein synthesis, at 41.2°C, were significantly lower than those at 37.2°C ( P<0.05, P<0.01, respectively). Flow cytometric analysis using synchronous cells indicated the possibility of blocking of cell cycle progression in the early part of S phase by the combined treatment. These results suggest that the cytotoxicity of mercury to cell growth was enhanced at the higher temperature and that this enhancement is related to the increased inhibitory effect of mercury on DNA and RNA synthesis and on the cell cycle at high temperatures.

Kishimoto, Takuji; Fukuzawa, Yoichiro; Tada, Manabu

1990-09-01

232

Mesoporous silica nanoparticles enhance MTT formazan exocytosis in HeLa cells and astrocytes.  

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We report on the observation that mesoporous silica nanoparticles (MSNs), after being endocytosed, interfere with the MTT test in HeLa cells and astrocytes by accelerating the exocytosis of formazan crystals. The stimulation of MTT formazan exocytosis is probably related to perturbation of intracellular vesicle trafficking by MSN uptake as revealed by experiments in presence of chloroquine and genistein. Similar effect has been previously observed with a number of chemicals, especially with neurotoxic beta amyloid peptides, but not with nanoparticles. We showed also that MTT reduction test gives an overestimation of the cytotoxicity of mesoporous silica nanoparticles compared to other tests such as LDH activity, WST-1 test and flow cytometry. These findings show that MTT assay should not be used for the study of MSN toxicity, and that perturbation of intracellular trafficking has to be taken into account in evaluating biocompatibility of MSNs. PMID:19254755

Fisichella, Matthieu; Dabboue, Hinda; Bhattacharyya, Sanjib; Saboungi, Marie-Louise; Salvetat, Jean-Paul; Hevor, Tobias; Guerin, Martine

2009-06-01

233

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-?-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; Wu, Jiazhen

2012-01-01

234

Physicochemical properties of the cytoplasmic glucocorticoid receptor complex in HeLa S3 cells.  

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The physiochemical properties, size, shape, and surface charge, have been determined for the cytoplasmic glucocorticoid receptor complex (GR) in randomly growing HeLa S3 cells. Sucrose density gradient and gel exclusion chromatographic analysis have shown GR to undergo a marked reduction in size and shape when analyzed under increasing KCl concentration conditions. Analysis of GR prepared in a hypotonic buffer solution revealed a large, 7-8 S species with Ve/Vo ratio of 1.15, and a Stokes radius of congruent to 95 A. Increasing the KCl concentration in the analysis buffer to 0.15 resulted in reduction in GR size and shape to a 4.5 S species with the following properties: Ve/Vo = 1.38. Stokes radius = 69 A. and calculated Svedberg molecular weight = 132,000. A limit to the observed decrease in GR size and shape was obtained under hypertonic, 0.4 M KCl, conditions; a 3.75 S form was observed to elute from the gel exclusion column with a Ve/Vo = 1.40. Stokes radius = 65 A. and a calculated Svedburg molecular weight = 102,000. Exposure to higher KCl conditions, 0.6 M and 0.8 M, resulted in no further decrease in GR size or shape. Ion exchange chromatographic analysis of cytoplasmic GR revealed heterogeneous populations of GR with apparent differences in surface charge. GR binding to DEAE cellulose revealed a predominant form which eluted at 0.15 M KCl (Form I). Under both hypotonic and hypertonic conditions small populations of GR forms were observed to elute from DEAE at or approximately 0.1 M KCl (Form III), and or approximately 0.3 M KCl (Form II), respectively. Binding of GR to hydroxylapatite (HAP) confirmed the heterogenous nature of GR. Cytoplasmic GR partitioned into at least 3 forms om HAP. The predominant GR form eluted from HAP at 0.1 M K2HPO4 (Form I); secondary, and tertiary forms eluted at 0.125 M (Form II), and 0.15 M K2POH4 (Form III), respectively. Glucocorticoid receptors were observed to elute from columns of phosphocellulose, and DNA cellulose in the void volume. Two receptor forms were observed with isoelectric focusing of HeLa S3 cytoplasm in Sephadex IEF G-75 gels. The primary, and secondary species had pI values of 7.5 and 6.3, respectively. These results demonstrate that randomly growing HeLa S3 cells contain heterogeneous population of GR in the cytoplasmic compartment. Randomly growing HeLa S3 cell cytoplasm appears to contain two "unactivated" receptor species (Forms I and II) that differ in their overall surface charge properties; a third activated species (Form III) appears to arise from heat induced "activation" during cell homogenization, and is more basic in nature. PMID:7087470

Currie, R A; Cidlowski, J A

1982-03-01

235

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-?-cyclodextrin (M?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. (paper)

236

Biostimulation of HeLa cells by low-intensity visible light  

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The action of low-intensity red light with lambda=633 nm (a He-Ne laser, a filament lamp with light filters, a dye-laser pumped by a Cu laser) on the intensity of nucleic-acid synthesis in HeLa cells 1.5 hours after irradiation has been studied. It has been shown that the DNA synthesis is stimulated similarly after irradiation both by the He-Ne laser and by an ordinary source. The RNA synthesis intensity does not alter in both cases. A high-repetition-rate radiation at lambda=633 nm acts in the opposite manner: it stimulates the RNA synthesis and the DNA synthesis remains constant. The action spectra of the DNA and RNA synthesis stimulation by continuous light in the range 570/693) nm are presented.

Karu, T.I. (AN SSSR Troitzk. Laser Technology Centre); Kalendo, G.S. (Akademiya Meditsinskikh Nauk SSSR, Moscow. Onkologicheskij Nauchnyj Tsentr); Letokhov, V.S.; Lobko, V.V. (AN SSSR, Moscow. Inst. Spektroskopii)

237

Interaction and cytotoxic effects of hydrophobized chitosan nanoparticles on MDA-MB-231, HeLa and Arpe-19 cell lines.  

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In this work, we investigate the effect of chitosan hydrophobization on the internalization and cytotoxic effect of chitosan-based nanoparticles (NPs) on breast cancer cells (MDA-MB-231), cervical cancer cells (HeLa) and noncancer cells (Arpe-19). We also analyzed the interaction of NPs with a phospholipid (DPPC) membrane model at the airwater interface. An alkylation procedure to insert 8 carbon chains along the chitosan macromolecule with final 10 and 30 % substitution degrees was used. Nuclear magnetic resonance (NMR) and infrared spectroscopes (IR) were used to evaluate the success and extent of the hydrophobization procedure. Size, shape, and charge of NPs were evaluated by dynamic light scattering (DLS), atomic force microscope (AFM), and zeta potential, respectively. The effect of hydrophobicity on NPs was the reduction of the NPs average size, the formation of slightly elongated structures and the enhancing of the interaction of NPs with a DPPC monolayer at the air-water interface. By using fluorescence images on fluorescein-chitosan NPs, we observed a higher internalization of hydrophobic chitosan NPs in cancer cells in comparison with a low internalization of these NPs in normal cells. Even when non modified chitosan NPs were highly internalized in all cell lines, hydrophobized chitosan NPs showed a significantly higher cytotoxic effect on cancer cells in comparison with a lower effect showed by non-modified chitosan NPs on these cells. The cytotoxic effect on the normal cell line used was low for native chitosan NPs and negligible for hydrophobized chitosan NPs. PMID:24444157

Almada, Mario; Burboa, María G; Robles, Emmanuel; Gutiérrez, Luis E; Valdés, Miguel A; Juárez, Josué

2014-01-01

238

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

239

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/?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 ˜ 28% reduction of 12C6+ ion dose for Glu-AuNP treated HeLa cells as compared to control HeLa cells.

Kaur, Harminder; Avasthi, D. K.; Pujari, Geetanjali; Sarma, Asitikantha

2013-07-01

240

Brucella abortus efp gene is required for an efficient internalization in HeLa cells.  

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Numerous chromosomal virulence genes (chv) have been shown to play an important role in the ability of Agrobacterium tumefaciens to transform plants. The A. tumefaciens chvH gene encodes a protein similar in sequence to the Escherichia coli elongation factor P (EF-P). In A. tumefaciens this factor is required for tumor formation and for full expression of the vir genes, exerting its activity at a post-transcriptional level. Cross-complementation assays suggest that the chvH gene and the efp gene of E. coli are functionally homologous. We have cloned and characterized the efp homolog gene in Brucella abortus which has 45% identity to A. tumefaciens chvH and 35% identity to E. coli efp. The gene complemented detergent sensitivity and virulence in the chvH A. tumefaciens mutant, suggesting that both genes are functionally homologous; the growth rate in complex medium also increased to wild type levels. An efp mutant in B. abortus 2308 grew slower in complex media and showed more sensitivity to detergents. Infection assays in J774 macrophage like cells revealed no significant differences between the wild type and the efp mutant strains. The recovery of this mutant from spleens of inoculated mice was equivalent compared to that of the parental strain suggesting that B. abortus efp is not required for virulence in an animal model. However the efp mutant revealed significant differences at 1 h-4 h post-infection in HeLa infection assays compared to the wild type strain, indicating that cellular internalization was affected in non-professional phagocytes. Double immunofluorescence assays for detecting extracellular and intracellular bacteria, demonstrated that the mutant attaches to HeLa cells as the wild type but is deficient in the internalization process, thus indicating that efp is involved in the penetration of Brucella in non-professional phagocytes. PMID:21983596

Iannino, Florencia; Ugalde, Juan E; Iñón de Iannino, Nora

2012-01-01

 
 
 
 
241

The invasion of tobacco mosaic virus RNA induces endoplasmic reticulum stress-related autophagy in HeLa cells.  

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The ability of human cells to defend against viruses originating from distant species has long been ignored. Owing to the pressure of natural evolution and human exploration, some of these viruses may be able to invade human beings. If their 'fresh' host had no defences, the viruses could cause a serious pandemic, as seen with HIV, SARS (severe acute respiratory syndrome) and avian influenza virus that originated from chimpanzees, the common palm civet and birds, respectively. It is unknown whether the human immune system could tolerate invasion with a plant virus. To model such an alien virus invasion, we chose TMV (tobacco mosaic virus) and used human epithelial carcinoma cells (HeLa cells) as its 'fresh' host. We established a reliable system for transfecting TMV-RNA into HeLa cells and found that TMV-RNA triggered autophagy in HeLa cells as shown by the appearance of autophagic vacuoles, the conversion of LC3-I (light chain protein 3-I) to LC3-II, the up-regulated expression of Beclin1 and the accumulation of TMV protein on autophagosomal membranes. We observed suspected TMV virions in HeLa cells by TEM (transmission electron microscopy). Furthermore, we found that TMV-RNA was translated into CP (coat protein) in the ER (endoplasmic reticulum) and that TMV-positive RNA translocated from the cytoplasm to the nucleolus. Finally, we detected greatly increased expression of GRP78 (78 kDa glucose-regulated protein), a typical marker of ERS (ER stress) and found that the formation of autophagosomes was closely related to the expanded ER membrane. Taken together, our data indicate that HeLa cells used ERS and ERS-related autophagy to defend against TMV-RNA. PMID:21729006

Li, Li; Wang, Li; Xiao, Ruijing; Zhu, Guoguo; Li, Yan; Liu, Changxuan; Yang, Ru; Tang, Zhiqing; Li, Jie; Huang, Wei; Chen, Lang; Zheng, Xiaoling; He, Yuling; Tan, Jinquan

2012-04-01

242

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

243

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

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

Bazowski Jessa

2010-10-01

244

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

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

Abdalla A. El Shereef

2011-01-01

245

Growth and cell kinetic changes of HeLa S-3 spheroids following hyperfractionated irradiation  

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Optimal design of the hyperfractionated radiotherapy requires basic radiobiological data such as the critical dose per fraction, number of fractions per day and total equivalent dose, to name a few. As a prelude to in vivo hyperfractionated irradiation, the authors carried out experiments to determine quantitative changes in the proliferation and cell kinetic parameters of multicellular spheroids after hyperfractionated irradiation. Experiments were carried out with HeLa S-3 spheroid growing in MEM culture media. Hyperfractionated irradiation schedules were 1.5 Gy/f, 2f/day and 1.0 Gy/f, 3 f/day. At intervals after irradiation, cell numbers, growth delays and cell cycle distribution of spheroids were determined. The kinetic data were obtained by the use of flow cytometry. The most pronounced changes in cell kinetic parameters were early G/sub 2/M block, proportional to single radiation dose up to 4.0 Gy. There was a corresponding depletion of G1 population as a function of time and dose. In contrast, the maximum accumulation of G/sub 2/M block was delayed as the dose per fraction decreased. The magnitude of the peak accumulation of G/sub 2/M following hyperfractionated irradiation was similar with that of single dose irradiation. Studies are in progress to compare the changes in cell number and growth delays with the kinetic data

246

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

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

247

Reducing the radiation-induced G2 delay causes HeLa cells to undergo apoptosis instead of mitotic death  

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Cells exposed to radiation may undergo death through apoptosis or mitotic death. HeLa cells predominantly undergo mitotic death after irradiation. Treatment of these cells with caffeine has been shown to shorten the G2 delay after irradiation, and to decrease their survival. The kinase inhibitor staurosporine also decreases the radiation-induced G2 delay in HeLa cells. Here we extend these findings to show that the decrease in radiation-induced G2 delay mediated by caffeine or staurosporine is accompanied by a shift in the pathway of cell death from mitotic death to apoptotic death. The increase in apoptosis is further accompanied by decreased clonogenic survival after irradiation. Based on these findings we propose the hypothesis that one mechanism of enhancing cell killing by radiation is to trigger apoptosis by decreasing the G2 delay induced by irradiation. (Author)

248

Hormesis and adaptive response of survival in Hela cells induced by low dose X-ray irradiation  

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The survival fraction in HeLa cells irradiated by low dose X-rays was observed using clone method. The results showed that the survival fraction in the cells irradiated by less than 0.5 Gy X-rays was higher than control, 'hormesis' of HeLa cell survival was obtained and was significant at doses near 0.25 Gy; also, the damage degree of cells induced by the following irradiation was reduced because of pre-treating the cells with low dose D1 of 0.05, 0.75 Gy; it was found from above that 'adaptive response' of cell survival was induced by the low dose irradiation

249

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

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Thiadiazoles are one of the most widely utilized agents in medicinal chemistry, having a wide range of pharmacologic activity. Microtubules (MTs) have always remained a sought-after target in rapidly proliferating cancer cells. We screened for the growth inhibitory effect of synthetic 5-(3-indolyl)-2-substituted-1,3,4-thiadiazoles on cancer cells and identified NMK-TD-100, as the most potent agent. Cell viability experiments using human cervical carcinoma cell line (HeLa cells) indicated that the IC50 value was 1.42±0.11 µM for NMK-TD-100 for 48 h treatment. In further study, we examined the mode of interaction of NMK-TD-100 with tubulin and unraveled the cellular mechanism responsible for its anti-tumor activity. NMK-TD-100 induced arrest in mitotic phase of cell cycle, caused decline in mitochondrial membrane potential and induced apoptosis in HeLa cells. Immunofluorescence studies using an anti-?-tubulin antibody showed a significant depolymerization of the interphase microtubule network and spindle microtubule in HeLa cells in a concentration-dependent manner. However, the cytotoxicity of NMK-TD-100 towards human peripheral blood mononuclear cells (PBMC) was lower compared to that in cancer cells. Polymerization of tissue purified tubulin into microtubules was inhibited by NMK-TD-100 with an IC50 value of 17.5±0.35 µM. The binding of NMK-TD-100 with tubulin was studied using NMK-TD-100 fluorescence enhancement and intrinsic tryptophan fluorescence of tubulin. The stoichiometry of NMK-TD-100 binding to tubulin is 1:1 (molar ratio) with a dissociation constant of ~1 µM. Fluorescence spectroscopic and molecular modeling data showed that NMK-TD-100 binds to tubulin at a site which is very near to the colchicine binding site. The binding of NMK-TD-100 to tubulin was estimated to be ~10 times faster than that of colchicine. The results indicated that NMK-TD-100 exerted anti-proliferative activity by disrupting microtubule functions through tubulin binding and provided insights into its potential of being a chemotherapeutic agent. PMID:24116100

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

2013-01-01

250

Localization of intracellular Ca2+ stores in HeLa cells during infection with Chlamydia trachomatis.  

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Chlamydia trachomatis elementary bodies (EBs) enter epithelial cells within membrane-bound endosomes that aggregate with each other in a calcium-regulated process, but avoid fusion with lysosomes. Annexin III but not I translocates to chlamydial aggregates and inclusions. In this study, we localize the intracellular Ca2+ stores during the course of infection by analyzing the distribution of three intracellular Ca2+ store proteins: calreticulin, type-1 inositol-1,4, 5-trisphosphate receptor (IP3-R), and Sarcoplasmic/Endoplasmic Reticulum Ca2+ ATPase type 2 (SERCA2) in HeLa cells infected with C. trachomatis serovar L2. In uninfected cells, immunofluorescence staining of the proteins showed a fine granular distributed pattern for all three proteins. After infection with C. trachomatis, calreticulin was found at the periphery of chlamydial aggregates and inclusions from 3 to 48 hours post-infection. In infected cells, SERCA2 was intimately associated with chlamydial inclusions after 3 and 24 hours, but not after 48 hours. Moreover, IP3-R was translocated to and colocalized with EB aggregates and chlamydial inclusions and had a distribution very similar to that of SERCA 2. After 24 hours incubation with chlamydiae, there was a local accumulation of [Ca2+]i (105+/-17 nM) in the proximity of chlamydial inclusions, compared to 50+/-13 nM in other parts of the cell cytoplasm. In the absence of extracellular Ca2+, this local accumulation of Ca2+ increased to 295+/-50 nM after adding 50 microM ATP, and to a similar extent after adding 100 nM thapsigargin (Tg). These data indicate that during infection of HeLa cells with chlamydiae, intracellular Ca2+ stores are redistributed, causing local accumulation of Ca2+ in the vicinity of chlamydial inclusions. These changes may trigger the association of certain proteins such as annexins with chlamydia-containing vesicles, and thereby regulation of membrane-membrane interaction during endosome aggregation and inclusion formation. PMID:9841902

Majeed, M; Krause, K H; Clark, R A; Kihlström, E; Stendahl, O

1999-01-01

251

N-methylation of the heterogeneous nuclear ribonucleoproteins in HeLa cells  

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Several of the core proteins on the 40S heterogeneous nuclear ribonucleoprotein particles (hnRNP) from HeLa cells contain N/sup G/,N/sup G/-dimethyl-L-arginine (uDMA). 3-deazaadenosine (c3Ado), an inhibitor of and substrate for s-adenosyl-L-homocysteine hydrolase, has been used to study the methylation patterns of the individual polypeptides. Trimethyllysine and uDMA formation in total cellular protein were inhibited in the presence of the drug while other methylated basic amino acids were unaffected. This inhibition was reversed within 60 min after removal of the drug from the medium. Monolayer HeLa cultures were incubated with [methyl-3H]-L-methoinine for 12 hours in the presence of 50 uM c3Ado. Purified particles were obtained by centrifugation of nuclear extracts on sucrose density gradients. The core proteins were isolated by two-dimensional gel electrophoresis, acid hydrolyzed and analyzed for radioactivity incorporated into methionine and methylated basic amino acids. The ratio of radioactivity incorporated into uDMA relative to that into methionine for the two major particle proteins with molecular weights of 31,000 (A1) and 43,000 (A2) was about 2.0 and 0.2 in control cultures. In the presence of c3Ado, these ratios were depressed 60 to 80%. Results of pulse-chase experiments suggested that A1 and A2 are metabolically stable proteins (t/sub 0.5/ > 75 hr), whether or not the proteins were undermethylated. Monomethyl-L-arginine may be a precursor in the formation of u-DMA

252

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

253

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)

254

NF-?B plays a key role in microcystin-RR-induced HeLa cell proliferation and apoptosis.  

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Microcystins (MCs) are well-known cyanobacterial toxins produced in eutrophic waters and can act as potential carcinogens and have caused serious risk to human health. However, pleiotropic even paradoxical actions of cells exposure to MCs have been reported, and the mechanisms of MC-induced tumorigenesis and apoptosis are still unknown. In this study, we performed the first comprehensive in vitro investigation on carcinogenesis associated with nuclear factor kappa B (NF-?B) and its downstream genes in HeLa cells (Human cervix adenocarcinoma cell line from epithelial cells) exposure to MC-RR. HeLa cells were treated with 0, 20, 40, 60, and 80 µg/mL MC-RR for 4, 8, 12, and 24 h. HeLa cells presented dualistic responses to different doses of MCs. CCK8 assay showed that MC-RR exposure evidently enhanced cell viability of HeLa cells at lower MCs doses. Cell cycle and apoptosis analysis revealed that lower MCs doses promoted G1/S transition and cell proliferation while higher doses of MCs induced apoptosis, with a dose-dependent manner. Electrophoretic mobility shift assay (EMSA) revealed that MC-RR could increase/decrease NF-?B activity at lower/higher MC-RR doses, respectively. Furthermore, the expression of NF-?B downstream target genes including c-FLIP, cyclinD1, c-myc, and c-IAP2 showed the same variation trend as NF-?B activity both at mRNA and protein levels, which were induced by lower doses of MC-RR and suppressed by higher doses. Our data verified for the first time that NF-?B pathway may mediate MC-induced cell proliferation and apoptosis and provided a better understanding of the molecular mechanism for potential carcinogenicity of MC-RR. PMID:24932741

Chen, Liang; Zhang, Xin; Chen, Jun; Zhang, Xuezhen; Fan, Huihui; Li, Shangchun; Xie, Ping

2014-09-01

255

The role of ligand coordination on the cytotoxicity of cationic quantum dots in HeLa cells  

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The effect of ligand structure on the cytotoxicity of cationic CdSe/ZnS quantum dots (QDs) was systematically investigated using mono- and bidentate ligands. Monothiol-functionalized QDs are more cytotoxic than dithiol-functionalized QDs.The effect of ligand structure on the cytotoxicity of cationic CdSe/ZnS quantum dots (QDs) was systematically investigated using mono- and bidentate ligands. Monothiol-functionalized QDs are more cytotoxic than dithiol-functionalized QDs. Electronic supplementary information (ESI) available: Materials, instruments, preparation of cell culture, mass spectrometry, syntheses of surface ligands and cationic QDs, MALDI-MS spectra of QDs, cellular uptake and cytotoxicity of additional QDs after incubation with HeLa cells for 24 h, cell viability after QD incubation with HeLa cells for 24 h in the absence and presence of NAC, and necrosis-mediated cell death experiments. See DOI: 10.1039/c3nr04037b

Yeh, Yi-Cheun; Saha, Krishnendu; Yan, Bo; Miranda, Oscar R.; Yu, Xi; Rotello, Vincent M.

2013-11-01

256

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)

257

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

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

Hellen Rotinberg

2008-12-01

258

Uptake of [3H]ouabain from the cell surface into the lysosomal compartment of HeLa cells  

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

259

Human cytosolic thymidine kinase: purification and physical characterization of the enzyme from HeLa cells  

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The mammalian cytosolic thymidine kinase is one of a number of enzymes involved in DNA replication whose activities increase dramatically during S phase of the cell cycle. As a first step in defining the mechanisms that control the S phase induction of thymidine kinase activity, the authors have purified the human enzyme from HeLa cells and raised a specific immune serum against the purified protein. The enzyme was isolated from cells arrested in S phase by treatment with methotrexate and purified to near homogeneity by ion-exchange and affinity chromatography. Stabilization of the purified enzyme was achieved by the addition of digitonin. An electrophoretic R/sub m/ of 0.2 in nondenaturing gels characterizes the purified enzyme activity as cytosolic thymidine kinase. The enzyme has a Stoke's radius of 40 A determined by gel filtration and a sedimentation coefficient of 5.5 S determined by glycerol gradient sedimentation. Based on these hydrodynamic values, a native molecular weight of 96,000 was calculated for the purified enzyme. When electrophoresed in denaturing sodium dodecyl sulfate-polyacrylamide gels under reducing conditions, the most purified enzyme fraction was found to contain one predominant polypeptide of M/sub r/ = 24,000. Several lines of evidence indicate that this polypeptide is responsible for thymidine kinase enzymatic activity

260

A New Diterpene from Litsea cubeba Fruits: Structure Elucidation and Capability to Induce Apoptosis in HeLa Cells  

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Full Text Available A new diterpene, identified as (+-6-(4-hydroxy-4-methyl-2-pentenoyl-4,6-dimethyl-5-(3-methyl-2-butenyl-1,3-cyclohexadienecarbaldehyde (1, cubelin, was isolated from a methanol extract of Litsea cubeba fruits by normal phase column chromatography and purified by preparative HPLC. The structure elucidation was conducted by spectroscopic methods (UV, IR, ESI-TOF-MS, 1-D and 2-D NMR. Cubelin exhibited activity against HeLa cell viability and proliferation. The cells also exhibited changes in nuclear morphology which are hallmarks of apoptotic cell death. The presence of cleaved caspase-3/-7, caspase-8 and caspase-9 in the cubelin treated population indicated the potential of the compound to induce apoptosis in HeLa cells via both intrinsic and extrinsic pathways.

Piyapat Trisonthi

2014-05-01

 
 
 
 
261

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

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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=26 mW; lambda=.670 nm) 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. PMID:15002747

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

2003-12-01

262

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

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

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

2003-12-01

263

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

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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

264

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

Scientific Electronic Library Online (English)

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

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

265

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

266

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

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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.

267

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 D0 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 G2-phase delay. Addition of polyamines to cultures for short periods prior to irradiation restored normal radioresistence and reversed the anomalous features of the G2-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 G2-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 G2-phase delay response may be the determining factors. 34 refs., 8 figs., 3 tabs

268

Permeability and gating properties of human connexins 26 and 30 expressed in HeLa cells.  

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Human connexins 26 and 30 were expressed either through the bicistronic pIRES-EGFP expression vector or as EYFP-tagged chimeras. When transiently transfected in communication-incompetent HeLa cells, hCx26-pIRES transfectants were permeable to dyes up to 622 Da, but were significantly less permeable to 759 Da molecules. Under the same conditions, permeability of hCx26-EYFP fusion products was comparable to that of hCx26-pIRES, but with significant increase in diffusion at 759 Da, possibly as a consequence of having selected large fluorescent junctional plaques. Dye transfer was limited to 457 Da in hCx30-EYFP transfectants. When reconstructed from confocal serial sections, fluorescent plaques formed by hCx26-EYFP and hCx30-EYFP appeared irregular, often with long protrusions or deep invagination. Similar plaques were observed following immunostaining both in cells transfected with hCx26-pIRES and in HeLa cells stably transfected with mouse Cx26. Tissue conductance (Tg(j)) displayed significantly smaller values (28.8+/-1.8 nS) for stably transfected mCx26 than transiently transfected hCx26 (43.5+/-3.3 nS). These differences reflected in distinct functional dependence of normalized junctional conductance (G(j)) on transjunctional voltage (V(j)). The half-activation voltage for G(j) was close to +/-95 and +/-58 mV in mCx26 and hCx26, respectively. The corresponding parameters for hCx30 transfectants were Tg(j)= 45.2 +/- 3.5 nS and V(0)= +/- 34 mV. These results highlight unexpected differences between mCx26 and hCx26 in this expression system, reinforce the concept that channel permeability may be related to Cx level expression, and indicate that fusion of hCx30 to GFP colour mutants produces channels that are suitable for permeability and gating studies. PMID:12767933

Beltramello, Martina; Bicego, Massimiliano; Piazza, Valeria; Ciubotaru, Catalin D; Mammano, Fabio; D'Andrea, Paola

2003-06-13

269

Conductive and kinetic properties of connexin45 hemichannels expressed in transfected HeLa cells.  

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Human HeLa cells transfected with mouse connexin Cx45 were used to examine the conductive and kinetic properties of Cx45 hemichannels. The experiments were carried out on single cells using a voltage-clamp method. Lowering the [Ca2+]o revealed an extra current. Its sensitivity to extracellular Ca2+ and gap junction channel blockers (18alpha-glycyrrhetinic acid, palmitoleic acid, heptanol), and its absence in non-transfected HeLa cells suggested that it is carried by Cx45 hemichannels. The conductive and kinetic properties of this current, Ihc, were determined adopting a biphasic pulse protocol. Ihc activated at positive Vm and deactivated partially at negative Vm. The analysis of the instantaneous Ihc yielded a linear function ghc,inst = f(Vm) with a hint of a negative slope (ghc,inst: instantaneous conductance). The analysis of the steady-state Ihc revealed a sigmoidal function ghc,ss=f(Vm) best described with the Boltzmann equation: Vm,0= -1.08 mV, ghc,min=0.08 (ghc,ss: steady-state conductance; Vm,0: Vm at which ghc,ss is half-maximally activated; ghc,min: minimal conductance; major charge carriers: K+ and Cl-). The ghc was minimal at negative Vm and maximal at positive Vm. This suggests that Cx45 connexons integrated in gap junction channels are gating with negative voltage. Ihc deactivated exponentially with time, giving rise to single time constants, taud. The function taud = f(Vm) was exponential and increased with positive Vm (taud=7.6 s at Vm=0 mV). The activation of Ihc followed the sum of two exponentials giving rise to the time constants, taua1 and taua2. The function taua1=f(Vm) and taua2 = f(Vm) were bell-shaped and yielded a maximum of congruent with 0.6 s at Vm congruent with -20 mV and congruent with 4.9 s at Vm congruent with 15 mV, respectively. Neither taua1 =f(Vm) nor taua2 = f(Vm) coincided with taud=f(Vm). These findings conflict with the notion that activation and deactivation follow a simple reversible reaction scheme governed by first-order voltage-dependent processes. PMID:15457371

Bader, P; Weingart, R

2004-06-01

270

Compositional analysis of glucosaminyl(acyl)phosphatidylinositol accumulated in HeLa S3 cells.  

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GlcN(acyl)PtdIns, a derivative of phosphatidylinositol (PtdIns) in which glucosamine and a fatty acid are linked to inositol hydroxyl groups, has been proposed to be an intermediate in the mammalian biosynthetic pathway for glycosylphosphatidylinositol (glycosyl-PtdIns) anchors of membrane proteins. In this report, GlcN(acyl)PtdIns metabolically labeled with [3H]inositol is shown to accumulate in a HeLa S3 cell subline. The amount of GlcN(acyl)PtdIns in these HeLa S3 cells is about 10(7) molecules/cell, a level comparable to those of the most abundant glycosyl-PtdIns-containing molecules reported to date. GlcN(acyl)PtdIns was purified by a two-step procedure involving octyl-Sepharose and thin-layer chromatography. Octyl-Sepharose separated phospholipids according to their number of hydrocarbon chains: one in 2-lysoPtdIns, two in PtdIns, and three in GlcN(acyl)PtdIns. Purification also was aided by prior treatment of lipid extracts with bee venom phospholipase A2, an enzyme that did not cleave GlcN(acyl)PtdIns. The GlcN-inositol head group in purified GlcN(acyl)PtdIns was confirmed by a number of procedures, including cation-exchange chromatography and mass spectrometry; after radiomethylation, an equal molar ratio of GlcN(Me)2/inositol was measured. Fatty acid analysis indicated an overall stoichiometry of 2.3 mol fatty acid/mol inositol with palmitic (16:0), stearic (18:0) and oleic (18:1) acids being predominant. Analysis of GlcN(acyl)inositol produced by HF fragmentation showed that palmitate was the acyl group attached to inositol and indicated that stearic and oleic acids were in the glycerolipid. Base methanolysis revealed that about 15% of the purified GlcN(acyl)PtdIns contained alkylglycerol. A substantial conversion of GlcN(acyl)PtdIns to a slightly more polar lipid occurred after overnight incubation in even mildly alkaline buffers. Although the current data do not allow proposal of a structure for this lipid, its formation from GlcN(acyl)PtdIns may be important because the conversion appeared to occur in vivo. PMID:7588771

Sevlever, D; Humphrey, D R; Rosenberry, T L

1995-10-01

271

Quantitative and real-time effects of carbon quantum dots on single living HeLa cell membrane permeability  

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The interaction between carbon quantum dots (CQDs) and a single living cell was explored in real time. Here, we provide the quantitative data on the permeability of the HeLa cell membrane in the presence of CQDs with different surface functional groups (CQDs terminated with -OH/-COOH (CQD-OH), -PEG (CQD-PEG), and -NH2 (CQD-NH2)). Although these CQDs have very low toxicity towards HeLa cells, they still increase the cell membrane permeability by 8%, 13%, and 19% for CQD-PEG, CQD-OH, and CQD-NH2, respectively, and this kind of permeability was irreversible. These observations are valuable for promoting the bio-applications of carbon nanostructures in living systems.The interaction between carbon quantum dots (CQDs) and a single living cell was explored in real time. Here, we provide the quantitative data on the permeability of the HeLa cell membrane in the presence of CQDs with different surface functional groups (CQDs terminated with -OH/-COOH (CQD-OH), -PEG (CQD-PEG), and -NH2 (CQD-NH2)). Although these CQDs have very low toxicity towards HeLa cells, they still increase the cell membrane permeability by 8%, 13%, and 19% for CQD-PEG, CQD-OH, and CQD-NH2, respectively, and this kind of permeability was irreversible. These observations are valuable for promoting the bio-applications of carbon nanostructures in living systems. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr06590a

Kong, Weiqian; Liu, Juan; Liu, Ruihua; Li, Hao; Liu, Yang; Huang, Hui; Li, Kunyang; Liu, Jian; Lee, Shuit-Tong; Kang, Zhenhui

2014-04-01

272

Cytotoxicity of berberine on human cervical carcinoma HeLa cells through mitochondria, death receptor and MAPK pathways, and in-silico drug-target prediction.  

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Berberine, a natural product, has been widely used to treat hyperlipoidemia and intestinal diseases. In the present paper, berberine showed a significant anti-proliferative effect to human cervical carcinoma HeLa cells confirmed by 3-(4,5)-dimethyl-thiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide (MTT), flow cytometry analysis (FCM) and so on. The methods including western blotting, radioimmunity assay (RIA), reverse transcription-polymerase chain reaction (RT-PCR) were used to investigate protein and mRNA expressions. We found that Bcl-2/Bax ratio was significantly decreased and cytochrome c was released from mitochondrion to cytosol, which indicated that the mitochondrial pathway was activated by berberine. The up-regulation of Fas, FasL, TNF-alpha and TRAF-1 indicated the involvement of the death receptor pathway in the process of berberine-induced apoptosis. Furthermore caspase-3 and caspase-8 were activated as a central event of apoptosis, and the levels of phosphorylation of mitogen-activated protein kinases (MAPKs) were also investigated. In addition, the increased expression of p53 was also observed in berberine-treated HeLa cells, and as a node point of these different pathways in a protein-protein interaction network constructed by GeneGo software, p53 might be the possible drug-target of berberine's anti-cancer on HeLa cells, which was predicted by a flexible ligand-protein inverse docking program, INVDOCK. This study is benefit for clarifying the mechanism of berberine's anti-tumor effect and might be helpful to find therapy-target for treatment of human cervical carcinoma. PMID:20656010

Lu, Binan; Hu, Mingming; Liu, Kexin; Peng, Jinyong

2010-09-01

273

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

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

Wei Zhu

2013-03-01

274

Viscum album L. Extracts Protects HeLa Cells against Nuclear and Mitochondrial DNA Damage.  

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Viscum album L. is a semiparasitic plant grown on trees and widely used for the treatment of many diseases in traditional and complementary therapy. It is well known that some activities of Viscum album extracts are varied depending on the host trees, such as antioxidant, apoptosis-inducing, anticancer activities of the plant. The aim of the present study is to examine the comparative effects of methanolic extracts of V. album grown on three different host trees (locust tree, lime tree, and hedge maple tree) on H(2)O(2)-induced DNA damage in HeLa cells. Oxidative damage in mitochondrial DNA and two nuclear regions was assessed by QPCR assay. The cells were pretreated with methanolic extracts (10??g/mL) for 48?h, followed by the treatment with 750??M H(2)O(2) for 1 hour. DNA damage was significantly induced by H(2)O(2) while it was inhibited by V. album extracts. All extracts completely protected against nuclear DNA damage. While the extract from lime tree or white locust tree entirely inhibited mitochondrial DNA damage, that from hedge maple tree inhibited by only 50%. These results suggest that methanolic extracts of V. album can prevent oxidative DNA damage, and the activity is dependent on the host tree. PMID:22988477

Onay-Uçar, Evren; Erol, Ozlem; Kandemir, Ba?ak; Merto?lu, Elif; Karagöz, Ali; Arda, Nazl?

2012-01-01

275

Antioxidant potential of Ulva rigida extracts: protection of HeLa cells against H?O? cytotoxicity.  

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The rising demand for natural antioxidants instead of synthetic materials, especially in biomedical applications, has led to increased interest in the search for bioactive compounds with potent antioxidant activity. In the present study, we tested the antioxidant effect of both a crude extract and an ethanol precipitate of Ulva rigida in HeLa cells exposed to hydrogen peroxide (H?O?). HeLa cells treated with H?O? (1 mmol l?¹ for 3 h) exhibited significant damage to their morphology, a significant decrease in cell survival, and a remarkable leakage of lactate dehydrogenase (LDH). However, the co-exposure of cells to H?O? and the crude extract or ethanol precipitate of U. rigida induced fewer morphological cytotoxic effects, a significant increase in cell viability, and a significant decrease in LDH release. Biochemical studies have demonstrated that U. rigida extracts have a strong radicalscavenging activity and contain protein, sugar, and phenolic content. The overall results suggest that U. rigida extracts protect HeLa cells from death induced by oxidative stress, and it is likely that these effects are related to the phenolic, protein, and polysaccharide compounds contained in this alga. Hence, U. rigida can be used to treat diseases ascribed to oxidative disorders. PMID:24088791

Mezghani, Sana; Bourguiba, Ines; Hfaiedh, Imen; Amri, Mohamed

2013-09-01

276

Investigation of siRNA Nanoparticle Formation Using Mono-Cationic Detergents and Its Use in Gene Silencing in Human HeLa Cells  

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Full Text Available The focus of recent research has been on the development of siRNA vectors to achieve an innovative gene therapy. Most of the conventional vectors are siRNA nanoparticles complexed with cationic polymers and liposomes, making it difficult to release siRNA. In this study, we report on the use of MCD, a quaternary ammonium salt detergent containing a long aliphatic chain (L-chain as an siRNA complexation agent using human HeLa cells (a model cancer cell. We prepared siRNA nanoparticles using various MCDs, and measured the diameters and zeta-potentials of the particles. The use of an MCD with a long L-chain resulted in the formation of a positively charged nanoparticle. In contrast, a negatively charged nanoparticle was formed when a MCD with a short L-chain was used. We next evaluated the gene silencing efficiency of the nanoparticles using HeLa cells expressing the luciferase protein. The results showed that the siRNA/MCD nanoparticles showed a higher gene silencing efficiency than Lipofectamine 2000. We also found that the efficiency of gene silencing is a function of the length of the alkyl chain in MCD and zeta-potential of the siRNA/MCD nanoparticles. Such information provides another viewpoint for designing siRNA vectors.

Hideyoshi Harashima

2013-11-01

277

Direct observation of potassium ions in HeLa cell with ion-selective nano-pipette probe  

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The local concentration of potassium ion in a single HeLa cell was observed with an ion-selective nano-pipette probe. Ion selectivity was achieved by using a polyvinyl chloride film with selected ionophores placed within the nano-pipette. Both alternating and constant bias voltages were applied to the counter electrode for the observation of local ion concentrations with a response time of less than 0.1 s. These measurements were enabled by a low-current detection system prepared specifically for this study. The difference in local potassium concentrations between inside a living HeLa cell and the surrounding solution was approximately 100 mM, while no difference in potassium ion concentration was observed between the interior of dead cells and the surrounding solution.

Takami, Tomohide; Iwata, Futoshi; Yamazaki, Koji; Wan Son, Jong; Lee, Joo-Kyung; Ho Park, Bae; Kawai, Tomoji

2012-02-01

278

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

International Nuclear Information System (INIS)

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

279

Shape of the fluidity gradient in the plasma membrane of living HeLa cells.  

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The shape of the fluidity gradient of the outer hemi-leaflet of the plasma membrane of living HeLa cells was determined using a series of n-(9-anthroyloxy) fatty acid probes where n = 2, 3, 6, 7, 9, 10, 11, 12, and 16. Fluorescence uptake and steady-state anisotropy values were obtained with a flow cytometer capable of continuous recording over time of vertical and horizontal emission intensities, and of the output of these intensities as calculated anisotropy values. The fluorescence uptake of all of the membrane probes was rapid up to about 15 min. The magnitudes of the uptake of fluorescence were, for the n-(9-anthroyloxy) series, in the order 2 greater than 3 greater than 6 greater than 7 greater than 9 greater than 10 greater than 11 = 12 = 16. Anisotropy values were constant from 5 to 30 min after addition of the various probes, and the magnitudes were in the order 7 greater than 6 greater than 9 = 10 greater than 2 = 3 greater than 11 greater than 12 greater than 16, indicative of the shape of the fluidity gradient. No differences were noted between the values obtained with 12-(9-anthroyloxy) stearic acid and 12- (9-anthroyloxy) oleate. The kinetics of anisotropy exhibited by those probes with the anthroyloxy group in positions deeper than 9, where initially higher values declined until equilibrium was reached, were probably indicative of an energy barrier at the approximate depth sensed by 7 AS. PMID:2324646

Collins, J M; Dominey, R N; Grogan, W M

1990-02-01

280

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

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

Halder Rashi

2012-03-01

 
 
 
 
281

Role of FeCl3 in 67Ga uptake by HeLa S3 cells in vitro  

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The effect of FeCl3 on 67Ga uptake by HeLa S3 cells was studied in vitro. Uptake of 67Ga by HeLa S3 cells was influenced by the concentration of serum in the medium and especially that of transferrin added to the medium. Therefore, it is assumed that a mechanism of 67Ga uptake exists mediated by transferrin. However, when 10-2-10-1 mM FeCl3 was added to the medium in the same experimental system, a remarkable increase of 67Ga uptake was noted. Results of equilibrium dialysis, showed that binding of 67Ga to transferrin was inhibited by the administration of FeCl3. Moreover, the formation of a complex of 67Ga and FeCl3, which did not pass through the cellulose membrane, was observed. A correlation was found between an increase in 67Ga uptake by HeLa S3 cells and the formation of the 67Ga-Fe complex. These results indicate that another mechanism of 67Ga accumulation, namely the formation of a 67Ga-Fe complex plays a role in the uptake of 67Ga by tumor cells in the presence of iron. (orig.)

282

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

283

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

284

Squamous cell skin cancer  

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... of skin cancer. Other common types of skin cancer are: Basal cell Melanoma ... Squamous cell cancer spreads faster than basal cell cancer , but still may grow slowly. It may spread to other parts of the body, including internal organs.

285

Cytotoxic isolates of Helicobacter pylori from Peptic Ulcer Diseases decrease K+-dependent ATPase Activity in HeLa cells  

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Full Text Available Abstract Background Helicobacter pylori is a Gram negative bacterium that plays a central role in the etiology of chronic gastritis and peptic ulcer diseases. However, not all H. pylori positive cases develop advanced disease. This discriminatory behavior has been attributed to the difference in virulence of the bacteria. Among all virulence factors, cytotoxin released by H. pylori is the most important factor. In this work, we studied variation in H. pylori isolates from Indian dyspeptic patients on the basis of cytotoxin production and associated changes in K+-dependent ATPase (one of its targets enzyme activity in HeLa cells. Methods The patients were retrospectively grouped on the basis of endoscopic and histopathological observation as having gastritis or peptic ulcer. The HeLa cells were incubated with the broth culture filtrates (BCFs of H. pylori isolates from patients of both groups and observed for the cytopathic effects: morphological changes and viability. In addition, the K+-dependent ATPase activity was measured in HeLa cells extracts. Results The cytotoxin production was observed in 3/7 (gastritis and 4/4 (peptic ulcer H. pylori isolates. The BCFs of cytotoxin producing H. pylori strains reduced the ATPase activity of HeLa cells to 40% of that measured with non-cytotoxin producing H. pylori strains (1.33 ?mole Pi/mg protein and 3.36 ?mole Pi/mg protein, respectively, p Conclusions Our results suggest that the isolation of cytotoxic H. pylori is more common in severe form of acid peptic diseases (peptic ulcer than in gastritis patients from India. Also the cytotoxin released by H. pylori impairs the ion-transporting ATPase and is a measure of cytotoxicity.

Archana Ayyagari

2003-11-01

286

Immunofluorescence method for detection of Ebola virus immunoglobulin g, using HeLa cells which express recombinant nucleoprotein.  

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A novel recombinant baculovirus which expresses Ebola virus (EBO) nucleoprotein (NP) under the control of the cytomegalovirus immediate-early promoter was constructed. HeLa cells abortively infected with the baculovirus expressed EBO NP, and this was used as an immunofluorescent (IF) antigen to detect EBO immunoglobulin G (IgG) antibody. This IF method has high efficacy in detecting EBO IgG antibody in clinical specimens, indicating its usefulness in the diagnosis of EBO infections and seroepidemiological studies. PMID:11158150

Saijo, M; Niikura, M; Morikawa, S; Kurane, I

2001-02-01

287

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

288

Selective eradication of cancer cells in vitro  

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A simple system consisting of cultured HeLa (human cancer) and WI38 (normal human fetal lung) cells and the control cultures of the individual cells were set up to test and compare the effects of the cell cycle-active agents /sup 125/I-iododeoxyuridine (/sup 125/IUdR) and hydroxyurea (HU) on cell survival. The presence of cells and growth after treatment were used as a positive indication of survival. The experimental cultures were first seeded with WI38 cells and allowed to grow to confluency before adding 1.0 x 10/sup 5/ HeLa cells. After two days of treatment-free growth, the co-cultures were continuously treated with /sup 125/IUdR (0.5-2.0 ?Ci/ml, carrier free) or HU (1.0 x 10/sup -9/ and 1.0 x 10/sup -3/M). At the termination of treatment the co-cultures were split 3 to 1 and incubated for seven days. As expected, there was little or no detectable effect on the growth of WI38 cells treated with HU or /sup 125/IUdR while the cells were confluent. However, HeLa cells were reduced by 1.0 x 10/sup -3/M HU and were eradicated after all concentrations of /sup 125/IUdR

289

DNA of HeLa cells during caffeine-promoted recovery from X-ray induced G2 arrest  

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Progression of X-irradiated HeLa cells from G2 arrest through mitosis was promoted by 1mM caffeine. Caffeine promoted the return from abnormally high levels of radiation-induced immunoreactivity to antinucleoside antibodies, which indicates persistent DNA strand separation, to the low levels normally found in G2. With caffeine, the irradiated cells progressed through mitosis, producing daughter cells with the normal G1 content of DNA. Without caffeine, the DNA content of individual radiation-arrested cells retained G2 values and the abnormally high levels of immunoreactivity to antinucleoside antibodies. (author)

290

Dr fimbriae operon of uropathogenic Escherichia coli mediate microtubule-dependent invasion to the HeLa epithelial cell line.  

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Escherichia coli Dr adhesin and decay-accelerating factor (DAF) receptor-mediated interaction was proposed as the mechanism of ascending urinary tract infection (UTI) and chronic interstitial nephritis. This report provides novel evidence for Dr fimbriae operon-mediated invasive capacity of Dr+ E. coli. Insertional mutants draE, draC, and draB, and adherent draD and UV-inactivated BN406 were unable to enter HeLa cells. Complementation of the dra mutation restored invasiveness. Internalization was inhibited by anti-Dr fimbriae IgG (100%), anti-SCR-3 domain of DAF (75%), and nocodazole (95%). Increased receptor-ligand density occurred at the site of internalization. Internalized Dr+ E. coli did not significantly multiply in the HeLa cell line. Accordingly, the dra operon and DAF were required for microtubule-dependent internalization of E. coli to HeLa cells. The relatively low invasion and multiplication rates of Dr+ E. coli may hypothetically contribute to the postattachment steps of ascending UTI and chronic renal infection. PMID:9207362

Goluszko, P; Popov, V; Selvarangan, R; Nowicki, S; Pham, T; Nowicki, B J

1997-07-01

291

Effects of DNA polymerase inhibitors on replicative and repair DNA synthesis in ultraviolet-irradiated HeLa cells  

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Aphidicolin specifically inhibits eukaryotic DNA polymerase ?, while 2',3'-dideoxythymidine 5'-triphosphate (d2TTP) inhibits DNA polymerase ? and ? but not ?. 1-?-D-Arabinofuranosylcytosine 5'-triphosphate (araCTP) inhibits both DNA polymerase ? and ? although to a different extent. Here we measured the effects of these inhibitors on repair DNA synthesis of U.V.-irradiated HeLa cells by two different methods. Firstly, aphidicolin, 1-?-D-arabinofuranosylcytosine (araC, a precursor of araCTP) and 2',3'-dideoxythimidine (d2Thd, a precursor of d2TTP) were added directly to the culture medium. In this case, aphidicolin and araC strongly inhibited replicative DNA synthesis of HeLa cells, and they also inhibited repair synthesis after U.V.-irradiation but to a much lesser extent. In contrast, high concentrations of d2Thd inhibited repair DNA synthesis to a higher extent than replicative DNA synthesis. Secondly, the active form of inhibitor, d2TTP, was microinjected directly into cytoplasm or nuclei or U.V.-irradiated HeLa cells. Microinjection of d2TTP effectively inhibited repair synthesis. The microinjection of d2TTP, into either cytoplasm or nucleus, strongly inhibited replicative synthesis. These results might indicate that multiple DNA polymerases are involved in repair synthesis as well as in replicative synthesis

292

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

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Lethal toxin of Clostridium sordellii (MLD 150 ng/kg) is one of the most potent Clostridial toxins and is responsible for most of the diseases including sudden death syndrome in cattle, sheep and toxic shock syndrome, necrotizing faciitis, neonatal omphalitis and gangrene in humans. Lethal toxin (TcsL) is a single chain protein of about 270 kDa. In the present study, 1.6 kb DNA fragment encoding for the catalytic domain of TcsL was PCR amplified, cloned in pQE30 UA vector and expressed in E. coli SG 13009. The expression of recombinant lethal toxin protein (rTcsL) was optimized and it was purified under native conditions using a single step Ni-NTA affinity chromatography. The purified recombinant protein was used for the production of polyclonal antibodies in mice and rabbit. The raised antibodies reacted specifically with the purified rTcsL and intact native lethal toxin on Western blot. The biological activity of the recombinant protein was tested in HeLa cells where it showed the cytotoxicity. Further, the polyclonal antibodies were used for in-vitro neutralization of purified rTcsL, acid precipitated C. sordellii and C. difficile native toxins in HeLa cells. Mice and rabbit anti-rTcsL sera effectively neutralized the cytotoxicity of rTcsL and C. sordellii native toxin but it did not neutralize the cytotoxicity of C. difficile toxin in HeLa cells. PMID:22894159

Arya, Preetika; Ponmariappan, S; Singh, Lokendra; Prasad, G B K S

2013-02-01

293

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

294

Laserthermia enhances the clastogenic effects of antineoplastic agents in aerobic and chronically hypoxic HeLa cells in vitro.  

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The interactive clastogenic effects of Nd-YAG laser induced hyperthermia (laserthermia) in combination with antineoplastic agents on normally oxygenated and chronically hypoxic HeLa cells were investigated. Exponentially growing HeLa cells were treated with bleomycin sulfate (BLM) (2-4 microg/ml), adriamycin (ADM) (2-4 microg/ml) and actinomycin D (ACT) (0.2-0.4 microg/ml) alone or in combination with laser at various powers (7-13 W) or different laser induced elevated temperatures (39.5-43.5 degrees C). HeLa cells were incubated with 3 microg/ml cytochlasin B for 36 h after treatments and the frequency of micronuclei (MN) were determined in binucleated cells. Results showed a relatively high frequency of MN formation after drug treatments in normally oxic and chronically hypoxic cells, although there was a decrease in the frequency of MN in hypoxic cells compared to oxygenated cells. Laserthermia at various powers and different induced temperatures produced a slight increase in MN formation both in oxic and hypoxic cells. When drug treatment and laserthermia was combined, a profound synergistic effect in MN formation was observed for all three drugs used in these experiments. ACT at a concentration of ten times lower than ADM and BLM produced similar effect. Also, ADM showed a marked synergistic effect with laserthermia compared to BLM at similar concentrations. This study suggests that laserthermia in combination with ADM, BLM or ACT would have a greater genotoxic effect on hypoxic cell populations. Therefore, Nd-YAG laser induced hyperthermia may be a useful modality for elimination of the radioresistant hypoxic cells. PMID:11323094

Mozdarani, H; Monfared, A S

2001-06-10

295

Analysis of microRNA expression profiles during the cell cycle in synchronized HeLa cells.  

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Cell cycle progression is regulated by both transcriptional and post-transcriptional mechanisms. MicroRNAs (miRNAs) emerge as a new class of small non-coding RNA regulators of cell cycle as recent evidence suggests. It is hypothesized that expression of specific miRNAs oscillates orderly along with cell cycle progression. However, the oscillated expression patterns of many candidate miRNAs have yet to be determined. Here, we describe miRNA expression profiling in double-thymidine synchronized HeLa cells as cell cycle progresses. Twenty-five differentially expressed miRNAs were classified into five groups based on their cell cycle-dependent expression patterns. The cyclic expression of six miRNAs (miR-221, let-7a, miR-21, miR-34a, miR-24, miR-376b) was validated by real-time quantitative RT-PCR (qRT-PCR). These results suggest that specific miRNAs, along with other key factors are required for maintaining and regulating proper cell cycle progression. The study deepens our understanding on cell cycle regulation. [BMB reports 2009; 42(9): 593-598]. PMID:19788861

Zhou, Jue-Yu; Ma, Wen-Li; Liang, Shuang; Zeng, Ye; Shi, Rong; Yu, Hai-Lang; Xiao, Wei-Wei; Zheng, Wen-Ling

2009-09-30

296

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

297

Radiosensitizing effect of 2,4-dinitroimidazole-1-ethanol and its cytotoxicity in HeLa S3 cells  

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Using cultured HeLa S3 cells, the radiosensitizing and cytotoxic effects of newly synthesized derivatives of dinitroimidazole were investigated and compared with those of misonidazole. 2,4-dinitroimidazole-1-ethanol radiosensitized hypoxic cells selectively. At 5 mM misonidazole, the enhancement ratio was 1.95; with 0.5 mM 2,4-dinitroimidazole-1-ethanol, almost the same enhancement could be obtained. This indicates that the radiosensitizing effect of the latter agent was about 10 times greater than that of misonidazole. However, its cytotoxicity was twice that of misonidazole under hypoxic conditions and there was no apparent differential cytotoxicity to hypoxic and aerobic cells. (orig.)

298

Radiosensitizing effect of 2,4-dinitroimidazole-1-ethanol and its cytotoxicity in HeLa S3 cells  

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Using cultured HeLa S3 cells, the radiosensitizing and cytotoxic effects of newly synthesized derivatives of dinitroimidazole were investigated and compared with those of misonidazole. 2,4-dinitroimidazole-1-ethanol radiosensitized hypoxic cells selectively. At 5 mM misonidazole, the enhancement ratio was 1.95; with 0.5 mM 2,4-dinitroimidazole-1-ethanol, almost the same enhancement could be obtained. This indicates that the radiosensitizing effect of the latter agent was about 10 times greater than that of misonidazole. However, its cytotoxicity was twice that of misonidazole under hypoxic conditions and there was no apparent differential cytotoxicity to hypoxic and aerobic cells.

Shibata, C.; Mori, T.; Hori, H.; Inayama, S.

1981-07-01

299

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)

300

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

 
 
 
 
301

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

302

Human small cell lung cancer cell lines express functional atrial natriuretic peptide receptors.  

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Small cell lung cancer cell (SCLC) lines, NCI-H82, NCI-H660, and NCI-H1284, and HeLa cells were analyzed for the presence of atrial natriuretic peptide (ANP) receptors. In these SCLC cell lines and HeLa cells, ANP A receptor mRNA was identified by Southern blot analyses of polymerase chain reaction products and RNase protection assays using poly(A)(+)-selected RNA. Saturable binding assays revealed that HeLa cells had 2000 to 5000 high affinity atrial natriuretic peptide receptors per cell with a dissociation constant of 140 pM. In the SCLC cell lines, the binding was saturable but too low to accurately estimate the number of binding sites. After addition of human ANP, radioimmunoassays revealed accumulation of cyclic GMP in SCLC cells as well as HeLa cells in a dose-dependent fashion. The half-maximal stimulation concentration of cyclic GMP accumulation in HeLa and these SCLC cell lines was approximately 2 nM. Tetrazolyl blue assays and tritiated thymidine incorporation did not show any remarkable growth inhibition or growth stimulation of SCLC cell lines after addition of human ANP up to 3.3 microM, more than 1000-fold greater than the half-maximal stimulation concentration of cyclic GMP accumulation. Our results indicate that human SCLC cells express functional ANP receptors but ANP addition produced no detectable change in their growth pattern. PMID:8391389

Ohsaki, Y; Yang, H K; Le, P T; Jensen, R T; Johnson, B E

1993-07-01

303

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-statone 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

304

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

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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

305

OppA, the ecto-ATPase of Mycoplasma hominis induces ATP release and cell death in HeLa cells  

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Full Text Available Abstract Background In the facultative human pathogen Mycoplasma hominis, which belongs to the cell wall-less Mollicutes, the surface-localised substrate-binding domain OppA of the oligopeptide permease was characterised as the main ecto-ATPase. Results With the idea that extra-cellular ATP could only be provided by the infected host cells we analysed the ATP release of HeLa cells after incubation with different preparations of Mycoplasma hominis: intact bacterial cells, the membrane fraction with or without OppA, recombinant OppA as well as an ATPase-deficient OppA mutant. Release of ATP into the supernatant of the HeLa cells was primarily determined in all samples lacking ecto-ATPase activity of OppA. In the presence of the ATPase inhibitor DIDS the amount of ATP in the OppA-containing samples increased. This increase was maximal after incubation with fractions containing OppA protein indicating that OppA is involved in ATP release and subsequent hydrolysis. Real-time PCR analyses revealed that the proliferation of HeLa cells is reduced after infection with M. hominis and flow cytometry experiments established that OppA induces greater apoptosis than necrosis of HeLa cells whereas the preservation of ecto-ATPase activity of OppA induces apoptosis. Conclusion The OppA induced ATP-release and -hydrolysis induced cell death of M. hominis infected HeLa cells was predominantly due to apoptosis rather than necrosis. Future work will elucidate whether the induction of apoptosis is indispensable for survival of these non-invasive pathogen.

Henrich Birgit

2008-04-01

306

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 [32P]NAD, chromosomes and nuclei were prepared, and the proteins were resolved by polyacrylamide gel electrophoresis. The level of incorporation of 32P 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

307

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

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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

308

Induction of apoptosis in human cervical carcinoma HeLa cells by polymethoxylated flavone-rich Citrus grandis Osbeck (Dangyuja) leaf extract.  

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Citrus grandis Osbeck (Dangyuja) has a high content of flavonoids with health-related properties. Although previous data have revealed the anticancer potency of some Citrus species, the underlying molecular mechanisms of this activity by leaf extracts have not been studied in detail. The purpose of this study was to evaluate the cytotoxic effects of citrus leaves on five human cancer cell lines and to determine the possible mechanisms of cell death elicited by the chloroform fraction (CF) of the Dangyuja leaf. The CF of Dangyuja strongly decreased the survival rate of HeLa cells, among the tested cell lines. CF treatment induced the down-regulation of anti-apoptotic Bcl-2 expression, resulting in the proteolytic activation of caspases and the degradation of poly (ADP-ribose) polymerase (PARP) protein. Arrested cell growth and induction of apoptosis were confirmed by flow cytometry and DNA fragmentation analysis, respectively. The major components of the CF were identified as isosinensetin, sinensetin, tetramethyl-O-isoscutellarein, nobiletin, tangeretin, and 5-hydroxy-6,7,8,3',4'-pentamethoxyflavone by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS). Our results suggest that the CF of Dangyuja leaves is an excellent source of functional polymethoxylated flavones, which may help prevent cervical cancer and may potentially be a useful agent for the treatment of certain malignancies. PMID:20538032

Kim, Hana; Moon, Jeong Yong; Mosaddik, Ashik; Cho, Somi Kim

2010-01-01

309

Targeting of the photocytotoxic compound AlPcS4 to Hela cells by transferrin conjugated PEG-liposomes.  

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Photodynamic therapy has attracted increasing interest over the last few years, whereby the activation of photosensitizers by light causes the production of reactive oxygen species (ROS), such as singlet oxygen, which are cytotoxic. The goal of our study was to enhance the photodynamic activity of the photosensitizer aluminum phthalocyanine tetrasulfonate (AlPcS4) through its specific delivery to tumor cells. Since many tumor cells, among which are HeLa cells, overexpress the transferrin receptor, we synthesized transferrin conjugated PEG-liposomes that contained AlPcS4 that could be internalized by receptor mediated endocytosis. The antiproliferative activity of the targeted liposomes was evaluated and compared to the native AlPcS4 and the non-targeted liposome. These findings were supplemented with data on intracellular concentration of the photo-active compounds. The accumulation together with ROS production after irradiation was visualized by using confocal microscopy to confirm the data found in the antiproliferative and accumulation assay. Tf-Lip-AlPcS4 was 10 times more photocytotoxic (IC(50), 0.63 microM) than free AlPcS4 at a light dose of 45 kJ/m whereas Lip-AlPcS4 displayed no photocytotoxicity at all. The high photocytotoxicity of Tf-Lip-AlPcS4 was shown to be the result of a high intracellular concentration (136.5 microM) in HeLa cells, which could be lowered dramatically by incubating the conjugate with a competing transferrin concentration. The images of intracellular accumulation and ROS production matched the accumulation and photocytotoxicity profile of the different photo-active compounds. The photodynamic activity of the Tf-Lip-AlPcS4 conjugate on HeLa cells is much more potent than free AlPcS4 as a result of selective transferrin receptor mediated uptake. PMID:12209592

Gijsens, Antoon; Derycke, Annelies; Missiaen, Ludwig; De Vos, Dirk; Huwyler, Jörg; Eberle, Alex; de Witte, Peter

2002-09-01

310

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 ?B (binding site for NF?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 ?m by using the PDMS (poly(dimethylsiloxane)) stencil technique. The cellular array was first exposed to 100 ng mL-1 dexamethasone, 10 ng mL-1 forskolin, or 100 ng mL-1 TNF-? (tumor necrosis factor ?) 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?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) conventional CL (chemiluminescence) protocol with at least 5 x 104 cells per well.

311

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

312

Basal cell cancer  

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In this chapter authors investigate the peculiarities of basal cell cancer, the theory of it's descent, quote some statistical data on basal cell cancer sickness and variety clinical forms of basal cell cancer was shown

313

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

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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 ...

314

Lethal response of HeLa cells to x irradiation in the latter part of the generation cycle  

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The age-response for the killing of HeLa S3 cells by x rays during the latter part of the generation cycle has been examined in detail. As synchronous cells move from the G1/S boundary through S phase, the relatively high sensitivity of late G1 cells gradually decreases; minimum sensitivity is reached in mid-S and maintained during the remainder of that phase. The response of cells as they progress from S to the point in G2 at which they are temporarily arrested by radiation (or by inhibitors of protein synthesis) was measured in populations free of both S phase cells and late G2 cells that had passed the arrest point: cells retain their high resistance from early G2 up to the arrest point. The response of G2 cells that have passed the arrest point before being irradiated was examined by exposing randomly growing cultures to x rays and collecting cells periodically thereafter, as they entered mitosis. Survival values very close to those of sensitive mitotic cells were found in the 2 h period after irradiation during which unarrested cells continued to reach mitosis. Values typical of late S/early G2 were found only after cells that had been arrested began arriving at mitosis. Thus, HeLa S3 cells undergo an abrupt increase in sensitivity at or near the arrest point. The sensitivity to a second irradiation of cells arrested in G2 by a conditioning x-ray dose increases rapidly in the early part of the arrest period

315

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)

316

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

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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.

Maria T. V., Romanos; Maria J., Andrada-Serpa; Paulo A. S., Mourão; Yocie, Yoneshigue-Valentin; Mariana S., Pereira; Norma, Santos; Marcia D., Wigg.

317

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

318

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

319

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

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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.

320

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

 
 
 
 
321

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

322

Enhanced killing of Hela cells pre-labeled with 5-bromodeoxyuridine by monochromatic synchrotron radiation at 0.9 A  

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The HeLa cells labeled with 5-bromodeoxyuridine were irradiated with monochromatic synchrotron radiation at 0.90 A or 1.00 A, below or above the wavelength of the K absorption edge (0.92 A) of bromine. Although non-specific sensitization of labeled cells to the radiation was observed irrespective of the wavelengths, the labeled cells were killed at a higher rate by the irradiation at 0.90 A than at 1.00 A. The nonlabeled cells showed no difference in the sensitivity to the radiation at the two wavelengths. The enhanced lethality of labeled cells at 0.90 A may be inferred to be due to the induction of Auger effect in bromine atom of the DNA selectively absorbed the photon at 0.90 A. Implications of this finding for the photon activation therapy was discussed. (author)

323

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 ?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.)

324

Activation of NADPH oxidase subunit NCF4 induces ROS-mediated EMT signaling in HeLa cells.  

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The epithelial-mesenchymal transition (EMT) is a critical biological process characterized by morphological and behavioral changes in cells. The regulatory and signaling mechanisms of both developmental and pathological EMT have been investigated. Reactive oxygen species (ROS) play a role in early EMT, but the exact mechanism by which ROS are involved is unclear. We investigated ROS-mediated EMT in human HeLa cells. Transforming growth factor beta (TGF-?) treatments lead to dramatic NADPH oxidase 2 (NOX2) inductions in HeLa cells; antioxidant treatment prevented TGF-?-driven EMT. Over-expression of the p40phox subunit (NCF4) led to activation of the NOX2 complex and ROS production. We showed that NOX2 and NOX5 mRNA was increased, along with increased expression of several matrix metalloproteinases (MMPs) in response to NCF4 expression. Moreover, these changes were reversible upon ROS scavenging. Down-regulation of E-cadherin and up-regulation of Snail, Slug and vimentin occurred at the transcriptional level. We also showed that new EMT regulator, YB-1 is a downstream target in ROS-induced EMT. Together, these data suggest that ROS switching is necessary for increased EMT but is not required for the morphological changes that accompany EMT. PMID:24378533

Kim, Young Mee; Cho, Moonjae

2014-04-01

325

Evaluation of biological activities of Physalis peruviana ethanol extracts and expression of Bcl-2 genes in HeLa cells  

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english Physalis species are used in folk medicine for phytotherapeutic properties. The extracts of medicinal plants are known to possess cytotoxic and chemopreventative compounds. In this study we investigated antibacterial, antioxidant, DNA damage preventative properties of Physalis peruviana (golden berr [...] y) on leaf and shoot ethanol extracts and their effects on cytotoxicity of HeLa cells and expression of apoptotic pathway genes. Among the tested bacteria for antibacterial activity, maximum inhibition zone was determined in Lactococcus lactis. The phenolic content was found higher in leaf extracts than shoot extracts. The antioxidant activity showed the highest TEAC values of the leaf (2 mg/mL) and the shoot (0.5 mg/mL) extracts as 0.291±0.04 and 0.192±0.015, respectively. In DNA damage prevention assay both leaf and shoot extracts, especially 30 and 20 µg/mL concentrations, exhibited significant protection against DNA damage-induced by hydroxyl radical generated by Fenton reaction. Our results suggest that leaf and shoot extracts possess cytotoxic effect on HeLa cells when applied as 100 µg/mL concentration. Also mRNA expression analysis showed the alteration of antiapoptotic genes, so the results suggest that P. peruviana ethanol extracts induce apoptotic cell death and should be investigated for identification of active compounds and their mechanisms of action.

Özgür, Çakir; Murat, Pekmez; Elif, Çepni; Bilgin, Candar; Kerem, Fidan.

2014-06-01

326

Apoptosis in HeLa cell exposed to different dose, dose-rate of 32P ?-irradiation and the correlation with cell-killing efficacy  

International Nuclear Information System (INIS)

In an attempt to elucidate some aspects of the radiobiological basis of targeted radiotherapy in oncology, the apoptosis occurred have been studied in Hela cell lines after exposing to different doses and dose-rate radiation of 32P and the relationship between apoptosis occurred and the capacity of cell proliferation, which might be of help to the understanding of targeted radiotherapy. Asynchronous Hela cells were exposed to ? radiation from 32P absorbed in filter papers which were put closely under culture dishes of growing monolayer of Hela cell. The radiation response characteristics to different dose, dose-rate and radiation time were evaluated through cell-proliferation assessed by the colony-forming assay, cell cycle perturbation studied by flow cytometry and quantity analysis of apoptosis analyzed by flow cytometry and fluorescence microscopy. Morphological and flow cytometry analysis showed a delayed apoptosis. The programmed cell death approached a plateau between 48-72h post-irradiation. Electron and fluorescence microscopic studies showed the presence of morphologically apoptotic cells. Single dose radiation showed a higher apoptosis ratio than multiple low dose radiation, which did not correlate with clonal-forming assay, suggesting apoptosis ratio at a near time point post-irradiation is not a convincing indicator of radiation efficacy in the current experimental setting

327

Isolation of an arabinogalactan from Endopleura uchi bark decoction and its effect on HeLa cells.  

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Endopleura uchi is a native plant from the Amazon used in popular medicine to treat myomas. A crude polysaccharide (AGb) obtained from E. uchi bark decoction was purified, yielding a type II arabinogalactan (AG) that was characterized by chemical and spectroscopic methods. AG was evaluated for its cytotoxic effects on HeLa cells. AG (5-500 ?g/ml) reduced cell viability at 48 and 72 h (approximately 20%) but not in a dose-dependent manner. Cell proliferation was also reduced by AG, with a 25% inhibition (100 ?g/ml) at 72 h. The results suggest that the cytotoxicity exhibited by AG does not involve pathways related to the cell cycle. PMID:24299850

Bento, João Francisco; Noleto, Guilhermina Rodrigues; de Oliveira Petkowicz, Carmen Lúcia

2014-01-30

328

A haemagglutinating adhesin of group B streptococci isolated from cases of bovine mastitis mediates adherence to HeLa cells.  

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Rabbit erythrocytes were agglutinated by 43.4% of group B streptococci isolated from bovines but by none isolated from humans. Haemagglutination was enhanced by cultivation of the bacteria under microaerophilic conditions. Most of the haemagglutinating strains had protein type antigen X, either alone, or in combination with polysaccharide antigens. Heat and proteolytic treatment of the bacteria destroyed the haemagglutination activity. The haemagglutinin could be solubilized from the bacterial surface by mutanolysin treatment and isolated from culture supernatant fluid by ammonium sulphate precipitation. The isolated haemagglutinin did not cause direct agglutination of erythrocytes. However, binding of the haemagglutinin to rabbit erythrocytes could be visualized by agglutination of haemagglutinin-treated erythrocytes by specific antiserum obtained by absorption. Western blotting showed that the haemagglutinin obtained from erythrocyte lysates contained an antibody-reactive band with a molecular mass of 43 kDa. Haemagglutination-positive strains adhered to HeLa cells in higher numbers than did haemagglutination-negative strains. The HeLa cell adherence of Group B streptococci was inhibited in the presence of isolated haemagglutinin or of specific antiserum against the haemagglutinin. These observations suggest that the haemagglutinating adhesins of bovine group B streptococcal isolates are directly involved in the adherence mechanisms of these organisms. PMID:8245843

Wibawan, I W; Lämmler, C; Seleim, R S; Pasaribu, F H

1993-09-01

329

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 T1-resistant oligonucleotide VPg-pUUAAAACAGp or by fingerprint analysis of the in vitro-synthesized 32P-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

330

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

331

Time course of fluorescence intensity and protein expression in HeLa cells stably transfected with hrGFP.  

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The green fluorescent protein (GFP), obtained from Aequorea victoria, is widely used in transfection experiments to study the dynamics and characteristics of gene expression in a non-invasive manner. However, generation of cell lines displaying long-term expression of the GFP protein has repeatedly been reported unsuccessful. In the present study a different marker protein, humanized GFP (hrGFP) obtained from a sea pansy, Renilla reniformis, was used for transfection, and the time course of protein expression was studied by live cell fluorescence microscopy, PCR technology, and immunoblotting. HeLa cells were transfected with a plasmid, phrGFP-IRES-Neo containing the neomycin cassette and the hrGFP gene together with an internal ribosome entry site (IRES) sequence under the control of a CMV promotor. Cells were selected for three weeks after transfection using G418. During this time, hrGFP fluorescence was highest around day 4 and decreased thereafter eventually disappearing until the end of the selection period. In parallel, the proportion of cells staining positive for propidium iodide (i.e. dead cells) increased steadily. Three weeks after transfection non-fluorescent clones were present throughout. A couple of clones were subcultured for another week and used for further analyses. PCR methodology revealed the insertion of the hrGFP into the HeLa genome and the presence of mRNA species coding for hrGFP following reverse transcription. However, immunoblot analysis failed to show expression of the protein. These observations suggest a down-regulation of hrGFP expression at both transcriptional and post-transcriptional levels similar to results previously obtained with GFP. Thus, use of hrGFP does not appear to offer an advantage over GFP in transfection experiments aiming at permanent expression of the marker protein. PMID:12872990

Kirsch, Petra; Hafner, Mathias; Zentgraf, Hanswalter; Schilling, Lothar

2003-06-30

332

Design and Synthesis of New Chacones Substituted with Azide/Triazole Groups and Analysis of Their Cytotoxicity Towards HeLa Cells  

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Full Text Available A series of new chalcones substituted with azide/triazole groups were designed and synthesized, and their cytotoxic activity was evaluated in vitro against the HeLa cell line. O-Alkylation, Claisen-Schmidt condensation and Cu(I-catalyzed cycloaddition of azides with terminal alkynes were applied in key steps. Fifteen compounds were tested against HeLa cells. Compound 8c was the most active molecule, with an IC50 value of 13.03 µM, similar to the value of cisplatin (7.37 µM.

José A. F. P. Villar

2012-08-01

333

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

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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.)

334

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

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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

335

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

336

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

337

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

338

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 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

Aurea S., Cruz; Cristina A., Figueiredo; Clélia H. O., Martinez; Luís F. de, Salles Gomes.

339

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

Aurea S., Cruz; Cristina A., Figueiredo; Clélia H. O., Martinez; Luís F. de, Salles Gomes.

1992-04-01

340

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

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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

 
 
 
 
341

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

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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.

Narasimha Rao, M.V.

1987-03-01

342

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

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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

343

Involvement of aldolase A in X-ray resistance of human HeLa and UVr-1 cells  

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To find novel proteins involved in radio-resistance of human cells, we searched for nuclear proteins, whose expression levels alter after X-ray irradiation in HeLa cells, using agarose fluorescent two-dimensional differential gel electrophoresis following mass spectrometry. We identified 6 proteins, whose levels were increased in nuclei 24 h after irradiation at 5 Gy, including aldolase A. Nuclear aldolase A levels increased twofold after the irradiation, however, total aldolase A levels did not change. When the expression of aldolase A was suppressed by its specific siRNA, sensitization of the suppressed cells to X-ray-induced cell death was observed. In addition, UVr-1 cells with higher aldolase A expression exhibited lower sensitivity to X-ray-induced cell death than the parental RSa cells with lower aldolase A expression. These results suggest that aldolase A may play a role in the radio-response of human cells, probably in nuclei, in addition to its glycolytic role in the cytosol

344

Depletion of mitochondrial DNA by down-regulation of deoxyguanosine kinase expression in non-proliferating HeLa cells  

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Purine deoxyribonucleotides required for mitochondrial DNA replication are either imported from the cytosol or derived from phosphorylation of deoxyadenosine or deoxyguanosine catalyzed by mitochondrial deoxyguanosine kinase (DGUOK). DGUOK deficiency has been linked to mitochondrial DNA depletion syndromes suggesting an important role for this enzyme in dNTP supply. We have generated HeLa cell lines with 20-30% decreased levels of DGUOK mRNA by the expression of small interfering RNAs directed towards the DGUOK mRNA. The cells with decreased expression of the enzyme showed similar levels of mtDNA as control cells when grown exponentially in culture. However, mtDNA levels rapidly decreased in the cells when cell cycle arrest was induced by serum starvation. DNA incorporation of 9-?-D-arabino-furanosylguanine (araG) was lower in the cells with decreased deoxyguanosine kinase expression, but the total rate of araG phosphorylation was increased in the cells. The increase in araG phosphorylation was shown to be due to increased expression of deoxycytidine kinase. In summary, our findings show that DGUOK is required for mitochondrial DNA replication in resting cells and that small changes in expression of this enzyme may cause mitochondrial DNA depletion. Our data also suggest that alterations in the expression level of DGUOK may induce compensatory changes in the expression of other nucleoside kinases

345

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.

346

Radioadaptive response to the medium-mediated bystander induction of DNA strand breaks in HeLa cells  

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Full text: Numerous investigators have reported two cellular responses of importance at low doses that have a potential impact on the risk estimation of ionizing radiation. The radioadaptive response confers resistance to a subsequent dose by a low priming dose, while the bystander effect exaggerates the effect of small doses. The present study was conducted to examine the interaction of the radioadaptive response with the bystander effect in HeLa cells. The culture was irradiated with 0.5 to 8 Gy of 140 kVp X-rays and one hour later, the medium was taken, passed through a filter and transferred to the parallel culture of non-irradiated HeLa cells as non-targeted cells. After incubation for 30 min, the induced DNA damage was analyzed by the single cell gel-electrophoresis assay under alkaline or neutral conditions. The treatments resulted in a dose-dependent increase in tail moment under either conditions, indicating the induction of DNA single- and double-strand breaks. The clonogenic survival of non-irradiated cells was also reduced after they were cultured in the medium that was taken from irradiated cultures. Any change was not observed when the medium alone was irradiated. These results give the disputed evidence that certain genotoxic factor(s) released from irradiated cells into the culture medium can induce DNA strand breaks leading to cell death. It is also suggested that physical contact between irradiated and non-irradiated cells may not be required for theradiated cells may not be required for the bystander effect. In adapted cells that were pre-exposed to 5 cGy of X-rays and cultured for 4 h beforehand, the yield of DNA strand breaks induced by X-rayed medium was reduced by about 50 %. The results, in conjunction with our early finding (Ikushima et al., 1996) suggest that the radioadaptive response resulting from such a low dose may diminish the bystander effect through an enhanced DNA repair function

347

Picosecond pulsed electric fields induce apoptosis in HeLa cells via the endoplasmic reticulum stress and caspase-dependent signaling pathways.  

Science.gov (United States)

The non-invasive treatment of tumors with preserved fertility holds great promise. The application of pulsed electric field (PEF) is a new biomedical engineering technique for tumor therapy. Picosecond pulsed electric fields (psPEF) can be transferred to target deep tissue non-invasively and precisely; however, research of the biological effects of psPEF on cells is limited. Electric theory predicts that when the pulse duration decreases to nanoseconds and picoseconds, it will mainly affect organelles and lead to intracellular electromanipulations. Previous studies have shown that psPEF targets the mitochondria and induces apoptosis through a mitochondrial-mediated pathway in HeLa cells. The endoplasmic reticulum is also involved in the intrinsic pathways of apoptosis. In the present study, HeLa cells were exposed to psPEF to investigate the underlying mechanisms of apoptosis. MTT assay demonstrated that psPEF displayed strong growth inhibitory effects on HeLa cells. Treatment with psPEF led to marked cell apoptosis and cell cycle arrest at the G2/M phase. In addition, psPEF affected the phosphorylation levels of endoplasmic reticulum sensors and upregulated the expression of glucose-regulated protein 78 (GRP78), glucose-regulated protein 94 (GRP94) and CCAAT enhancer-binding protein (C/EBP) homologous protein (CHOP). These changes were accompanied by the elevation of intracellular Ca2+ concentrations. Furthermore, the activation of caspase-12, -9 and -3, led to the release of cytochrome c, as well as the upregulation of Bax and the downregulation of Bcl-2, as observed in the HeLa cells. Taken together, these data suggest that psPEF is an efficient apoptosis-inducing agent for HeLa cells, which exerts its effects, at least partially, via the endoplasmic reticulum stress and caspase-dependent signaling pathways. PMID:23338860

Chen, Wen-Juan; Xiong, Zheng-Ai; Zhang, Min; Yao, Chen-Guo; Zhao, Zhong-Yong; Hua, Yuan-Yuan; Zhou, Wei

2013-03-01

348

Alphacoronavirus transmissible gastroenteritis virus nsp1 protein suppresses protein translation in mammalian cells and in cell-free HeLa cell extracts but not in rabbit reticulocyte lysate.  

Science.gov (United States)

The nsp1 protein of transmissible gastroenteritis virus (TGEV), an alphacoronavirus, efficiently suppressed protein synthesis in mammalian cells. Unlike the nsp1 protein of severe acute respiratory syndrome coronavirus, a betacoronavirus, the TGEV nsp1 protein was unable to bind 40S ribosomal subunits or promote host mRNA degradation. TGEV nsp1 also suppressed protein translation in cell-free HeLa cell extract; however, it did not affect translation in rabbit reticulocyte lysate (RRL). Our data suggested that HeLa cell extracts and cultured host cells, but not RRL, contain a host factor(s) that is essential for TGEV nsp1-induced translational suppression. PMID:21047955

Huang, Cheng; Lokugamage, Kumari G; Rozovics, Janet M; Narayanan, Krishna; Semler, Bert L; Makino, Shinji

2011-01-01

349

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

350

Nucleotide sequence of a cDNA for an apurinic/apyrimidinic endonuclease from HeLa cells  

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A cDNA, HAP1h, encoding a human apurinic/apyrimidinic (AP) endonuclease from HeLa cells has been cloned and sequenced. The predicted HAP1h protein consists of 318 amino acids and shows 97% similarity and 93% identity to a bovine AP endonuclease (BAP1). While this work was in progress, the HAP1 cDNA sequence, which also encodes a protein homologous to the BAP1 protein, was reported. The HAp1 cDAN was isolated from a human melanoma cDNA library. A comparison of the two cDNA sequences (HAP1h and HAP1) revealed two differences in the coding region (CR1 and CR2) and one in the 5{prime}-untranslated region (5'UTR).

Cheng, Xinbo; Bunville, J.; Patterson, T.A. (DuPont Merck Pharmaceutical Co., Wilmington, DE (United States))

1992-01-25

351

The use of 1H spin echo NMR and HPLC to confirm doxorubicin induced depletion of glutathione in the intact HeLa cell.  

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The effects of doxorubicin on the cellular biochemistry of the HeLa cell using 1H spin echo nuclear magnetic resonance spectroscopy (NMR) of the intact and viable cell in conjunction with dual wavelength HPLC of cell lysates is reported. Directly dose-related changes were observed in lactate and reduced glutathione concentration. Doxorubicin induces a time-dependent depletion of the cytosolic pool of glutathione and a change in the glycolytic pattern of the cell. The glutathione depletion cou...

Al-kabban, M.; Watson, I. D.; Stewart, M. J.; Reglinski, J.; Smith, W. E.; Suckling, C. J.

1988-01-01

352

Squamous cell cancer (image)  

Science.gov (United States)

... a malignant tumor, and is more aggressive than basal cell cancer, but still may be relatively slow-growing. It is more likely than basal cell cancer to spread (metastasize) to other locations, including internal ...

353

Effect of Ureaplasma parvum co-incubation on Chlamydia trachomatis maturation in human epithelial HeLa cells treated with interferon-?.  

Science.gov (United States)

Chlamydia trachomatis is an obligate intracellular bacterium that causes a sexually transmitted disease. Ureaplasma parvum is commensal in the human genital tract, with a minimal contribution to urogenital infection. We have recently found that U. parvum has a significant effect on the presence of C. trachomatis in the genital tract of healthy women. We therefore assessed the effect of U. parvum co-incubation on C. trachomatis maturation from reticulate bodies (RBs) to elementary bodies (EBs) in HeLa cells in the absence or presence of interferon (IFN)-?, which is a critical host defense factor. IFN-? stimulation of viable U. parvum significantly prompted chlamydial growth with an increase in infectious particles, EBs, in HeLa cells. IFN-? treatment of killed U. parvum had a similar effect on C. trachomatis maturation in HeLa cells. There was no change in expression of indoleamine 2,3-dioxygenase (IDO) in cultures of viable or killed U. parvum. We concluded that U. parvum co-incubation by IFN-? helped C. trachomatis to mature from RBs to EBs in HeLa cells, independent of IDO expression. This suggests a novel survival strategy of C. trachomatis against IFN-? exposure, prompting secondary infection of the genital mucosa, with possible clinical implications. PMID:24855914

Yamazaki, Tomohiro; Matsuo, Junji; Nakamura, Shinji; Oguri, Satoshi; Yamaguchi, Hiroyuki

2014-08-01

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Changes in the relative proportion of transformation-sensitive polypeptides in giant HeLa cells produced by irradiation with lethal doses of x-rays  

International Nuclear Information System (INIS)

Irradiation of HeLa cells with 1,100 rads (1 rad = 0.01 J/kg = 0.01 Gy) of x-rays yielded a pure population of giant cells 5-7 days after irradiation. These cells do not divide but go through an intermittent DNA synthetic phase. The population of giant cells in S phase (8%) is considerably lower than that of control asynchronous HeLa cells (30%), but 80% of the giant cells go through S phase as determined by 48-hr labeling with [3H]thymidine. Previous studies with high-resolution two-dimensional gel electrophoreis identified 58 [35S]methionine-labeled polypeptides common to human epithelia amnion cells and lung fibroblats, whose rate of synthesis is sensitive to neoplastic transformation. These polypeptides also have been identified in HeLa cells and other transformed human cells such as Detroit 98, Chang liver, Fl-amnion, and WISH-amnion. After irradiation of HeLa cells and giant cell formation, the relative proportion of most of the transformation-sensitive polypeptides (43 of 47) reverted to levels similar to those observed in nontumorigenic cells. This suggests that their relative proportions are dependent on the growth properties of the cells. In particular, the relative proportions of three polypeptides (designated 12g and 60d1 in isoelectric focusing and 27b in nonequilibrium pH gradient electrophoresis) were not affected, indicating that their reduced amounts in transformed cells could reflect a fundamental change that develops during transformation

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

International Nuclear Information System (INIS)

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 th