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

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Amygdalin, a naturally occurring substance, has been suggested to be efficacious as an anticancer substance. The effect of amygdalin on cervical cancer cells has never been studied. In this study, we found that the viability of human cervical cancer HeLa cell line was significantly inhibited by amygdalin. 4,6-Diamino-2-phenyl indole (DAPI) staining showed that amygdalin-treated HeLa cells developed typical apoptotic changes. The development of apoptosis in the amygdalin-treated HeLa cells were confirmed by double staining of amygdalin-treated HeLa cells with annexin V-FITC and propidium iodide (PI) along with increase in caspase-3 activity in these cells. Further studies indicated that antiapoptotic protein Bcl-2 was downregulated whereas proapoptotic Bax protein was upregulated in the amygdalin-treated HeLa cells implying involvement of the intrinsic pathway of apoptosis. In vivo, amygdalin administration inhibited the growth of HeLa cell xenografts through a mechanism of apoptosis. The results in the present study suggest that amygdalin may offer a new therapeutic option for patients with cervical cancer. PMID:23137229

Chen, Yu; Ma, Jinshu; Wang, Fang; Hu, Jie; Cui, Ai; Wei, Chengguo; Yang, Qing; Li, Fan

2013-02-01

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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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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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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% tn 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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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.

Liu, Junye [Department of Radiation Medicine, Fourth Military Medical University, Xi' an (China); Department of Biochemistry and Molecular Biology, State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi' an (China); Zhang, Jing [Department of Biochemistry and Molecular Biology, State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi' an (China); Wang, Xiaowu [Department of Radiation Medicine, Fourth Military Medical University, Xi' an (China); Li, Yan [Department of Biochemistry and Molecular Biology, State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi' an (China); Chen, Yongbin; Li, Kangchu [Department of Radiation Medicine, Fourth Military Medical University, Xi' an (China); Zhang, Jian [Department of Biochemistry and Molecular Biology, State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi' an (China); Yao, Libo, E-mail: bioyao@fmmu.edu.cn [Department of Biochemistry and Molecular Biology, State Key Laboratory of Cancer Biology, Fourth Military Medical University, Xi' an (China); Guo, Guozhen, E-mail: guozhengg@hotmail.com [Department of Radiation Medicine, Fourth Military Medical University, Xi' an (China)

2010-07-15

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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 ( 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)

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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 and Mechanism of Baicalin on Apoptosis of Cervical Cancer HeLa Cells I n -v itro.  

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The objective of this study was to observe the apoptosis-inducing effect and mechanism of baicalin on human cervical cancer HeLa cells. The inhibitory effect of baicalin on the growth of HeLa cells was measured by MTT assay, and cell proliferation and migration was analyzed by cell scratch assay. Morphological changes of apoptotic cells were viewed by the light microscope and electron microscope, and cell growth arrest was confirmed by flow cytometry. Moreover, Western blot was used for investigating the expression of apoptosis related proteins; spectrophotometry was used to examine Caspase-3 activation. Our results showed that baicalin could inhibit the proliferation of HeLa Cells via induction of apoptosis in a time and dose-dependent manner (P<0.01). Apoptotic signaling induced by baicalin was characterized by up-regulating Bax, Fas, FasL and Caspase-8 protein expression, and down-regulating of Bcl-2 protein expression. These results indicated that baicalin-induced apoptosis involved activation Caspase-3 in HeLa cells through the intracellular mitochondrial pathway and the surface death receptor pathway. PMID:25561931

Peng, Yong; Fu, Zhan-Zhao; Guo, Cong-Shan; Zhang, Yan-Xia; Di, Ya; Jiang, Bin; Li, Qing-Wang

2015-01-01

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Effects and Mechanism of Baicalin on Apoptosis of Cervical Cancer HeLa Cells In-vitro  

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The objective of this study was to observe the apoptosis-inducing effect and mechanism of baicalin on human cervical cancer HeLa cells. The inhibitory effect of baicalin on the growth of HeLa cells was measured by MTT assay, and cell proliferation and migration was analyzed by cell scratch assay. Morphological changes of apoptotic cells were viewed by the light microscope and electron microscope, and cell growth arrest was confirmed by flow cytometry. Moreover, Western blot was used for investigating the expression of apoptosis related proteins; spectrophotometry was used to examine Caspase-3 activation. Our results showed that baicalin could inhibit the proliferation of HeLa Cells via induction of apoptosis in a time and dose-dependent manner (P<0.01). Apoptotic signaling induced by baicalin was characterized by up-regulating Bax, Fas, FasL and Caspase-8 protein expression, and down-regulating of Bcl-2 protein expression. These results indicated that baicalin-induced apoptosis involved activation Caspase-3 in HeLa cells through the intracellular mitochondrial pathway and the surface death receptor pathway.

Peng, Yong; Fu, Zhan-zhao; Guo, Cong-Shan; Zhang, Yan-Xia; Di, Ya; Jiang, Bin; Li, Qing-Wang

2015-01-01

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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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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)

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

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

Abbas Zare Mirakabadi

2012-10-01

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

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

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

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Objective: To detect the effects of ionizing radiation combined with autophagy inhibitors and inducers on the proliferation of human cervical cancer cell line. Methods: MTT and flowcytometry (FCM) were used to detect the surviving and proliferation of human cervical cancer cells,and analysis of the relationship of dose-effect and time-effect was made. Results: With the increase of irradiation doses (2, 4, 6, 8 and 10 Gy) and the elongation of irradiation time (24, 48 and 72 h), the inhibiting effect of ionizing radiation on the proliferation of human cervical cancer cells increased (P< 0.05 or P< 0.01). The inhibiting effect of 6 Gy combined with autophagy inducer rapamycin on the proliferation of Hela cells weakened (P< 0.05). The inhibiting effects of 6 Gy combined with autophagy inhibitor 3-MA on the cell proliferation were higher than those in 6 Gy group (P< 0.05). Conclusion: Ionizing radiation combined with autophagy inducers can inhibit apoptosis in Hela cells, while the ionizing radiation combined with autophagy inhibitors can promote their apoptosis. (authors)

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

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

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

2014-05-01

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

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

Brito, Daniela A.; Rieder, Conly L.

2009-01-01

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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)

20

Effects of a nutrient mixture on immunohistochemical localization of cancer markers in human cervical cancer HeLa cell tumor xenografts in female nude mice  

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Although fully treatable in the early stages, once cervical cancer has metastasized, patient outcome is poor. The main objective of this study was to examine the effect of dietary supplementation with a nutrient mixture (NM) containing lysine, ascorbic acid, proline, green tea extract and other micronutrients on HeLa cell xenografts in nude female mice. Tumor growth was measured and immunohistochemical staining was evaluated for the following cancer markers: Ki67 (proliferation); matrix metalloproteinase (MMP)-2 and -9 (invasion/metastasis); vascular endothelial growth factor (VEGF) (angiogenesis); terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) and B-cell lymphoma 2 (Bcl-2) (apoptosis); cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS) (inflammation); and glutathione S-transferase ? (GST?) (a general cancer marker). Following housing for a week, 5/6-week-old female athymic nude mice (n=12) were inoculated subcutaneously with 3×106 HeLa cells in 0.2 ml phosphate-buffered saline and 0.1 ml Matrigel™ and randomly divided into two groups; control group mice were fed regular mouse chow and NM group mice the regular diet supplemented with 0.5% NM (w/w). After four weeks, the mice were sacrificed and their tumors were excised and processed for histology. The NM strongly inhibited the growth of HeLa xenografts in nude mice. The mean tumor weight was reduced to 59% (P=0.001) in the mice fed the NM compared with the tumor weight in the controlled diet mice. Ki67, MMP-2 and -9, VEGF, TUNEL, Bcl-2, COX-2, iNOS and GST? all showed a lower intensity and frequency of staining in the NM group compared with that in the control group. In conclusion, NM supplementation strongly inhibited tumor growth and cancer markers in female nude mice injected with HeLa xenografts.

ROOMI, M.W.; KALINOVSKY, T.; CHA, J.; ROOMI, N.W.; NIEDZWIECKI, A.; RATH, M.

2015-01-01

21

Physico-chemical characteristics of ZnO nanoparticles-based discs and toxic effect on human cervical cancer HeLa cells  

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In this study, we investigated physico-chemical properties of zinc oxide nanoparticles (ZnO NPs)-based discs and their toxicity on human cervical cancer HeLa cell lines. ZnO NPs (80 nm) were produced by the conventional ceramic processing method. FESEM analysis indicated dominant structure of nanorods with dimensions 100-500 nm in length, and 20-100 nm in diameter. The high content of ZnO nanorods in the discs probably played significant role in toxicity towards HeLa cells. Structural defects (oxygen vacancies and zinc/oxygen interstitials) were revealed by PL spectra peaks at 370-376 nm and 519-533 nm for the ZnO discs. The structural, optical and electrical properties of prepared sample have influenced the toxicological effects of ZnO discs towards HeLa cell lines via the generation of reactive oxygen species (ROS), internalization, membrane damage, and eventually cell death. The larger surface to volume area of the ZnO nanorods, combined with defects, stimulated enhanced toxicity via ROS generation hydrogen peroxide, hydroxyl radicals, and superoxide anion. The preliminary results confirmed the ZnO-disc toxicity on HeLa cells was significantly associated with the unique physicochemical properties of ZnO NPs and to our knowledge, this is the first cellular study for treatment of HeLa cells with ZnO discs made from 80 nm ZnO particles.

Sirelkhatim, Amna; Mahmud, Shahrom; Seeni, Azman; Kaus, Noor Haida Mohd.; Sendi, Rabab

2014-10-01

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DYTOGENETIC ANALYSIS OF HELA AND CHANG CELLS  

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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 ...

Lzadian, N.; Sussman, H.

1982-01-01

23

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

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

Smith Charlotte M

2013-01-01

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

International Nuclear Information System (INIS)

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.

25

Anticancer Effects of Brominated Indole Alkaloid Eudistomin H from Marine Ascidian Eudistoma viride Against Cervical Cancer Cells (HeLa).  

Science.gov (United States)

Marine invertebrates called ascidians are prolific producers of bioactive substances. The ascidian Eudistoma viride, distributed along the Southeast coast of India, was investigated for its in vitro cytotoxic activity against human cervical carcinoma (HeLa) cells by the MTT assay. The crude methanolic extract of E. viride, with an IC50 of 53 ?g/ml, was dose-dependently cytotoxic. It was more potent at 100 ?g/ml than cyclohexamide (1 ?g/ml), reducing cell viability to 9.2%. Among nine fractions separated by chromatography, ECF-8 exhibited prominent cytoxic activity at 10 ?g/ml. The HPLC fraction EHF-21 of ECF-8 was remarkably dose- and time-dependently cytotoxic, with 39.8% viable cells at 1 ?g/ml compared to 51% in cyclohexamide-treated cells at the same concentration; the IC50 was 0.49 ?g/ml. Hoechst staining of HeLa cells treated with EHF-21 at 0.5 ?g/ml revealed apoptotic events such an cell shrinkage, membrane blebbing, chromatin condensation and formation of apoptotic bodies. Cell size and granularity study showed changes in light scatter, indicating the characteristic feature of cells dying by apoptosis. The cell-cycle analysis of HeLa cells treated with fraction EHF-21 at 1 ?g/ml showed the marked arrest of cells in G0/G1, S and G2/M phases and an increase in the sub G0/G1 population indicated an increase in the apoptotic cell population. The statistical analysis of the sub-G1 region showed a dose-dependent induction of apoptosis. DNA fragmentation was also observed in HeLa cells treated with EHF-21. The active EHF-21 fraction, a brominated indole alkaloid Eudistomin H, led to apoptotic death of HeLa cells. PMID:25550562

Rajesh, Rajaian Pushpabai; Annappan, Murugan

2015-01-01

26

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

Science.gov (United States)

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

27

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

Science.gov (United States)

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

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

2010-06-01

28

Lanthanum citrate induces anoikis of Hela cells.  

Science.gov (United States)

Some reports show that lanthanum, a rare earth element, induces apoptosis in certain cancer cells. In the present paper, we report the first observation of anoikis induced by lanthanum citrate (LaCit) in Hela cells at a concentration of 0.001-0.1 mmol/L after 48h-treatment. Before cell treatment, Hela cells were subjected to anoikis-resistant selection to remove anoikis-sensitive cells and ensure specificity of LaCit-induced anoikis. Anoikis was determined by Annexin/PI, AO/EB staining, cleavage of PARP and soft-agar colony forming assay. Further, findings of decreased mitochondrial membrane potential, the cleavage of caspase-9 and a dose-dependent increase expression of Bax were detected, suggesting that the intrinsic caspase pathway was involved in the anoikis induced by LaCit. In addition, activation of caspase-8 occurred later than that of caspase-9. LaCit also caused reorganization of actin cytoskeleton, and was accompanied by an increase in co-localization of F-actin with mitochondria, implying that both actin cytoskeleton and mitochondria may play important roles in LaCit -induced anoikis. PMID:19679391

Su, Xiange; Zheng, Xiaona; Ni, Jiazuan

2009-11-28

29

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

Directory of Open Access Journals (Sweden)

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

K Pooshang Bagheri

2013-06-01

30

DYTOGENETIC ANALYSIS OF HELA AND CHANG CELLS  

Directory of Open Access Journals (Sweden)

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

31

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

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Cervical cancer is one of the most common cancers among women in the world. 6-Shogaol is a natural compound isolated from the rhizome of ginger (Zingiber officinale). In this paper, we demonstrated that 6-shogaol induced apoptosis and G2/M phase arrest in human cervical cancer HeLa cells. Endoplasmic reticulum stress and mitochondrial pathway were involved in 6-shogaol-mediated apoptosis. Proteomic analysis based on label-free strategy by liquid chromatography chip quadrupole time-of-flight mass spectrometry was subsequently proposed to identify, in a non-target-biased manner, the molecular changes in cellular proteins in response to 6-shogaol treatment. A total of 287 proteins were differentially expressed in response to 24 h treatment with 15 ?M 6-shogaol in HeLa cells. Significantly changed proteins were subjected to functional pathway analysis by multiple analyzing software. Ingenuity pathway analysis (IPA) suggested that 14-3-3 signaling is a predominant canonical pathway involved in networks which may be significantly associated with the process of apoptosis and G2/M cell cycle arrest induced by 6-shogaol. In conclusion, this work developed an unbiased protein analysis strategy by shotgun proteomics and bioinformatics analysis. Data observed provide a comprehensive analysis of the 6-shogaol-treated HeLa cell proteome and reveal protein alterations that are associated with its anticancer mechanism. PMID:23243437

Liu, Qun; Peng, Yong-Bo; Qi, Lian-Wen; Cheng, Xiao-Lan; Xu, Xiao-Jun; Liu, Le-Le; Liu, E-Hu; Li, Ping

2012-01-01

32

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

OpenAIRE

Cervical cancer is one of the most common cancers among women in the world. 6-Shogaol is a natural compound isolated from the rhizome of ginger (Zingiber officinale). In this paper, we demonstrated that 6-shogaol induced apoptosis and G2/M phase arrest in human cervical cancer HeLa cells. Endoplasmic reticulum stress and mitochondrial pathway were involved in 6-shogaol-mediated apoptosis. Proteomic analysis based on label-free strategy by liquid chromatography chip quadrupole time-of-flight m...

Qun Liu; Yong-Bo Peng; Lian-Wen Qi; Xiao-Lan Cheng; Xiao-Jun Xu; Le-Le Liu; E-Hu Liu; Ping Li

2012-01-01

33

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

Science.gov (United States)

[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

34

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

International Nuclear Information System (INIS)

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

35

The potentized homeopathic drug, Lycopodium clavatum (5C and 15C) has anti-cancer effect on hela cells in vitro.  

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

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

2013-08-01

36

Latex of Euphorbia antiquorum-induced S-phase arrest via active ATM kinase and MAPK pathways in human cervical cancer HeLa cells.  

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Latex of Euphorbia antiquorum (EA) has demonstrated great chemotherapeutic potential for cancer. However, the mechanisms of anti-proliferation of EA on cancer cell remain to be further investigated. The purpose of this study was to explore the influence of EA in human cervical cancer cells. Here, the cell cycle distribution by flow cytometry was examined and the protein expression by the western blotting methods was analyzed. From the cytometric results it was shown that EA-induced S-phase arrest in a concentration manner both in human cervical cancer HeLa and CaSki cells. According the western blot results it was illustrated that EA could downregulate early cyclin E1-Cdk2; and cyclin A-Cdc2 provides a significant additional quantity of S-phase promotion, that in turn promoted the expression of p21(waf1/cip1) and p27(kip1) which were the inhibitors in the complex of cyclin A and Cdc2 that led to cell cycle arrest. Moreover, EA promoted the activation of ataxia telangiectasia mutated (ATM) and check-point kinase-2 (Chk2); however, it negatively regulated the expression of Topoisomerases I and II, Cdc25A, and Cdc25C signaling. Caffeine, an ATM/ATR inhibitor significantly reversed EA downregulation in the levels of Cdc25A. Furthermore, JNK inhibitor SP600125 and p38 MAPK inhibitor SB203580 both could reverse the EA upregulation of the protein of Chk2 level, significantly. This study, therefore, revealed that EA could downregulate topoisomerase, and activate ATM kinase, which then induce parallel Chk 1/2 and MAPK signaling pathways to promote the degradation of Cdc25A to induced S-phase arrest in human cervical cancer HeLa cells. © 2014 Wiley Periodicals, Inc. Environ Toxicol, 2014. PMID:24706497

Hsieh, Wen-Tsong; Lin, Hui-Yi; Chen, Jou-Hsuan; Lin, Wen-Chung; Kuo, Yueh-Hsiung; Wood, W Gibson; Lu, Hsu-Feng; Chung, Jing-Gung

2014-04-01

37

Oxidative stress induced by copper and iron complexes with 8-hydroxyquinoline derivatives causes paraptotic death of HeLa cancer cells.  

Science.gov (United States)

Here, we report the antiproliferative/cytotoxic properties of 8-hydroxyquinoline (8-HQ) derivatives on HeLa cells in the presence of transition metal ions (Cu(2+), Fe(3+), Co(2+), Ni(2+)). Two series of ligands were tested, the arylvinylquinolinic L1-L8 and the arylethylenequinolinic L9-L16, which can all interact with metal ions by virtue of the N,O donor set of 8-HQ; however, only L9-L16 are flexible enough to bind the metal in a multidentate fashion, thus exploiting the additional donor functions. L1-L16 were tested for their cytotoxicity on HeLa cancer cells, both in the absence and in the presence of copper. Among them, the symmetric L14 exhibits the highest differential activity between the ligand alone (IC50 = 23.7 ?M) and its copper complex (IC50 = 1.8 ?M). This latter, besides causing a significant reduction of cell viability, is associated with a considerable accumulation of the metal inside the cells. Metal accumulation is also observed when the cells are incubated with L14 complexed with other late transition metal ions (Fe(3+), Co(2+), Ni(2+)), although the biological response of HeLa cells is different. In fact, while Ni/L14 and Co/L14 exert a cytostatic effect, both Cu/L14 and Fe/L14 trigger a caspase-independent paraptotic process, which results from the induction of a severe oxidative stress and the unfolded protein response. PMID:24592930

Barilli, Amelia; Atzeri, Corrado; Bassanetti, Irene; Ingoglia, Filippo; Dall'Asta, Valeria; Bussolati, Ovidio; Maffini, Monica; Mucchino, Claudio; Marchiò, Luciano

2014-04-01

38

Irradiation And Papillomavirus E2 Proteins On Hela Cells  

International Nuclear Information System (INIS)

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

39

Identification of human patatin-like phospholipase domain-containing protein 1 and a mutant in human cervical cancer HeLa cells.  

Science.gov (United States)

Recently members of mammalian patatin-like phospholipase domain containing (PNPLA) protein family have attracted attention for their critical roles in diverse aspects of lipid metabolism and signal pathway. Until now little has been known about the characteristics of PNPLA1. Here, the full length coding cDNA sequence of human PNPLA1 (hPNPLA1) was cloned for the first time, which encoded a polypeptide with 532 amino acids containing the whole patatin domain. Tissue expression profiles analysis showed that low mRNA levels of hPNPLA1 existed in various tissues, except high expression in the digestive system, bone marrow and spleen. Subcellular distribution of hPNPLA1 tagged with green fluorescence protein mainly localized to lipid droplets. Furthermore, a nonsense mutation of PNPLA1 in human cervical cancer HeLa cells was identified. The hPNPLA1 mutant encoded a protein of 412 amino acids without the C-terminal domain and did not colocalize to lipid droplets, which suggested that the C-terminal region of hPNPLA1 affected lipid droplet binding. These results identified hPNPLA1 and a mutant in HeLa cells, and provided insights into the structure and function of PNPLA1. PMID:24057234

Chang, Ping-An; Sun, Ying-Jian; Huang, Fei-Fei; Qin, Wen-Zhen; Chen, Yu-Ying; Zeng, Xin; Wu, Yi-Jun

2013-10-01

40

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  

OpenAIRE

Jatropha meal produced from the kernel of Jatropha curcas Linn. grown in Malaysia contains phorbol esters (PEs). The potential benefits of PEs present in the meal as anticancer agent are still not well understood. Hence, this study was conducted to evaluate the cytotoxic effects and mode of actions of PEs isolated from Jatropha meal against breast (MCF-7) and cervical (HeLa) cancer cell lines. Isolated PEs inhibited cells proliferation in a dose-dependent manner of both MCF-7 and HeLa cell li...

Syahida Ahmad; Norhani Abdullah; Ehsan Oskoueian

2012-01-01

41

Photodynamic Effects of Pterin on HeLa Cells  

DEFF Research Database (Denmark)

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

Denofrio, M. Paula; Lorente, Carolina

2011-01-01

42

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

Science.gov (United States)

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

Rajasekharreddy, Pala; Rani, Pathipati Usha

2014-06-01

43

Effect of Quercetin on radio-sensitivity of HeLa cells  

International Nuclear Information System (INIS)

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)

44

Thymosin Beta-4, Actin-Sequestering Protein Regulates Vascular Endothelial Growth Factor Expression via Hypoxia-Inducible Nitric Oxide Production in HeLa Cervical Cancer Cells  

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Vascular endothelial growth factor (VEGF) is an important regulator of neovascularization. Hypoxia inducible nitric oxide (NO) enhanced the expression of VEGF and thymosin beta-4 (T?4), actin sequestering protein. Here, we investigated whether NO-mediated VEGF expression could be regulated by T?4 expression in HeLa cervical cancer cells. Hypoxia inducible NO production and VEGF expression were reduced by small interference (si) RNA of T?4. Hypoxia response element (HRE)-luciferase activity and VEGF expression were increased by the treatment with N-(?-D-Glucopyranosyl)-N2-acetyl-S-nitroso-D, L-penicillaminamide (SNAP-1), to generate NO, which was inhibited by the inhibition of T?4 expression with T?4-siRNA. In hypoxic condition, HRE-luciferase activity and VEGF expression were inhibited by the treatment with NG-monomethyl-L-arginine (L-NMMA), an inhibitor to nitric oxide synthase (NOS), which is accompanied with a decrease in T?4 expression. VEGF expression inhibited by L-NMMA treatment was restored by the transfection with pCMV-T?4 plasmids for T?4 overexpression. Taken together, these results suggest that T?4 could be a regulator for the expression of VEGF via the maintenance of NOS activity.

Ryu, Yun-Kyoung; Lee, Jae-Wook; Moon, Eun-Yi

2015-01-01

45

Thymoquinone-Loaded Nanostructured Lipid Carrier Exhibited Cytotoxicity towards Breast Cancer Cell Lines (MDA-MB-231 and MCF-7) and Cervical Cancer Cell Lines (HeLa and SiHa)  

Science.gov (United States)

Thymoquinone (TQ) has been shown to exhibit antitumor properties. Thymoquinone-loaded nanostructured lipid carrier (TQ-NLC) was developed to improve the bioavailability and cytotoxicity of TQ. This study was conducted to determine the cytotoxic effects of TQ-NLC on breast cancer (MDA-MB-231 and MCF-7) and cervical cancer cell lines (HeLa and SiHa). TQ-NLC was prepared by applying the hot high pressure homogenization technique. The mean particle size of TQ-NLC was 35.66 ± 0.1235?nm with a narrow polydispersity index (PDI) lower than 0.25. The zeta potential of TQ-NLC was greater than ?30?mV. Polysorbate 80 helps to increase the stability of TQ-NLC. Differential scanning calorimetry showed that TQ-NLC has a melting point of 56.73°C, which is lower than that of the bulk material. The encapsulation efficiency of TQ in TQ-NLC was 97.63 ± 0.1798% as determined by HPLC analysis. TQ-NLC exhibited antiproliferative activity towards all the cell lines in a dose-dependent manner which was most cytotoxic towards MDA-MB-231 cells. Cell shrinkage was noted following treatment of MDA-MB-231 cells with TQ-NLC with an increase of apoptotic cell population (P < 0.05). TQ-NLC also induced cell cycle arrest. TQ-NLC was most cytotoxic towards MDA-MB-231 cells. It induced apoptosis and cell cycle arrest in the cells. PMID:25632388

Ng, Wei Keat; Saiful Yazan, Latifah; Yap, Li Hua; Wan Nor Hafiza, Wan Abd Ghani; How, Chee Wun; Abdullah, Rasedee

2015-01-01

46

Thymoquinone-Loaded Nanostructured Lipid Carrier Exhibited Cytotoxicity towards Breast Cancer Cell Lines (MDA-MB-231 and MCF-7) and Cervical Cancer Cell Lines (HeLa and SiHa).  

Science.gov (United States)

Thymoquinone (TQ) has been shown to exhibit antitumor properties. Thymoquinone-loaded nanostructured lipid carrier (TQ-NLC) was developed to improve the bioavailability and cytotoxicity of TQ. This study was conducted to determine the cytotoxic effects of TQ-NLC on breast cancer (MDA-MB-231 and MCF-7) and cervical cancer cell lines (HeLa and SiHa). TQ-NLC was prepared by applying the hot high pressure homogenization technique. The mean particle size of TQ-NLC was 35.66 ± 0.1235?nm with a narrow polydispersity index (PDI) lower than 0.25. The zeta potential of TQ-NLC was greater than -30?mV. Polysorbate 80 helps to increase the stability of TQ-NLC. Differential scanning calorimetry showed that TQ-NLC has a melting point of 56.73°C, which is lower than that of the bulk material. The encapsulation efficiency of TQ in TQ-NLC was 97.63 ± 0.1798% as determined by HPLC analysis. TQ-NLC exhibited antiproliferative activity towards all the cell lines in a dose-dependent manner which was most cytotoxic towards MDA-MB-231 cells. Cell shrinkage was noted following treatment of MDA-MB-231 cells with TQ-NLC with an increase of apoptotic cell population (P < 0.05). TQ-NLC also induced cell cycle arrest. TQ-NLC was most cytotoxic towards MDA-MB-231 cells. It induced apoptosis and cell cycle arrest in the cells. PMID:25632388

Ng, Wei Keat; Saiful Yazan, Latifah; Yap, Li Hua; Wan Nor Hafiza, Wan Abd Ghani; How, Chee Wun; Abdullah, Rasedee

2015-01-01

47

Expression of LGI1 Impairs Proliferation and Survival of HeLa Cells  

OpenAIRE

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 significan...

Nadia Gabellini; Valentina Masola

2009-01-01

48

Nuclear blebbing of biologically active organoselenium compound towards human cervical cancer cell (HeLa): In vitro DNA/HSA binding, cleavage and cell imaging studies.  

Science.gov (United States)

New pharmacophore organoselenium compound (1) was designed, synthesized and characterized by various spectroscopic methods (IR, ESI-MS, (1)H, (13)C and (77)Se NMR) and further confirmed by X-ray crystallography. Compound 1 consists of two 3,5-bis(trifluoromethyl)phenyl units which are connected to the selenium atom via the organometallic C-Se bond. In vitro DNA binding studies of 1 was investigated by absorption and emission titration methods which revealed that 1 recognizes the minor groove of DNA in accordance with molecular docking studies with the DNA duplex. Gel electrophoretic assay demonstrates the ability of 1 to cleave pBR322 DNA through hydrolytic process which was further validated by T4 religation assay. To understand the drug-protein interaction of which ultimate molecular target was DNA, the affinity of 1 towards HSA was also investigated by the spectroscopic and molecular modeling techniques which showed hydrophobic interaction in the subdomain IIA of HSA. Furthermore, the intracellular localization of 1 was evidenced by cell imaging studies using HeLa cells. PMID:25535953

Rizvi, Masood Ahmad; Zaki, Mehvash; Afzal, Mohd; Mane, Manoj; Kumar, Manjeet; Shah, Bhahwal Ali; Srivastav, Saurabh; Srikrishna, Saripella; Peerzada, Ghulam Mustafa; Tabassum, Sartaj

2015-01-27

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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

50

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

Scientific Electronic Library Online (English)

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

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

51

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

52

Electron beam radiation dosage fixation on HeLa cell line  

International Nuclear Information System (INIS)

Cervical cancer continues to be a significant health burden worldwide. In the treatment of cancer radiation remains most frequently used important therapeutic model. The major drawback in radiotherapy is the development of radioresistant tumor cells, which will not effect by low dose radiation, one more important limitation is radiation induces normal tissue toxicity along with cancer cells. There is a need to find put solutions to these problems. HeLa is a cervical cancer cell line, which is epithelial cells of human cervix. Epithelial cells are known to be more radiosensitive than malignant tumors like gleoma (brain tumor). In our current study we exposed cultured HeLa cells to electron beam radiation to find out in vitro LD50 lethal dose and sublethal doses. Cultured HeLa cells were exposed to different doses of electron beam radiation 2 Gy, 4 Gy, 6 Gy, 8 Gy and 10 Gy. Cell viability was determined by MTI assay and Tryphan Blue assay, level of DNA damage was assessed using comet assay parameters. Our study revealed that as the radiation dose increased the number of viable cells decreased and percentage of DNA damage was increased. We conducted this study as a supportive study for our further assays. (author)

53

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

OpenAIRE

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

54

Tumoricidal effects of nanomaterials in HeLa cell line  

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The current study exhibits the cellular response of HeLa (cervical cancer) cells to metal oxides ultrafine nanomaterials e.g. manganese dioxide nanowires (MnO2 NRs), iron oxide nanoparticles (Fe2O3 NPs) and zinc oxide nanorods (ZnO NRs) as bare and as conjugated with photosensitizers. For cytotoxic evaluations, the cellular morphology, (MTT) assay, reactive oxygen species (ROS) production were used for cases with and without photo sensitizer as well illuminated with UV-visible laser exposed conditions. Three different photosensitizers were tested. These are 5-aminolevulinic acid (5-ALA), Photofrin® and protopor phyrin dimethyl ester (PPDME). Significant loss in cell viability was noted with 100-500 ?g/ml in bare and conjugated forms of the metal oxides used. The effect was insignificant with lower concentrations (0.05-50 ?g/ml). While notable anticancer effect of 5-ALA under 30 J/cm2 of diode laser irradiation was noted as compared to other photo sensitizer. By increasing the UV irradiation time of labeled cells, generation of ROS was observed, indicating the possibility of achieving efficient photodynamic therapy (PDT).

Fakhar-E-Alam, M.; Kishwar, S.; Khan, Y.; Siddique, M.; Atif, M.; Nur, O.; Willander, M.

2011-11-01

55

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

56

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

Science.gov (United States)

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

57

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

OpenAIRE

Introduction: Cervical cancer is the second prevalent cancer in developing countries and the sixth prevalent cancer in USA. Since conventional treatment methods are associated with detrimental side effects, searching for new drugs using natural ingredients is very important. Previous studies have shown that melittin (main component of honey bee venom) has anticancer properties along with the effect on cell membrane and activation of apoptosis. In this study, inhibitory effects of melittin on ...

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

2013-01-01

58

Antiproliferative effects of some medicinal plants on HeLa cells  

OpenAIRE

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

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

2013-01-01

59

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

60

Phosphatidylinositol anchor of HeLa cell alkaline phosphatase  

Energy Technology Data Exchange (ETDEWEB)

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

Jemmerson, R.; Low, M.G.

1987-09-08

61

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

Science.gov (United States)

Chrysin and its phosphate ester have previously been shown to inhibit cell proliferation and induce apoptosis in Hela cells; however, the underlying mechanism remains to be characterized. In the present study, we therefore synthesized diethyl flavon-7-yl phosphate (FP, C19H19O6P) by a simplified Atheron-Todd reaction, and explored its anti-tumor characteristics and mechanisms. Cell proliferation, cell cycle progression and apoptosis were measured by MTS, flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling techniques, respectively in human cervical cancer HeLa cells treated with 7-hydroxyflavone (HF) and FP. p21, proliferating cell nuclear antigen (PCNA) and cAMP levels in Hela cells were analyzed by western blot and radioimmunoassay. Both HF and FP inhibited proliferation and induced apoptosis in HeLa cells via induction of PCNA/p21 expression, cleaved caspase-3/poly (ADP-ribose) polymerase (PARP)-1, elevation of cAMP levels, and cell cycle arrest with accumulation of cells in the G0/G1 fraction. The effects of FP were more potent than those of HF. The interactions of FP with Ca2+-calmodulin (CaM) and Ca2+-CaM-phosphodiesterase (PDE)1 were explored by electrospray ionization-mass spectrometry and fluorescence spectra. FP, but not HF, formed non-covalent complexes with Ca2+-CaM-PDE1, indicating that FP is an inhibitor of PDE1, and resulting in elevated cellular cAMP levels. It is possible that the elevated cAMP levels inhibit growth and induce apoptosis in Hela cells through induction of p21 and cleaved caspase-3/PARP-1 expression, and causing down-regulation of PCNA and cell cycle arrest with accumulation of cells in the G0/G1 and G2/M fractions. In conclusion, FP was shown to be a Ca2+-CaM-PDE inhibitor, which might account for its underlying anti-cancer mechanism in HeLa cells. These observations clearly demonstrate the special roles of phosphorylated flavonoids in biological processes, and suggest that FP might represent a potential new drug for the therapy of human cervical carcinoma. PMID:22574207

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

2012-01-01

62

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

Science.gov (United States)

Chrysin and its phosphate ester have previously been shown to inhibit cell proliferation and induce apoptosis in Hela cells; however, the underlying mechanism remains to be characterized. In the present study, we therefore synthesized diethyl flavon-7-yl phosphate (FP, C(19)H(19)O(6)P) by a simplified Atheron-Todd reaction, and explored its anti-tumor characteristics and mechanisms. Cell proliferation, cell cycle progression and apoptosis were measured by MTS, flow cytometry and terminal deoxynucleotidyl transferase dUTP nick end labeling techniques, respectively in human cervical cancer HeLa cells treated with 7-hydroxyflavone (HF) and FP. p21, proliferating cell nuclear antigen (PCNA) and cAMP levels in Hela cells were analyzed by western blot and radioimmunoassay. Both HF and FP inhibited proliferation and induced apoptosis in HeLa cells via induction of PCNA/p21 expression, cleaved caspase-3/poly (ADP-ribose) polymerase (PARP)-1, elevation of cAMP levels, and cell cycle arrest with accumulation of cells in the G0/G1 fraction. The effects of FP were more potent than those of HF. The interactions of FP with Ca(2+)-calmodulin (CaM) and Ca(2+)-CaM-phosphodiesterase (PDE)1 were explored by electrospray ionization-mass spectrometry and fluorescence spectra. FP, but not HF, formed non-covalent complexes with Ca(2+)-CaM-PDE1, indicating that FP is an inhibitor of PDE1, and resulting in elevated cellular cAMP levels. It is possible that the elevated cAMP levels inhibit growth and induce apoptosis in Hela cells through induction of p21 and cleaved caspase-3/PARP-1 expression, and causing down-regulation of PCNA and cell cycle arrest with accumulation of cells in the G0/G1 and G2/M fractions. In conclusion, FP was shown to be a Ca(2+)-CaM-PDE inhibitor, which might account for its underlying anti-cancer mechanism in HeLa cells. These observations clearly demonstrate the special roles of phosphorylated flavonoids in biological processes, and suggest that FP might represent a potential new drug for the therapy of human cervical carcinoma. PMID:22574207

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

2012-01-01

63

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

64

Energy metabolism in hela and walker-256 cells  

International Nuclear Information System (INIS)

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

65

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

Scientific Electronic Library Online (English)

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

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

2012-04-01

66

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

International Nuclear Information System (INIS)

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

67

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

International Nuclear Information System (INIS)

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

68

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

Energy Technology Data Exchange (ETDEWEB)

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

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

2013-04-15

69

Berberine alters epigenetic modifications, disrupts microtubule network, and modulates HPV-18 E6-E7 oncoproteins by targeting p53 in cervical cancer cell HeLa: A mechanistic study including molecular docking.  

Science.gov (United States)

Increased evidence of chemo-resistance, toxicity and carcinogenicity necessitates search for alternative approaches for determining next generation cancer therapeutics and targets. We therefore tested the efficacy of plant alkaloid berberine on human papilloma virus (HPV) -18 positive cervical cancer cell HeLa systematically-involving certain cellular, viral and epigenetic factors. We observed disruptions of microtubule network and changes in membrane topology due to berberine influx through confocal and atomic force microscopies (AFM). We examined nuclear uptake, internucleosomal DNA damages, mitochondrial membrane potential (MMP) alterations and cell migration assays to validate possible mode of cell death events. Analytical data on interactions of berberine with pBR322 through fourier transform infrared (FTIR) and gel migration assay strengthen berberine?s biologically significant DNA binding abilities. We measured cellular uptake, DNA ploidy and DNA strand-breaks through fluorescence activated cell sorting (FACS). To elucidate epigenetic modifications, in support of DNA binding associated processes, if any, we conducted methylation-specific restriction enzyme (RE) assay, methylation specific-PCR (MSP) and expression studies of histone proteins. We also analyzed differential interactions and localization of cellular tumor suppressor p53 and viral oncoproteins HPV-18 E6-E7 through siRNA approach. We further made in-silico approaches to determine possible binding sites of berberine on histone proteins. Overall results indicated cellular uptake of berberine through cell membrane depolarization causing disruption of microtubule networks and its biological DNA binding abilities that probably contributed to epigenetic modifications. Results of modulation in p53 and viral oncoproteins HPV-18 E6-E7 by berberine further proved its potential as a promising chemotherapeutic agent in cervical cancer. PMID:25448308

Saha, Santu Kumar; Khuda-Bukhsh, Anisur Rahman

2014-12-01

70

Roscovitine-treated HeLa cells finalize autophagy later than apoptosis by downregulating Bcl?2.  

Science.gov (United States)

The cell cycle is tightly regulated by the family of cyclin-dependent kinases (CDKs). CDKs act as regulatory factors on serine and threonine residues by phosphorylating their substrates and cyclins. CDK?targeting drugs have previously demonstrated promising effects as cancer therapeutics both in vitro and in vivo. Roscovitine, a purine?derivative and specific CDK inhibitor, has been demonstrated to arrest the cell cycle and induce apoptosis in a number of different cancer cell lines, including HeLa cervical cancer cells. In the present study, roscovitine was able to decrease both the cell viability and cell survival as well as induce apoptosis in a dose?dependent manner in HeLa cells by modulating the mitochondrial membrane potential. The decrease of anti?apoptotic B-cell lymphoma 2 (Bcl?2) and Bcl-2 extra large protein expression was accompanied by the increase in pro?apoptotic Bcl-2-associated X protein and P53-upregulated modulator of apoptosis expression. The marked decrease in Bcl?2 following exposure to roscovitine (20 µM) for 48 h prompted us to determine the autophagic regulation. The outcome revealed that roscovitine triggered Beclin?1 downregulation and microtubule-associated light chain 3 cleavage starting from 12 h of incubation. Another biomarker of autophagy, p62, a crucial protein for autophagic vacuole formation, was diminished following 48 h. In addition, monodansyl cadaverin staining of autophagosomes also confirmed the autophagic regulation by roscovitine treatment. The expression levels of different Bcl?2 family members determined whether apoptosis or autophagy were induced following incubation with roscovitine for different time periods. Downregulation of pro?apoptotic Bcl?2 family members indicated induction of apoptosis, while the downregulation of anti?apoptotic Bcl?2 family members rapidly induced autophagosome formation in HeLa cells. PMID:25378060

Coker-Gurkan, Ajda; Arisan, Elif Damla; Obakan, Pinar; Ozfiliz, Pelin; Kose, Betsi; Bickici, Guven; Palavan-Unsal, Narcin

2015-03-01

71

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

International Nuclear Information System (INIS)

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

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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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Ascorbic acid: effects on ricin intoxicated HeLa cells  

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

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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

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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.

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

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

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

2013-10-01

78

The genomic and transcriptomic landscape of a HeLa cell line.  

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HeLa is the most widely used model cell line for studying human cellular and molecular biology. To date, no genomic reference for this cell line has been released, and experiments have relied on the human reference genome. Effective design and interpretation of molecular genetic studies performed using HeLa cells require accurate genomic information. Here we present a detailed genomic and transcriptomic characterization of a HeLa cell line. We performed DNA and RNA sequencing of a HeLa Kyoto cell line and analyzed its mutational portfolio and gene expression profile. Segmentation of the genome according to copy number revealed a remarkably high level of aneuploidy and numerous large structural variants at unprecedented resolution. Some of the extensive genomic rearrangements are indicative of catastrophic chromosome shattering, known as chromothripsis. Our analysis of the HeLa gene expression profile revealed that several pathways, including cell cycle and DNA repair, exhibit significantly different expression patterns from those in normal human tissues. Our results provide the first detailed account of genomic variants in the HeLa genome, yielding insight into their impact on gene expression and cellular function as well as their origins. This study underscores the importance of accounting for the strikingly aberrant characteristics of HeLa cells when designing and interpreting experiments, and has implications for the use of HeLa as a model of human biology. PMID:23550136

Landry, Jonathan J M; Pyl, Paul Theodor; Rausch, Tobias; Zichner, Thomas; Tekkedil, Manu M; Stütz, Adrian M; Jauch, Anna; Aiyar, Raeka S; Pau, Gregoire; Delhomme, Nicolas; Gagneur, Julien; Korbel, Jan O; Huber, Wolfgang; Steinmetz, Lars M

2013-08-01

79

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

International Nuclear Information System (INIS)

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 (P22 arrest induced by irradiation. (authors)

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Anticancer activity of certain herbs and spices on the cervical epithelial carcinoma (HeLa) cell line  

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

Danielle Berrington; Namrita Lall

2012-01-01

81

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

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

Rao, Shaival K; Rao, Priya S

2010-12-01

82

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)

83

In situ electrochemical assessment of cytotoxicity of chlorophenols in MCF-7 and HeLa cells.  

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An in situ electrochemical method was used to assess the cytotoxicity of chlorophenols using human breast cancer (MCF-7) and cervical carcinoma (HeLa) cells as models. On treatment with different chlorophenols, the electrochemical responses of the selected cells, resulting from the oxidation of guanine and xanthine in the cytoplasm, indicated the cell viability. In addition, the in situ in vitro electrochemical method was further compared with the traditional MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays. Although similar cytotoxicity data were obtained from both methods, the effective concentrations of chlorophenols that inhibited 50% cell growth (EC50 values) from the electrochemical method were only slightly lower than those from the MTT assay. These results indicate that the in situ in vitro electrochemical method paves a simple, rapid, strongly responsive, and label-free way to the cytotoxicity assessment of different chlorophenol pollutants. PMID:24973716

Qin, Hongwei; Liu, Jiguang; Zhang, Zeshi; Li, Jinlian; Gao, Guanggang; Yang, Yuxin; Yuan, Xing; Wu, Dongmei

2014-10-01

84

Doxorubicin-induced cell death requires cathepsin B in HeLa cells.  

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The cysteine protease cathepsin B acts as a key player in apoptosis. Cathepsin B-mediated cell death is induced by various stimuli such as ischemia, bile acids or TNF?. Whether cathepsin B can be influenced by anticancer drugs, however, has not been studied in detail. Here, we describe the modulation of doxorubicin-induced cell death by silencing of cathepsin B expression. Previously, it was shown that doxorubicin, in contrast to other drugs, selectively regulates expression and activity of cathepsin B. Selective silencing of cathepsin B by siRNA or the cathepsin B specific inhibitor CA074Me modified doxorubicin-mediated cell death in Hela tumor cells. Both Caspase 3 activation and PARP cleavage were significantly reduced in cells lacking cathepsin B. Moreover, mitochondrial membrane permeabilization as well as the release of cytochrome C and AIF from mitochondria into cytosol induced by doxorubicin were significantly diminished in cathepsin B suppressed cells. In addition, doxorubicin associated down-regulation of XIAP was not observed in cathepsin B silenced cells. Lack of cathepsin B significantly modified cell cycle regulatory proteins such as cdk1, Wee1 and p21 without significant changes in G(1), S or G(2)M cell cycle phases maybe indicating further cell cycle independent actions of these proteins. Consequently, cell viability following doxorubicin was significantly elevated in cells with cathepsin B silencing. In summary, our data strongly suggest a role of cathepsin B in doxorubicin-induced cell death. Therefore, increased expression of cathepsin B in various types of cancer can modify susceptibility towards doxorubicin. PMID:20709028

Bien, S; Rimmbach, C; Neumann, H; Niessen, J; Reimer, E; Ritter, C A; Rosskopf, D; Cinatl, J; Michaelis, M; Schroeder, H W S; Kroemer, H K

2010-11-15

85

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

86

A novel L1 retrotransposon marker for HeLa cell line identification  

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The HeLa cell line is the oldest, most widely distributed, permanent human cell line. As a nearly ubiquitous inhabitant of laboratories using tissue culture techniques, its aggressive growth characteristics make it a problematic contaminant that can overgrow less robust cell lines. Consequently, HeLa contamination is common in both the research laboratory and cell line repository contexts, and its detection is hampered by the lack of a rapid, sensitive and robust assay. Here we report the dev...

Rahbari, Raheleh; Sheahan, Tom; Modes, Vasileios; Collier, Pam; Macfarlane, Catriona; Badge, Richard M.

2009-01-01

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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

88

Leptomycin B increases radiosensitization by trichostain A in HeLa cells  

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

89

Leptomycin B increases radiosensitization by trichostain A in HeLa cells  

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

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

2005-06-15

90

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

91

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

International Nuclear Information System (INIS)

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

92

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

93

The Sensitivity of Hela Kyoto Cell Line Transfected with Sensor HyPer2 to Cisplatin  

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Full Text Available The aim of the investigation is to compare by means of MTT assay cytotoxic effect of cisplatin on the cells of HeLa Kyoto line and HeLa Kyoto line containing genetically-encoded sensor of hydrogen peroxide HyPer2 (HeLa Kyoto–HyPer2 line, and using staining by trypan blue to identify the doses of cisplatin causing cell death at different exposure time. Materials and Methods. A HeLa Kyoto cell line of human cervical carcinoma and HeLa Kyota line transfected with the cytoplasmic sensor of hydrogen peroxide (HeLa Kyoto–HyPer2 were used in the study. The analysis of cytotoxic and antiproliferative action of cisplatin in relation to the given cells was performed using MTT assay. Cell viability was determined after 24 h of incubation with the preparation at concentrations from 0 to 50 ?mol/L, then within the period from 0 to 24 h with an interval of 2 h at concentration of IC50; and also after 2, 4, 6, 8 h at concentrations from 9.3 to 833.3 ?mol/L a quantity of live and destructed cells was counted using staining by trypan blue. Results. After cisplatin expose the dose-response curves for cell viability of Hela Kyoto and HeLa Kyoto–HyPer2 cell lines were built according to MTT assay data. It was established that concentration of IC50 corresponding to the dose causing a loss of viability of 50% of cells is 1.3 times lower for HeLa Kyoto–HyPer2 compared to HeLa Kyoto. The results of staining by a vital agent trypan blue showed that inhibiting effects of cisplatin in concentration of IC50 by 24 h are mainly linked with the delay of cell division but not with their death. At concentrations up to 52 ?mol/L damage of the membranes does not occur during 8 h, and at superhigh concentrations — 416.7 ?mol/L — the damage is possible already 4 h after the exposure. Conclusion. Comparison of sensibility of the two cell lines to the effect of cisplatin showed that transfection of the cells with the fluorescent protein results in the increase of the sensitivity to cisplatin. When HeLa Kyoto–HyPer2 cells are exposed to the preparation at concentration of IC50 during 24 h, inhibition of cell division is observed; higher concentrations of the preparation cause increase of the number of dead cells and diminish the terms of their destruction.

A.S. Belova

2015-01-01

94

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

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

Syuhei Okamoto

2014-02-01

95

Mycoplasma fermentans binds to and invades HeLa cells: involvement of plasminogen and urokinase.  

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Adherence of Mycoplasma fermentans to HeLa cells followed saturation kinetics, required a divalent cation, and was enhanced by preincubation of the organism at 37 degrees C for 1 h in a low-osmolarity solution. Proteolytic digestion, choline phosphate, or anti-choline phosphate antibodies partially inhibited the adherence, supporting the notion that M. fermentans utilizes at least two surface components for adhesion, a protease-sensitive surface protein and a phosphocholine-containing glycolipid. Plasminogen binding to M. fermentans greatly increased the maximal adherence of the organism to HeLa cells. Anti-plasminogen antibodies and free plasminogen inhibited this increase. These observations suggest that in the presence of plasminogen the organism adheres to novel sites on the HeLa cell surface, which are apparently plasminogen receptors. Plasminogen-bound M. fermentans was detected exclusively on the cell surface of the infected HeLa cells. Nevertheless, plasminogen binding in the presence of the urokinase-type plasminogen activator (uPA) promoted the invasion of HeLa cells by M. fermentans. The latter finding indicates that the invasiveness of M. fermentans does not result from binding plasminogen but from activation of the bound plasminogen to plasmin. Cholesterol depletion and sequestration with beta-cyclodextrin and filipin, respectively, did not affect the capacity of M. fermentans to adhere, but invasion of HeLa cells by uPA-activated plasminogen-bound M. fermentans was impaired, suggesting that lipid rafts are implicated in M. fermentans entry. PMID:15321992

Yavlovich, Amichai; Katzenell, Avigail; Tarshis, Mark; Higazi, Abd A-R; Rottem, Shlomo

2004-09-01

96

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

97

Modification of radiation response in HeLa cells by misonidazole  

International Nuclear Information System (INIS)

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)

98

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

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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

99

Photodynamic damage study of HeLa cell line using ALA  

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The present study evaluates the photodynamic damage with 5-aminolevulinic acid (5-ALA) using HeLa as experimental model. HeLa cell line was irradiated with red light (He-Ne laser, ? = 632.8 CW nm). The influence of different incubation times and concentrations of 5-ALA, different irradiation doses and various combinations of photosensitizer and light doses on the cellular viability of HeLa cells were studied. The optimal uptake of photosensitizer ALA in HeLa cells was investigated by means of PpIX fluorescence intensity by exciting the HeLa cell suspension at 450 nm and a detection wavelength set at 690 nm. Cells viability was determined by means of trypan blue solution. The spectrometric measurements showed that the maximal cellular uptake of 5-ALA occurred after 4 h in vitro incubation. We found that the combination with 5-ALA and laser irradiation leads to time/concentration-dependent increase of cells death and also energy doses-dependent enlarge the cells death. The fluorescence intensity after PDD of carcinoma cells reduce when compared with the control group. The fluorescence emission spectral profiles after PDD of carcinoma cells showed a dip around 425-525 nm when compared with the control group. This may be due to the damage of mitochondria component of cells. The percentage of HeLa cells after PDD shows that the percentage of cells survival rate as function of laser dose (power). Hence it is clear that at 200 ?g/ml ALA and 20 mW laser irradiation, more than 70% of HeLa cells were dead after 15 min.

AlSalhi, M. S.; Atif, M.; AlObiadi, A. A.; Aldwayyan, A. S.

2011-04-01

100

Heterofucan from Sargassum filipendula Induces Apoptosis in HeLa Cells  

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

Hugo Alexandre Oliveira Rocha

2011-04-01

101

FRAKSINASI PROTEIN KAPANG LAUT Xylaria psidii KT30 DAN SITOTOKSISITASNYA TERHADAP SEL HeLa [Fractionation of Proteins of Marine Fungus Xylaria psidii KT30 and their Cytotoxicity against HeLa Cells  

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Full Text Available Cervical cancer is the most common cause of death for Indonesian women after human breast cancer. One of the efforts of cancer treatment is the utilization of natural compounds. One of the microorganisms having the potential as anticancer agent is endophytic fungi. Endophytic fungi from the marine habitat can be isolated from sea weeds, sea grasses, sponges, and mangroves. Xylaria psidii KT30, a marine fungus used in this study was isolated from red seaweed Kappaphycus alvarezii. Xylaria psidii KT30 was cultivated in potato dextrose broth medium for nine days at room temperature 27-29°C in shaking condition. This study aimed to obtain protein fractions from X. psidii KT30 and determine their toxicity againt Chang and HeLa cells. The fractionation process was conducted using DEAE Sephadex A-50 column chromatography and the toxicity was determined by Brine Shrimp Lethality Test (BSLT. The metabolites excreted in the culture broth was extracted using 90% of ammonium sulphate. The extract was then tested for their toxicity against HeLa and Chang cells by Microculture Tetrazolium Technique (MTT assay.The results revealed that LC50 of the protein extract of X. psidii KT30 was 104.95 ppm and IC50 was 69.9 ppm. Based on the National Cancer Institute (NCI, this value showed moderate cytotoxicity against HeLa cells.

Mita Gebriella Inthe

2014-06-01

102

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)

103

Adjuvant antiproliferative and cytotoxic effect of aloin in irradiated HeLaS3 cells  

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Naturally occurring phytoanthracycline, aloin, was used to radiosensitize HeLaS3 human cervix carcinoma cells. The results indicated that the cytotoxic adjuvant effect of aloin was synergistic with gammaionizing radiation at all drug concentrations and comparable to the cytotoxicity of 5-10 Gy ionizing radiation alone. Radiosensitization of HeLaS3 cells was achieved by 60 ?M aloin, which reduced the IC50 dose of ionizing radiation from 3.4 to 2 Gy. Ionizing radiation and aloin alone or in combination are shown to cause perturbation of the HeLaS3 cell-cycle and increase the percentage of cells in the DNA synthesis (S) phase of the cell cycle. While either of the agents applied alone causes programmed cell death by apoptosis, the simultaneous cell damage by both agents through the altered redox balance compromised cell capacity to conduct this program and led to synergic cytotoxic cell death by necrosis.

Ni?iforovi?, A.; Adži?, M.; Zari?, B.; Radoj?i?, M. B.

2007-09-01

104

Hyperthermia HeLa cell treatment with silica coated manganese oxide nanoparticles  

CERN Document Server

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

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

2009-01-01

105

[Lactobacillus inhibit adhesion of Staphylococcus aureus to HeLa cells].  

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To assess the ability of the previously selected human vaginal isolates of Lactobacillus crispatus (L. crispatus) T79-3, T90-1 and Lactobacillus jensenii (L. jensenii) T118-3, T231-1 to inhibit the growth of Staphylococcus aureus and block their adhesion to HeLa cells. The inhibitory bioactive substances produced by these Lactobacillus were also identified. Inhibitory substances interaction tests were carried out by using a streak-diffusion method on agar plates. Three types of interaction were performed to determine the inhibitory effect of Lactobacillus on adhesion of Staphylococcus aureus to HeLa cells: Exclusion Group (Lactobacillus and HeLa followed by pathogens), Competition Group (Lactobacillus, HeLa and pathogens together) and Displacement Group (pathogens and HeLa followed by the addition of Lactobacillus). The number of HeLa cells adhered to Staphylococcus aureus was quantified by bacteria colony counts on LB plate. The results showed that lactic acids produced by the Lactobacillus are the main substances that can inhibit Staphylococcus aureus growth and there is variation among the three types of interaction regarding the inhibitory activity against Staphylococcus aureus. The effects of Lactobacillus on blocking the adhesion to HeLa cells were concentration dependent. All four Lactobacillus isolates displayed the ability to inhibit Staphylococcus aureus growth and block Staphylococcus aureus adherence to HeLa cells. Exclusion Group was the most effective, and T79-3 showed greater capacity to block Staphylococcus aureus adherence compared with the other three isolates. The present study suggests the potential ability of L. crispatus T79-3 as probiotic for the treatment and prevention of urogenital infections in women. PMID:23016308

Wang, Jiang; Zhang, Ruifen; Zhou, Li; Su, Xiaohu; Hu, Chunhong; Zhu, Baoli; Feng, Tao

2012-06-01

106

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

107

Adjuvant antiproliferative and cytotoxic effect of aloin in irradiated HeLaS3 cells  

International Nuclear Information System (INIS)

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

108

ABT-737 Induces Bim Expression via JNK Signaling Pathway and Its Effect on the Radiation Sensitivity of HeLa Cells  

OpenAIRE

ABT-737 is a BH3 mimetic small molecule inhibitor that can effectively inhibit the activity of antiapoptotic Bcl-2 family proteins including Bcl2, Bcl-xL and Bcl-w, and further enhances the effect of apoptosis by activating the proapoptotic proteins (t-Bid, Bad, Bim). In this study, we demonstrate that ABT-737 improved the radiation sensitivity of cervical cancer HeLa cells and thereby provoked cell apoptosis. Our results show that ABT-737 inhibited HeLa cell proliferation and activated JNK a...

Wang, Huan; Yang, Yue-bo; Shen, Hui-min; Gu, Jian; Li, Tian; Li, Xiao-mao

2012-01-01

109

Adhesion and proliferation of HeLa and fibroblast cells on chemically-modified gold surfaces.  

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The development of materials that allow proper functioning of cells on solid supports is directly relevant to the construction of living-cell biosensors. Both physical and chemical properties of the surfaces have been shown to be critical in this field. Our aim is to report correlations between chemical properties of surfaces and cell behavior by studying adhesion, viability and proliferation of fibroblasts and HeLa cells. Neither fibroblasts nor HeLa cells adhered to a hydrophobic surface. Fibroblasts were able to attach and proliferate well on all other surfaces tested. In contrast, on some surfaces where HeLa cells adhered and were viable, proliferation decreased by half while on others proliferation was not affected. Proliferation was significantly correlated with the level of adsorption of serum proteins on the surface (quantified by surface plasmon resonance), but not with surface wettability (water contact angle). Interestingly, surfaces modified with COOH and HSO3 groups were the ones that favored most protein adsorption and allowed the best measures for HeLa cell proliferation. The decrease of HeLa cell proliferation on surfaces covered with poly-l-lysine (PL) was related with the profile of integrin expression. Compared to a polystyrene control surface, there was an increase in ?V and ?V?3 and a decrease in ?2 and ?3, indicating that migration rather than proliferation could be favored on PL functionalized surfaces. These results indicate that charge is more important than wettability to determine biocompatibility. PMID:25448718

Santos, Patricia A; Rocha, Cleidiane S; Baptista, Mauricio S

2014-11-01

110

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

111

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

OpenAIRE

The aim of this study is to investigate anticancer activity of methanol extract (ethylacetate, n-buthanol and water partitions) and water extract from Sarang semut (local name), Myrmecodya pendens which is one of Rubiaceae family. Within Papua area (Indonesia), this medicinal plant has been used traditionally as alternative treatment for ulcer, tumor and cancer. In this study, the extracts of this plant were tested for their activities in some cancer cells (HeLa and MCM-B2 cell)...

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

2010-01-01

112

Hyperthermia HeLa cell treatment with silica coated manganese oxide nanoparticles  

OpenAIRE

The effect of a high frequency alternating magnetic field on HeLa tumour cells incubated with ferromagnetic nanoparticles of manganese oxide perovskite La0.56(SrCa)0.22MnO3 have been studied. The particles were subjected to a size selection process and coated with silica to improve their biocompatibility. The control assays made with HeLa tumour cells showed that cell survival and growth rate were not affected by the particle internalization in cells, or by the electromagnet...

Villanueva, A.; La Presa, P.; Alonso, Jm; Rueda, T.; Martinez, A.; Crespo, P.; Morales, Mp; Fernandez, Ma Gonzalez; Valdes, J.; Rivero, G.

2009-01-01

113

Comparison of rhinovirus A infection in human primary epithelial and HeLa cells  

OpenAIRE

HeLa cells are used to study the life cycles of many different viruses, including the human rhinoviruses (HRV) in the family Picornaviridae. Although the natural targets of HRV are human bronchial epithelial cells (hBE), it is generally more difficult to obtain and maintain the relevant primary cell cultures, relative to HeLa cells. Given that the HRV are now identified as a major cause of human asthma exacerbations, it becomes important to document how much of the virus biology learned from ...

Amineva, S. P.; Aminev, A. G.; Gern, J. E.; Palmenberg, A. C.

2011-01-01

114

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

115

p150 ADAR1 isoform involved in maintenance of HeLa cell proliferation  

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Full Text Available Abstract Background RNA-specific adenosine deaminase ADAR1 is ubiquitously expressed in a variety of mammalian cells and tissues. Although its physiological importance in non-nervous tissues has been confirmed by analysis of null mutation phenotypes, few endogenous editing substrates have been identified in numerous peripheral tissues and biological function of ADAR1 has not been fully understood. Methods A conditional site-specific, ribozyme-based gene knock-down strategy was utilized to study the function of full-length isoform of ADAR1 (p150 protein in HeLa cell. Double-stable HeLa cell lines were developed by transfecting HeLa Tet-On cells with a pTRE-derived plasmid that can express a hammerhead ribozyme against mRNA of p150 ADAR1 isoform under induction condition. Semi-quantitative RT-PCR and Western blotting were performed to measure the expression of p150 in selected cell clones. Cell proliferation was evaluated by means of MTT assay and growth curve analysis. Cellular morphological changes were observed under light microscope. Flow Cytometry was used for cell cycle analysis. Growth rate of cell transplants in BALB/c nude mice was also investigated. Results Both HeLa cell proliferation in vitro and the growth rate of transplanted HeLa cell-derived tumors in nude mice in vivo were significantly inhibited due to reduced expression of ADAR1 p150. Additionally, cell cycle analysis showed that cell progression from G1 phase to S phase was retarded in the ADAR1 p150 suppressed cells. Conclusion Our results suggest that normal expression and functioning of p150 ADAR1 is essential for the maintenance of proper cell growth. The mechanisms underlying ADAR1's action might include both editing of currently unknown double-stranded RNAs and interacting with other cellular dsRNA-related processes.

Wu Yumei

2006-12-01

116

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

117

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

Science.gov (United States)

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

118

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

119

A novel trifluoromethyl benzopyran induces G1 cell cycle arrest and apoptosis in HeLa human cervical carcinoma cells.  

Science.gov (United States)

In the present study, a biologically active 4-(trifluoromethyl)phenyl piperazin moiety was linked to a 2,2- dimethyl -2H-benzopyran template to generate (3R,4S)-2,2-dimethyl-6-nitro-4-(4-(3-(trifluoromethyl)phenyl)piperazin-1-yl) chroman -3-ol (C110g), and the cellular and molecular mechanisms by which C110g exerts cytotoxic effects on the HeLa human cervical cancer cell line were further investigated. C110g suppressed the viability of HeLa cells in both concentration- and time-dependent manner (IC50 of 17 µM) by inducing DNA damage and G1 cell cycle arrest. Characteristic changes in nuclear morphology and Annexin V/PI staining pointed to apoptosis as the mode of cell death. The levels of p53 and p21 were increased in the C110g-treated cells, with a corresponding increase in Bax/Bcl-2 protein ratio. Subsequently, C110g induced the cytoplasmic release of cytochrome c from the mitochondria accompanied by a decreased mitochondrial membrane potential and activation of caspase-3 and -9. These results confirmed that the C110g transduced the apoptotic signal via the mitochondrial pathway. Caspase-8, typically associated with the initiation of the death receptor pathway, was activated, suggesting the extrinsic pathway might also be involved. However, C110g did not result in reactive oxygen species (ROS) generation. Taken together, these findings indicate that the DNA damage-dependent p53-regulated mitochondrial pathway as well as the extrinsic pathway play a crucial role in C110g-induced apoptosis, which provide a better understanding of the molecular mechanisms of trifluoromethyl benzopyrans in cervical cancer. PMID:23708884

Zhang, Xin; Hwang, Jiyoung; Jia, Xian; Shin, Dong-Soo; You, Song; Kim, Dong-Kyoo

2013-08-01

120

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

Science.gov (United States)

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

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

2014-06-01

121

Fragmentation and partitioning of the Golgi apparatus during mitosis in HeLa cells.  

OpenAIRE

Osmium impregnation was used to determine the number of Golgi apparatus in both interphase and mitotic HeLa cells. The number was found to increase substantially during mitosis to the point where random partitioning alone would explain the nearly equal numbers found in each daughter cell.

Lucocq, J. M.; Warren, G.

1987-01-01

122

Desensitization of histamine H1 receptor-mediated inositol phosphate production in HeLa cells.  

OpenAIRE

1. Histamine stimulated the accumulation of total [3H]-inositol phosphates (IPn) in control HeLa cells with an EC50 of 3.7 +/- 0.7 microM in the presence of 10 mM LiCl. The maximum response to histamine after 15 min incubation was 43 +/- 5% over basal accumulation and occurred at a concentration of 1 mM histamine. 2. The histamine-induced IPn production in HeLa cells was confirmed as H1 receptor-mediated, since the H1 antagonist mepyramine (10(-6) M) inhibited the histamine response (10(-4) M...

Bristow, D. R.; Zamani, M. R.

1993-01-01

123

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

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

Andrew R. Collins

2013-04-01

124

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

International Nuclear Information System (INIS)

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

125

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

OpenAIRE

Chrysin and its phosphate ester have previously been shown to inhibit cell proliferation and induce apoptosis in Hela cells; however, the underlying mechanism remains to be characterized. In the present study, we therefore synthesized diethyl flavon-7-yl phosphate (FP, C19H19O6P) by a simplified Atheron-Todd reaction, and explored its anti-tumor characteristics and mechanisms. Cell proliferation, cell cycle progression and apoptosis were measured by MTS, flow cytometry and terminal deoxynucle...

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

2012-01-01

126

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

127

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

128

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

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

Elgjo Kjell

2009-07-01

129

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

OpenAIRE

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...

Svalastog, Anna Lydia; Martinelli, Lucia

2013-01-01

130

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

Science.gov (United States)

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

Berrington, Danielle; Lall, Namrita

2012-01-01

131

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

132

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

Science.gov (United States)

MP [4-(3',3'-dimethylallyloxy)-5-methyl-6-methoxyphthalide] was obtained from liquid culture of Pestalotiopsis photiniae isolated from the Chinese Podocarpaceae plant Podocarpus macrophyllus. MP significantly inhibited the proliferation of HeLa tumor cell lines. After treatment with MP, characteristic apoptotic features such as DNA fragmentation and chromatin condensation were observed in DAPI-stained HeLa cells. Flow cytometry showed that MP induced G1 cell cycle arrest and apoptosis in a dose-dependent manner. Western blotting and real-time reverse transcription-polymerase chain reaction were used to investigate protein and mRNA expression. MP caused significant cell cycle arrest by upregulating the cyclin-dependent kinase inhibitor p27(KIP1) protein and p21(CIP1) 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, p21(CIP1), p16(INK4a) 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. PMID:23903687

Chen, C; Yang, R L

2013-08-01

133

Eupafolin, a flavonoid isolated from Artemisia princeps, induced apoptosis in human cervical adenocarcinoma HeLa cells.  

Science.gov (United States)

Although eupafolin, a flavone found in Artemisia princeps Pampanini, has been shown to inhibit the growth of several human cancer cells, its mode of action is poorly understood. In this study, we investigated the pro-apoptotic activities of eupafolin in human cervical carcinoma HeLa cells. It was found that eupafolin induced apoptosis in a dose-dependent manner, as evidenced by DNA fragmentation and the accumulation of positive cells for annexin V. In addition, eupafolin triggered the activations of caspases-3, -6, -7, -8, and -9 and the cleavages of their substrates, such as, poly (ADP-ribose) polymerase and lamin A/C. Furthermore, treatment with eupafolin resulted in a loss of mitochondrial membrane potential (DeltaPsi(m)), increased the release of cytochrome c to the cytosol, and altered the expression levels of B-cell lymphoma 2 (Bcl-2) family proteins. Interestingly, caspase-8, an initiator caspase, was activated after the loss of DeltaPsi(m) and the activations of caspases-3 and -9. Moreover, treatment with z-DEVD-fmk (a specific caspase-3 inhibitor) and the overexpression of Bcl-2 prevented eupafolin-stimulated caspase-8 activation. Altogether, these results suggest that the eupafolin-induced apoptosis in HeLa cells is mediated by caspase-dependent pathways, involving caspases-3, -9, and -8, which are initiated by the Bcl-2-dependent loss of DeltaPsi(m). PMID:20397191

Chung, Kyung-Sook; Choi, Jung-Hye; Back, Nam-In; Choi, Myung-Sook; Kang, Eun-Kyung; Chung, Hae-Gon; Jeong, Tae-Sook; Lee, Kyung-Tae

2010-09-01

134

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

135

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

136

Oxygen depletion speeds and simplifies diffusion in HeLa cells.  

Science.gov (United States)

Many cell types undergo a hypoxic response in the presence of low oxygen, which can lead to transcriptional, metabolic, and structural changes within the cell. Many biophysical studies to probe the localization and dynamics of single fluorescently labeled molecules in live cells either require or benefit from low-oxygen conditions. In this study, we examine how low-oxygen conditions alter the mobility of a series of plasma membrane proteins with a range of anchoring motifs in HeLa cells at 37°C. Under high-oxygen conditions, diffusion of all proteins is heterogeneous and confined. When oxygen is reduced with an enzymatic oxygen-scavenging system for ? 15 min, diffusion rates increase by > 2-fold, motion becomes unconfined on the timescales and distance scales investigated, and distributions of diffusion coefficients are remarkably consistent with those expected from Brownian motion. More subtle changes in protein mobility are observed in several other laboratory cell lines examined under both high- and low-oxygen conditions. Morphological changes and actin remodeling are observed in HeLa cells placed in a low-oxygen environment for 30 min, but changes are less apparent in the other cell types investigated. This suggests that changes in actin structure are responsible for increased diffusion in hypoxic HeLa cells, although superresolution localization measurements in chemically fixed cells indicate that membrane proteins do not colocalize with F-actin under either experimental condition. These studies emphasize the importance of controls in single-molecule imaging measurements, and indicate that acute response to low oxygen in HeLa cells leads to dramatic changes in plasma membrane structure. It is possible that these changes are either a cause or consequence of phenotypic changes in solid tumor cells associated with increased drug resistance and malignancy. PMID:25418168

Edwald, Elin; Stone, Matthew B; Gray, Erin M; Wu, Jing; Veatch, Sarah L

2014-10-21

137

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

OpenAIRE

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

Darnell, R. B.

1984-01-01

138

The Golgi and Endoplasmic Reticulum Remain Independent during Mitosis in HeLa Cells  

OpenAIRE

Partitioning of the mammalian Golgi apparatus during cell division involves disassembly at M-phase. Despite the importance of the disassembly/reassembly pathway in Golgi biogenesis, it remains unclear whether mitotic Golgi breakdown in vivo proceeds by direct vesiculation or involves fusion with the endoplasmic reticulum (ER). To test whether mitotic Golgi is fused with the ER, we compared the distribution of ER and Golgi proteins in interphase and mitotic HeLa cells b...

Jesch, Stephen A.; Linstedt, Adam D.

1998-01-01

139

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

OpenAIRE

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

140

Involvement of the Nrf2 pathway in the regulation of pterostilbene-induced apoptosis in HeLa cells via ER stress.  

Science.gov (United States)

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 < 0.05) was found with regard to the unique expression levels of Nrf2/ARE downstream target genes in HeLa cells undergoing late apoptosis, levels that were restored with 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

2014-01-01

141

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

Science.gov (United States)

The immortal HeLa cells case is an intriguing example of bio-objectification processes with great scientific, social, and symbolic impacts. These cells generate questions about representation, significance, and value of the exceptional, variety, individuality, and property. Of frightening (a lethal cancer) and emarginated (a black, poor woman) origins, with their ability to "contaminate" cultures and to "spread" into spaces for becoming of extraordinary value for human knowledge, well-being, and economy advancements, HeLa cells have represented humanity, and emphasized the importance of individual as a core concept of the personalized medicine. Starting from the process leading from HeLa "cells" to HeLa "bio-objects," we focus on their importance as high quality bio-specimen. We discuss the tension between phenomenological characteristic of fundamental biological research and the variety of material and methodologies in epidemiology and personalized medicine. The emerging methodologies and societal changes reflect present EU policies and lead toward a new paradigm of science. PMID:23986283

Svalastog, Anna Lydia; Martinelli, Lucia

2013-08-01

142

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

International Nuclear Information System (INIS)

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.

143

Adenovirus type 5 precursor terminal protein-expressing 293 and HeLa cell lines.  

OpenAIRE

HeLa and 293 cell lines that express biologically active adenovirus type 5 precursor terminal protein (pTP) have been made. The amount of pTP synthesized in these cell lines ranges from barely detectable to greater than that observed in cells infected with the wild-type virus. The pTP-expressing cell lines permit the growth of a temperature-sensitive terminal protein mutant virus sub100r at the nonpermissive temperature. A higher percentage of the stably transfected 293 cell lines expressed t...

Schaack, J.; Guo, X.; Ho, W. Y.; Karlok, M.; Chen, C.; Ornelles, D.

1995-01-01

144

Immunofluorescent staining of poly(ADP-ribose) in situ in HeLa cell chromosomes in the M phase.  

OpenAIRE

Randomly and synchronously growing HeLa cells were tested for poly(ADP-ribose) by direct and indirect immunofluorescent antibody techniques. Fluorescence of poly(ADP-ribose) was seen only in the nuclei of intact cells when the direct immunofluorescent antibody technique was used but in both the nuclei and cytoplasm when the indirect immunofluorescent antibody technique was used; fluorescence in the cytoplasm was nonspecific. When randomly or synchronously growing HeLa cells were fixed in acet...

Kanai, Y.; Tanuma, S.; Sugimura, T.

1981-01-01

145

Influences of ? rays combined with N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) on the biological effects of Hela cells  

International Nuclear Information System (INIS)

By measuring the survivals of HeLa cells, DNA strand breaks and the activity of O6-methylguanine DNA methyltransferase, the biological effect of ?-rays and MNNG on HeLa cells was investigated. It was observed that the combined exposure of ?-rays and MNNG increased the cell lethality, DNA strand breaks and the inactivation of the O6-methylguanine DNA methyltransferase. The decrease of the survival curve parameters D0, Dq shown a synergism between ?-radiation and MNNG

146

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

International Nuclear Information System (INIS)

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

147

Trypanosoma cruzi trypomastigotes induce cytoskeleton modifications during HeLa cell invasion  

Scientific Electronic Library Online (English)

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

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

1014-10-01

148

Mycoplasma fermentans Binds to and Invades HeLa Cells: Involvement of Plasminogen and Urokinase  

OpenAIRE

Adherence of Mycoplasma fermentans to HeLa cells followed saturation kinetics, required a divalent cation, and was enhanced by preincubation of the organism at 37°C for 1 h in a low-osmolarity solution. Proteolytic digestion, choline phosphate, or anti-choline phosphate antibodies partially inhibited the adherence, supporting the notion that M. fermentans utilizes at least two surface components for adhesion, a protease-sensitive surface protein and a phosphocholine-containing glycolipid. Pl...

Yavlovich, Amichai; Katzenell, Avigail; Tarshis, Mark; Higazi, Abd A. -r; Rottem, Shlomo

2004-01-01

149

Detection of Clostridium difficile toxin with McCoy cell monolayers and cell suspensions and comparison with HeLa cell assay.  

OpenAIRE

McCoy cell monolayers were compared with HeLa cell monolayers for the detection of Clostridium difficile toxin in 301 stool samples. Tests were positive (greater than or equal to 1/100 dilution) in 83 and 81 specimens tested with McCoy and HeLa cell monolayers, respectively. McCoy cell suspensions were compared with HeLa cell monolayers in 532 stool filtrates. Overall, 90 positive specimens were within one dilution and 432 filtrates were negative with either test, giving a correlation coeffic...

Maniar, A. C.; Chubb, H.; Louie, T. J.; Williams, T. W.; Forsyth, W.; Wilt, J. C.

1984-01-01

150

Purification of a HeLa cell receptor protein for group B coxsackieviruses.  

OpenAIRE

Coxsackievirus B3 (CB3) firmly attaches to HeLa cells, forming a specific complex between the virus and its receptor on the cell surface. We extracted this virus-receptor complex (VRC) with the detergents sodium deoxycholate and Triton X-100. The VRC was identified by its sedimentation coefficient (140S), which was less than that of virions (155S). Formation of the VRC from cell lysates and 3H-CB3 occurred with the same specificity as did attachment of virions to cells, in that its formation ...

Mapoles, J. E.; Krah, D. L.; Crowell, R. L.

1985-01-01

151

Deoxypodophyllotoxin induces G2/M cell cycle arrest and apoptosis in HeLa cells.  

Science.gov (United States)

The natural flavolignan deoxypodophyllotoxin (DPPT) inhibits tubulin polymerization and induces cell cycle arrest at G(2)/M, followed by apoptosis. However, the precise mechanism of DPPT action is currently unknown. Here, we investigated the mechanism by which DPPT treatment of HeLa cervical carcinoma cells induces cell cycle arrest and apoptosis. We show that DPPT treatment inhibits cell viability in a dose-dependent manner and that this reduction in cell viability results from cell cycle arrest at G(2)/M phase, accompanied by an increase in apoptotic cell death. The induction of apoptosis by DPPT was confirmed by visualization of morphologic changes and internucleosomal DNA fragmentation. In addition, DPPT causes p53 and Bax to accumulate, accompanied by activation of DNA damage-sensing kinases, including ataxia-telangiectasia mutated (ATM) kinase and Chk2. Furthermore, DPPT activates caspase-3 and -7, suggesting that caspase-mediated pathways are involved in DPPT-induced apoptosis. Levels of the tumor suppressor PTEN were up-regulated during DPPT treatment, coincident with Akt inhibition. Together, these data suggest that DPPT induces G(2)/M cell-cycle arrest followed by apoptosis through multiple cellular processes, involving the activation of ATM, upregulation of p53 and Bax, activation of caspase-3 and -7, and accumulation of PTEN resulting in the inhibition of the Akt pathway. PMID:19616373

Shin, Soon Young; Yong, Yeonjoong; Kim, Chang Gun; Lee, Young Han; Lim, Yoongho

2010-01-28

152

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

Energy Technology Data Exchange (ETDEWEB)

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

153

Total Phenolic and Flavonoid Contents of Aqueous Extract of Stinging Nettle and In Vitro Antiproliferative Effect on Hela and BT-474 Cell Lines.  

Science.gov (United States)

Phenolic compounds including flavonoids and phenolic acids are plants secondary metabolites. Due to their ability to act as antioxidant agents, there is a growing interest to use those components in traditional medicine for cancer prevention or treatment. The aim of this study was to measure the amounts of total phenolics and flavonoids as well as anti-proliferative effect of aqueous extract of Stinging nettle on BT-474 and Hela cell lines. The amounts of phenolics content and total flavonoids were determined by folin ciocalteu and aluminium chloride methods, respectively. The free radical scavenging activity was measured by using diphenyl - picrylhydrazyl (DPPH). The reducing power of the extract was measured in the presence of potassium hexacyanoferrate and its antiproliferative activity was assessed on BT-474 and Hela cell lines using MTT assay. Total phenolic content was 322.941± 11.811 mg gallic acid/g extract. Total flavonoid content was 133.916±12.006 mg Catechin/g. The IC50 of DPPH radical was 1.2 mg/ ml and the reducing power was 218.9± 15.582 ?g ascorbic acid/ g. Cell viability of BT-474 cells decreased to less than half of the control (no added extract) at the presence of 3 mg/ ml extract while no significant changes were detected for Hela cells at similar conditions. There was no significant difference in the percentage of surviving cells between consecutive days (day 1, 2 and 3) for both BT-474 and Hela cells (P>0.05). Although the relatively high amount of phenolic and flavonoid contents of the aqueous extract make this plant a promising candidate for diseases treatment; however, there is not a direct relationship between the amounts of these antioxidant components and the efficiency in in vitro cancer treatment. PMID:25035860

Fattahi, Sadegh; Zabihi, Ebrahim; Abedian, Zeinab; Pourbagher, Roghayeh; Motevalizadeh Ardekani, Ali; Mostafazadeh, Amrollah; Akhavan-Niaki, Haleh

2014-01-01

154

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

Science.gov (United States)

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

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

2014-01-01

155

Isolated HeLa cell nuclei synthesize meaningful DNA.  

OpenAIRE

DNA replicated at the beginning of S phase was labelled by incubating nuclei isolated from cells arrested at the G/S border with radioactive deoxyribonucleoside triphosphate in a reaction mixture sustaining DNA synthesis. By hybridization against ribosomal RNA bound to nitrocellulose, the fraction of the labelled DNA which was complementary to rRNA could be quantified, and the stability of the RNA-DNA hybrids could be estimated by sequential elution of DNA at increasing temperatures. The resu...

Kristensen, T.; Prydz, H.

1985-01-01

156

Laser stimulation can activate autophagy in HeLa cells  

International Nuclear Information System (INIS)

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.

157

Laser stimulation can activate autophagy in HeLa cells  

Science.gov (United States)

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

158

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

Directory of Open Access Journals (Sweden)

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

A. Soeksmanto

2010-01-01

159

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

Directory of Open Access Journals (Sweden)

Full Text Available To investigate the effects of N-(4-hydroxyphenyl retinamide (4HPR on the proliferation and apoptosis of Hela cells, the cell growth was observed by SRB test, colony-forming test and nude mice carcinogenic test. Morphologic changes of apoptosis were observed under microscopes and the normal, apoptotic and necrotic cells were identified by fluorescence staining. The biochemical features and percentage of apoptosis were performed by flow cytometry (FCM and DNA agarose gel electrophoresis. The results of SRB test, colony-forming test and nude mice carcinogenic test showed that 4HPR could inhibit the cells proliferation in vitro and in vivo (P<0.01. The apoptotic changes and apoptosis bodies could be observed under microscopes. The fluorescence staining revealed that 4HPR could induce the cells apoptosis, not necrosis. Typical DNA ladder appeared in 4HPR-treated cells and detected by FCM, the subdiploid nuclear peak appeared on the left of G1 peak and the apoptotic percentage was correlated with 4HPR in a dose and time dependent manner(P<0.01. All these results indicated that 4HPR could inhibit the proliferation of Hela cells and induce the cells apoptosis.

Xiao-Hong Han

2011-06-01

160

Retroviral expression of human cystatin genes in HeLa cells.  

Science.gov (United States)

Retroviral gene transfer is a highly efficient and effective method of stably introducing genetic material into the genome of specific cell types. The process involves the transfection of retroviral expression vectors into a packaging cell line, the isolation of viral particles, and the infection of target cell lines. Compared to traditional gene transfer methods such as liposome-mediated transfection, retroviral gene transfer allows for stable gene expression in cell populations without the need for lengthy selection and cloning procedures. This is particularly helpful when studying gene products that have negative effect on cell growth and viability. Here, we describe the retroviral transfer of cystatin cDNAs using HEK293-derived Phoenix packaging cells and human HeLa cervical carcinoma cells as target cells. PMID:25348302

Diep, Crystal M; Kaur, Gagandeep; Keppler, Daniel; Lin, Athena W

2015-01-01

161

Novel oxygenation system supports multilayer growth of HeLa cells.  

Science.gov (United States)

We have developed a novel substratum in which gelatin is bonded to a reservoir of perfluorodecalin using a perfluoroalkylating technique. This forms a stable substratum supporting good adhesion for cells. HeLa cells cultured on this substratum continued to grow exponentially after the surface was covered with a monolayer forming a tissue-like structure of more than 19 layers of cells. Histological sectioning and staining of the block of tissue formed revealed the presence of mitotic figures deep within the structure. Every cell was surrounded by other cells, similar to growth of cells in vivo. This technique opens up a new approach to studying problems involved in cell-cell interaction and development of histotypic structures in vitro. PMID:8891219

Rappaport, C; Trujillo, E M; Soong, L F

1996-10-01

162

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.

1994-06-01

163

Cell cycle-dependent subcellular distribution of ClC-3 in HeLa cells.  

Science.gov (United States)

Chloride channel-3 (ClC-3) is suggested to be a component and/or a regulator of the volume-activated Cl(-) channel in the plasma membrane. However, ClC-3 is predominantly located inside cells and the role of intracellular ClC-3 in tumor growth is unknown. In this study, we found that the subcellular distribution of endogenous ClC-3 varied in a cell cycle-dependent manner in HeLa cells. During interphase, ClC-3 was distributed throughout the cell and it accumulated at various positions in different stages. In early G1, ClC-3 was mainly located in the nucleus. In middle G1, ClC-3 gathered around the nuclear periphery as a ring. In late G1, ClC-3 moved back into the nucleus, where it remained throughout S phase. In G2, ClC-3 was concentrated in the cytoplasm. When cells progressed from G2 to the prophase of mitosis, ClC-3 from the cytoplasm translocated into the nucleus. During metaphase and anaphase, ClC-3 was distributed throughout the cell except for around the chromosomes and was aggregated at the spindle poles and in between two chromosomes, respectively. ClC-3 was then again concentrated in the nucleus upon the progression from telophase to cytokinesis. These results reveal a cell cycle-dependent change of the subcellular distribution of ClC-3 and strongly suggest that ClC-3 has nucleocytoplasmic shuttling dynamics that may play key regulatory roles during different stages of the cell cycle in tumor cells. PMID:22371056

Mao, Jianwen; Li, Xiaobo; Chen, Weiqiang; Xu, Bin; Zhang, Haifeng; Li, Hongzhi; Wang, Liwei; Jin, Xiaobao; Zhu, Jiayong; Lin, Guixian; Wang, Weizhang; Chen, Lixin

2012-06-01

164

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 is. 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)

165

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

OpenAIRE

BALB/c mice were immunized with HeLa cells, and their spleen cells were fused with myeloma cells to produce hybridomas. Initial screening of culture fluids from 800 fusion products in a cell protection assay against coxsackievirus B3 (CB3) and the CB3-RD virus variant yielded five presumptive monoclonal antibodies with three specificities: protection against CB3 on HeLa, protection against CB3-RD on rhabdomyosarcoma (RD) cells, and protection against both viruses on the respective cells. Only...

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

1986-01-01

166

Molecular characterization of an inwardly rectifying K+ channel from HeLa cells.  

Science.gov (United States)

Previous patch-clamp studies have shown that the potassium permeability of the plasma membrane in HeLa cells, a cell line derived from an epidermoid carcinoma of the cervix, is controlled by various K+-selective pores including an IRK1 type inwardly rectifying K+ channel. We used the sequence previously reported for the human heart Kir2.1 channel to design a RT-PCR strategy for cloning the IRK1 channel in HeLa cells. A full-length clone of 1.3 kb was obtained that was identical to the human cardiac Kir2.1 inward rectifier. The nature of the cloned channel was also confirmed in a Northern blot analysis where a signal of 5.3 kb corresponding to the molecular weight expected for a Kir2.1 channel transcript was identified not only in HeLa cells, but also in WI-38, ECV304 and bovine aortic endothelial cells. The HeLa IRK1 channel cDNA was subcloned in an expression vector (pMT21) and injected into Xenopus oocytes. Cell-attached and inside-out single channel recordings obtained from injected oocytes provided evidence for a voltage-independent K+-selective channel with current/voltage characteristics typical of a strong inward rectifier. The single channel conductance for inward currents measured in 200 mm K2SO4 conditions was estimated at 40 +/- 1 pS (n = 3), for applied voltages ranging from -100 to -160 mV, in agreement with the unitary conductance for the IRK1 channel identified in HeLa cells. In addition, the single channel conductance for inward currents, Gamma, was found to vary as a function of alphaK, the external K+ ion activity, according to Gamma = Gamma0 [alphaK]delta with Gamma0 = 3.3 pS and delta = 0.5. Single channel recordings from injected oocytes also provided evidence of a voltage-dependent block by external Cs+ and Ba2+. The presence of 500 micron Cs+ caused a voltage-dependent flickering, typical of a fast channel blocking process which resulted in a reduction of the channel open probability at increasingly negative membrane potential values. The fractional electrical distance computed for the Cs+ blocking site was greater than 1 indicating a multiple ion channel occupation. In contrast, external Ba2+ at concentrations ranging from 25 to 100 micron caused a slow channel block, consistent with the binding of a single Ba2+ ion at a site located at half the membrane span. It is concluded on the basis of these observations that HeLa cells expressed a Kir2.1 type inwardly rectifying channel likely to be involved in maintaining and regulating the cell resting potential. PMID:9878074

Klein, H; Garneau, L; Coady, M; Lemay, G; Lapointe, J Y; Sauvé, R

1999-01-01

167

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

DEFF Research Database (Denmark)

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

168

Biophysical characterization of gap-junction channels in HeLa cells.  

Science.gov (United States)

HeLa cells seem not to be junctionally coupled when probed with techniques such as Lucifer yellow spreading and/or ionic coupling measured with three inserted microelectrodes. When investigated with double whole-cell patch-clamp measurements, HeLa cells in monolayer cultures were electrically coupled in 39% of the cases with very low transjunctional conductances (average one to five open channels). These gap-junction channels had a single-channel conductance gamma = 26 +/- 6 pS and were voltage-gated with an equivalent gating charge z = 3.1 +/- 1.5 for a voltage of half-maximal inactivation Uo = 49 +/- 10 mV. The voltage-dependent component represents only 31 +/- 8% of the total junctional conductance. The voltage-insensitive conductance is characterized by a residual open probability po (infinity) = 0.34 +/- 0.12, which corresponds to a ratio Gmin/Gmax = 0.50 +/- 0.12. Dissociation of monolayer cells into cell pairs yielded about 58% coupled cell pairs with no notably altered single-channel properties. PMID:7692394

Eckert, R; Dunina-Barkovskaya, A; Hülser, D F

1993-08-01

169

Electroporation of micro-droplet encapsulated HeLa cells in oil phase.  

Science.gov (United States)

Electroporation (EP) is a method widely used to introduce foreign genes, drugs or dyes into cells by permeabilizing the plasma membrane with an external electric field. A variety of microfluidic EP devices have been reported so far. However, further integration of prior and posterior EP processes turns out to be very complicated, mainly due to the difficulty of developing an efficient method for precise manipulation of cells in microfluidics. In this study, by means of a T-junction structure within a delicate microfluidic device, we encapsulated HeLa cells in micro-droplet of poration medium in oil phase before EP, which has two advantages: (i) precise control of cell-encapsulating droplets in oil phase is much easier than the control of cell populations or individuals in aqueous buffers; (ii) this can minimize the electrochemical reactions on the electrodes. Finally, we successfully introduced fluorescent dyes into the micro-droplet encapsulated HeLa cells in oil phase. Our results reflected a novel way to realize the integrated biomicrofluidic system for EP. PMID:20803502

Xiao, Kang; Zhang, Mengying; Chen, Shuyu; Wang, Limu; Chang, Donald Choy; Wen, Weijia

2010-09-01

170

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)  

OpenAIRE

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 colorime...

Lukman, M.; RENY SYAHRUNI; MICHRUN NISA; AISYAH FATMAWATI

2014-01-01

171

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

Science.gov (United States)

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

172

Nuclear proteome analysis of cisplatin-treated HeLa cells  

Energy Technology Data Exchange (ETDEWEB)

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

Wu Wei [Department of Toxicology, Zhejiang University School of Public Health, 388 Yu-Hang-Tang Road, Hangzhou, Zhejiang 310058 (China); Institute of Hygiene, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang 310013 (China); Yan Chunlan; Gan Tieer; Chen Zhanghui [Department of Toxicology, Zhejiang University School of Public Health, 388 Yu-Hang-Tang Road, Hangzhou, Zhejiang 310058 (China); Lu Xianghong [Department of Pharmacy, Lishui People' s Hospital, Lishui, Zhejiang 323000 (China); Duerksen-Hughes, Penelope J. [Department of Basic Sciences, Division of Biochemistry, Loma Linda University School of Medicine, Loma Linda, CA 92354 (United States); Zhu Xinqiang, E-mail: zhuxq@zju.edu.cn [Department of Toxicology, Zhejiang University School of Public Health, 388 Yu-Hang-Tang Road, Hangzhou, Zhejiang 310058 (China); Yang Jun, E-mail: gastate@zju.edu.cn [Department of Toxicology, Zhejiang University School of Public Health, 388 Yu-Hang-Tang Road, Hangzhou, Zhejiang 310058 (China)

2010-09-10

173

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

International Nuclear Information System (INIS)

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

174

Hyperthermia Differently Affects Connexin43 Expression and Gap Junction Permeability in Skeletal Myoblasts and HeLa Cells  

Science.gov (United States)

Stress kinases can be activated by hyperthermia and modify the expression level and properties of membranous and intercellular channels. We examined the role of c-Jun NH2-terminal kinase (JNK) in hyperthermia-induced changes of connexin43 (Cx43) expression and permeability of Cx43 gap junctions (GJs) in the rabbit skeletal myoblasts (SkMs) and Cx43-EGFP transfected HeLa cells. Hyperthermia (42°C for 6?h) enhanced the activity of JNK and its target, the transcription factor c-Jun, in both SkMs and HeLa cells. In SkMs, hyperthermia caused a 3.2-fold increase in the total Cx43 protein level and enhanced the efficacy of GJ intercellular communication (GJIC). In striking contrast, hyperthermia reduced the total amount of Cx43 protein, the number of Cx43 channels in GJ plaques, the density of hemichannels in the cell membranes, and the efficiency of GJIC in HeLa cells. Both in SkMs and HeLa cells, these changes could be prevented by XG-102, a JNK inhibitor. In HeLa cells, the changes in Cx43 expression and GJIC under hyperthermic conditions were accompanied by JNK-dependent disorganization of actin cytoskeleton stress fibers while in SkMs, the actin cytoskeleton remained intact. These findings provide an attractive model to identify the regulatory players within signalosomes, which determine the cell-dependent outcomes of hyperthermia. PMID:25143668

Antanavi?i?t?, Ieva; Mildažien?, Vida; Stankevi?ius, Edgaras; Herdegen, Thomas; Skeberdis, Vytenis Arvydas

2014-01-01

175

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

International Nuclear Information System (INIS)

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)

176

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

International Nuclear Information System (INIS)

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

177

Structures of nuclear phosphoproteins characteristic of rapidly growing HeLa cells  

International Nuclear Information System (INIS)

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

178

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

OpenAIRE

Upon chromatographic fractionation of HeLa cell nuclear extracts, small RNAs of 145 and 66/65 nucleotides, respectively, were detected that are distinct from the abundant small RNAs present in the extract. These RNAs are precipitated by antibodies directed against the trimethylguanosine cap structure, characteristic for small nuclear RNAs (snRNAs) of the U type. The RNAs of 145 and 66/65 nucleotides appear to be associated with at least one of the proteins common to the major small nuclear ri...

Kra?mer, A.

1987-01-01

179

Methylation of DNA in HeLa cells after ultraviolet irradiation  

International Nuclear Information System (INIS)

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

180

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

Science.gov (United States)

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

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

2009-10-01

181

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

OpenAIRE

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 som...

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

1989-01-01

182

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

183

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

International Nuclear Information System (INIS)

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)

184

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

185

Small-interfering RNA-mediated silencing of the MAPK p42 gene induces dual effects in HeLa cells  

OpenAIRE

The genesis and progression of cervical cancer involve the mutation or deviant expression of numerous genes, including the activation of oncogenes (Ha-ras, C-myc, C-erbB2 and Bcl-2) and inactivation of tumor-suppressor genes (p53 and Rb). Previous studies showed that small-interfering RNAs (siRNAs) targeting the MAPK p42 gene partly inhibit proliferation and increase apoptosis in human cervical carcinoma HeLa cells. Results of a microarray analysis showed that MAPK p42 siRNA inhibited cell gr...

Yuan, Jing-yi; Liu, Li-ying; Wang, Pei; Li, Zong-fang; Ni, Lei; Wang, Aiying; Xiao, Sheng-xiang; Song, Tu-sheng; Huang, Chen

2010-01-01

186

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

187

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

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

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

1986-02-01

188

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

International Nuclear Information System (INIS)

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

189

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

Science.gov (United States)

BALB/c mice were immunized with HeLa cells, and their spleen cells were fused with myeloma cells to produce hybridomas. Initial screening of culture fluids from 800 fusion products in a cell protection assay against coxsackievirus B3 (CB3) and the CB3-RD virus variant yielded five presumptive monoclonal antibodies with three specificities: protection against CB3 on HeLa, protection against CB3-RD on rhabdomyosarcoma (RD) cells, and protection against both viruses on the respective cells. Only one of the monoclonal antibodies (with dual specificity) survived two subclonings and was studied in detail. The antibody was determined to have an immunoglobulin G2a isotype and protected cells by blockade of cellular receptors, since attachment of [35S]methionine-labeled CB3 was inhibited by greater than 90%. The monoclonal antibody protected HeLa cells against infection by CB1, CB3, CB5, echovirus 6, and coxsackievirus A21 and RD cells against CB1-RD, CB3-RD, and CB5-RD virus variants. The monoclonal antibody did not protect either cell type against 16 other immunotypes of picornaviruses. The monoclonal antibody produced only positive fluorescence on those cells which were protected against infection, and 125I-labeled antibody confirmed the specific binding to HeLa and RD cells. The results suggest that this monoclonal antibody possesses some of the receptor specificity of the group B coxsackieviruses. PMID:3003376

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

1986-02-01

190

Changes in cell cycle progression of HeLaS3 cells synchronized after X-ray irradiation  

International Nuclear Information System (INIS)

Objective: To observe the effect of different doses of X-rays on cell cycle progression of synchronized HeLaS3 cells. Methods: Using a double block method with thymidine and flow cytometric analysis, the changes in cell cycle progression of synchronized HeLaS3 cells were examined after 75 mGy and 2.0 Gy X-irradiation in G0/G1, S and G2 + M phases respectively, and the dose-response relationship given in G2/M phase was analyzed after irradiation. Results: The S and G2 phases occurred late 9-15 h after the releasing point regardless of whether HeLaS3 cells were irradiated with 2 Gy in G0/G1, S or G2 + M phases. HeLaS3 cells underwent a G2 arrest 9 and 12 h after the releasing point when 75 mGy irradiation was administered in G0/G1 and G2 + M phases, and this delay skipped completely at 12 and 15 h, respectively. Moreover, the cell cycle progression was accelerated. However, there was no G2 delay at 9 and 11 h when the cells were irradiated with 75 mGy in S phase, and in this case the cell cycle progression from G2/M to G0/G1 phase was accelerated. A study on the dose-effect relationship showed that the number of G0/G1 phase cells decreased, and G2 delayed after 0.025-2.0 Gy irradiation given in G2/M phase, and ation given in G2/M phase, and the delay was dose-dependent. However, the changes in the number of S phase cells were different between low (0.025-0.1 Gy) and higher (0.5-2.0 Gy) doses. Conclusion: G2 delay is a very radiosensitive parameter, which occurs in HeLaS3 cells after X-ray irradiation with a dose as low as 25 mGy when administered in G0/G1 and G2 + M phases, but this delay only occurs with doses above 0.1 Gy when the cells are irradiated in S phase

191

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

International Nuclear Information System (INIS)

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

192

Gefitinib inhibits the growth of Toxoplasma gondii in HeLa cells.  

Science.gov (United States)

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

193

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

Scientific Electronic Library Online (English)

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

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

2007-06-01

194

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

195

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

International Nuclear Information System (INIS)

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)

196

Phosphorylation of Smac by Akt promotes the caspase-3 activation during etoposide-induced apoptosis in HeLa cells.  

Science.gov (United States)

The Akt, family of serine/threonine protein kinases functions as key regulators of multiple aspects of cell behavior, such as survival, proliferation, migration, and carcinogenesis. Notably, Akt exerts its anti-apoptotic effects through the phosphorylation of numerous substrates related with cell cycle, genome stability, and cancer development. In this report, nevertheless, we focused our view on the novel role of Akt which involves in a pro-apoptotic action by phosphorylating second mitochondria derived activator of caspases (Smac) protein during etoposide-induced apoptotic processes. Our data reveals that Akt could bind to and phosphorylate Smac at serine residue 67, which enhances the ability of Smac to interact with the cytosolic X-chromosome linked IAP (XIAP) protein. The cellular interaction of wild-type Smac with XIAP was enhanced with similar activation kinetics of Akt activity, while this interaction was markedly attenuated in cells expressing the phosphorylation-defective mutant S67A-Smac during etoposide-induced apoptosis. Moreover, we provide the evidence indicating that the phosphorylation of Smac at ser-67 markedly upregulates the caspase-3 activity by promoting the interaction of Smac with XIAP. Taken together, we propose that the phosphorylation of Smac by Akt might be a novel mechanism that involves in amplification of caspase cascade pathway during etoposide-induced apoptosis in HeLa cells. © 2013 Wiley Periodicals, Inc. PMID:24038446

Jeong, Chul-Ho; Chun, Kyung-Soo; Kundu, Juthika; Park, Byoungduck

2015-02-01

197

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

OpenAIRE

MP [4-(3?,3?-dimethylallyloxy)-5-methyl-6-methoxyphthalide] was obtained from liquid culture of Pestalotiopsis photiniae isolated from the Chinese Podocarpaceae plant Podocarpus macrophyllus. MP significantly inhibited the proliferation of HeLa tumor cell lines. After treatment with MP, characteristic apoptotic features such as DNA fragmentation and chromatin condensation were observed in DAPI-stained HeLa cells. Flow cytometry showed that MP induced G1 cell cycle arrest...

Chen, C.; Yang, R. L.

2013-01-01

198

Effects of a tumor promoter on phospholipid metabolism in HeLa cells  

International Nuclear Information System (INIS)

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

199

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

Scientific Electronic Library Online (English)

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.

2011-10-01

200

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

Science.gov (United States)

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

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

2014-07-01

201

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

Science.gov (United States)

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

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

2013-05-01

202

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

203

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

International Nuclear Information System (INIS)

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)

204

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

DEFF Research Database (Denmark)

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

205

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

206

Evaluation of the antineoplastic activity of guduchi (Tinospora cordifolia) in cultured HeLa cells.  

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Exposure of HeLa cells to 0, 5, 10, 25, 50 and 100 microg/ml of guduchi extracts (methanol, aqueous and methylene chloride) resulted in a dose-dependent but significant increase in cell killing, when compared to non-drug-treated controls. The effects of methanol and aqueous extracts were almost identical. However, methylene chloride extract enhanced the cell killing effect by 2.8- and 6.8-fold when compared either to methanol or aqueous extract at 50 and 100 microg/ml, respectively. Conversely, the frequency of micronuclei increased in a concentration-dependent manner in guduchi-treated groups and this increase in the frequency of micronuclei was significantly higher than the non-drug-treated control cultures and also with respect to 5 microg/ml guduchi extract-treated cultures, at the rest of the concentrations evaluated. Furthermore, the micronuclei formation was higher in the methylene chloride extract-treated group than in the other two groups. The dose response relationship for all three extracts evaluated was linear quadratic. The effect of guduchi extracts was comparable or better than doxorubicin treatment. The micronuclei induction was correlated with the surviving fraction of cells and the correlation between cell survival and micronuclei induction was found to be linear quadratic. Our results demonstrate that guduchi killed the cells very effectively in vitro and deserves attention as an antineoplastic agent. PMID:9619860

Jagetia, G C; Nayak, V; Vidyasagar, M S

1998-05-15

207

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

208

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

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

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

2013-01-01

209

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

Science.gov (United States)

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

210

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

Energy Technology Data Exchange (ETDEWEB)

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

211

Haemagglutinins and adhesion of Salmonella typhimurium to HEp2 and HeLa cells.  

Science.gov (United States)

When fimbriate (Fim+) strains of Salmonella typhimurium were grown in static broth, many bacteria were in the fimbriate phase and bore fimbrial mannose-sensitive haemagglutinin (MSHA) that enabled them to adhere to guinea-pig and other erythrocytes and to agglutinate them in rocked tile and static settling tests. When either Fim+ or Fim- strains were grown on phosphate-buffered nutrient agar, the bacteria formed a diffusible, mannose-resistant haemagglutinin (MRHA) that gave dispersed sediments with sheep and pig erythrocytes in static settling tests, but without evidence of bacterial adhesion to the erythrocytes. On exposure, from above or from below, to cultured HEp2 and HeLa cells for 30 or 90 min at 37 degrees C, motile MSHA-rich, MRHA-negative broth-grown bacteria adhered to the cells in large numbers (e.g., 20-100/cell), but motile MSHA-negative, MRHA-negative broth-grown bacteria and non-motile MSHA-negative, MRHA-rich agar-grown bacteria adhered in only small numbers (usually less than 1/cell). Thus, strong adhesiveness of bacteria for cultured cells in vitro appears to depend upon the presence of MSHA, not MRHA, and as Fim- (MSHA-negative) strains of S. typhimurium are known to be highly infective in animals, a strong reaction in the in-vitro model does not reflect a property of the bacteria essential for infectivity in vivo. PMID:6135809

Tavendale, A; Jardine, C K; Old, D C; Duguid, J P

1983-08-01

212

N-methylation of the heterogeneous nuclear ribonucleoproteins in HeLa cells  

International Nuclear Information System (INIS)

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 thns (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

213

Purification of a HeLa cell receptor protein for group B coxsackieviruses.  

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Coxsackievirus B3 (CB3) firmly attaches to HeLa cells, forming a specific complex between the virus and its receptor on the cell surface. We extracted this virus-receptor complex (VRC) with the detergents sodium deoxycholate and Triton X-100. The VRC was identified by its sedimentation coefficient (140S), which was less than that of virions (155S). Formation of the VRC from cell lysates and 3H-CB3 occurred with the same specificity as did attachment of virions to cells, in that its formation was blocked by unlabeled CB3 but not by poliovirus. The VRC was purified 30,000-fold by differential and sucrose gradient centrifugation. Iodination with Na125I revealed that the purified VRC consisted of the normal CB3 proteins and one additional protein (RP-a) with an approximate molecular weight of 49,500. RP-a was eluted from the VRC and was shown to rebind with CB3 and CB1 virions but not with poliovirus type 1. We propose that Rp-a is a protein in the plasma membrane receptor complex which is responsible for the specific recognition and binding of the group B coxsackieviruses. PMID:2991580

Mapoles, J E; Krah, D L; Crowell, R L

1985-09-01

214

Recycling of surface sialoglycoconjugates in HTC and HeLa cells  

International Nuclear Information System (INIS)

Surface sialoglycoconjugates of HeLa and HTC cells were labeled with NaB[3H]4 after oxidation by NaIO4. The labeling procedure cleaves the sialic acids to a neuraminidase-sensitive 7-carbon derivative, 5-acetamido-3.5-dideoxy-L-arabino-heptulosonic acid, termed AcNeu7. After labeling, the radioactivity is lost from both cell types with biphasic kinetics. the half-time for the fast phase is about 4 to 5 h; the slow phase has a half-time of 100 to 200 h. About 30 h after labeling and at later times, approximately 30% of the cell-associated radioactivity is susceptible to removal by external neuraminidase, suggesting an exchange with an internal pool that is twice the size of the surface pool. An internal pool of relatively high specific activity compared to the surface was generated by labeling as above, following by a period of time to allow internalization and enzymatic removal of external neuraminidase-sensitive radioactivity. During subsequent reincubation in growth medium, the surface became relabeled from the internal pool, again reaching a 30% neuraminidase-sensitive plateau. The relabeling of the surface was confirmed by radioactivity measurements on isolated plasma membranes. [3H]AcNeu7 cannot be reutilized by these cells in the de novo membrane biosynthetic pathway. The argument is made that the labeled sialoglycoconjugates are recycling intact through the internal poolhe internal pool

215

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 ande 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)

216

Influences on time course of apoptosis of HeLaS3 cells after irradiation with different doses X rays  

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Changes of time course in apoptosis of HeLaS3 cells following irradiation with different doses X rays were observed. The percentages of apoptotic cells were examined by both flow cytometry (FCM) and image cytometry (ICM) after the cells were stained by prospidium iodine (PI) and Hoechst 33342 (HO). It was founded that the percentages of apoptotic cells increased after irradiation with 0.5-4.0 Gy X rays and the peak value emerged at the 12th hour after irradiation. In addition the values examined by both FCM and ICM were nearly same and the tendency was identical. So irradiation may induce apoptosis of HeLaS3 cell lines

217

Citotoxicidad en células hela de extractos de tres especies de plantas medicinales de Hidalgo, México / Cytotoxicity in hela cells from extracts of three medicinal plants species from Hidalgo, Mexico  

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Full Text Available SciELO Mexico | Language: Spanish Abstract in spanish Se evaluó la citotoxicidad en cultivos de células HeLa de los extractos etanólicos de tres especies de plantas, Juniperus dep-peana, Solanum rostratum y Bidens odorata, que se utilizan tradicionalmente en dos regiones del estado de Hidalgo, México, para el tratamiento de heridas, úlceras, tumores y [...] cáncer de matriz. La citotoxicidad más elevada la presentó el extracto de J. deppeana (CI50 = 4.63 µg/ml), el cual fue separado por cromatografía en placa de gel de sílice y la fracción principal (Rf = 0.28 ) mostró actividad citotóxica (CI50 = 0.79 µg/ ml). Aunque menor, el extracto de S. rostratum también presentó citotoxicidad (CI50 = 127.5 µg/ml). B. odorata fue inactiva. Abstract in english Ethanolic extracts of three medicinal plants, Juniperus deppeana, Solanum rostratum and Bidens odorata, which are used in folk medicine in Hidalgo, Mexico, for the treatment of wounds, ulcers, tumors and cancer, were tested in a HeLa cell line to evaluate their cytotoxic activity. The highest cytoto [...] xicity was found in the extract of J. deppeana (IC = 4.63 µg/ml); hence, this extract was separated via chromatography on a silica gel plate, from which the main fraction (Rf = 0.28) showed strong cyto-toxic activity (IC50 = 0.79 µg/ml). Whereas the extract of S. rostratum also exhibited cytotoxicity (IC50 = 127.5 µg/ml), that of B. odorata was inactive.

M.A., Villavicencio Nieto; B.E., Pérez Escandón; E., Mendoza Pérez; V., Maldonado Lagunas.

218

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 ofThese 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)

219

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

220

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

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

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

2011-05-13

221

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

Science.gov (United States)

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

222

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

223

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

224

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

225

Effects of ATPase inhibitors on the response of HeLa cells to Helicobacter pylori vacuolating toxin.  

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Approximately 50% of Helicobacter pylori strains produce a toxin in vitro that induces vacuolation of eukaryotic cells. To determine whether ion transport pathways are important in the formation of toxin-induced vacuoles, HeLa cells were incubated with H. pylori toxin in the presence of nine different inhibitors of ion-transporting ATPases. Oligomycin, an inhibitor of predominantly F1F0-type ATPases, had no effect on toxin activity. Inhibitors of predominantly V-type ATPases, exemplified by b...

Cover, T. L.; Reddy, L. Y.; Blaser, M. J.

1993-01-01

226

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 H2O2-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 H2O2 for 1 hour. DNA damage was significantly induced by H2O2 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

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

2012-01-01

227

Intracellular uptake and trafficking of difluoroboron dibenzoylmethane-polylactide nanoparticles in HeLa cells.  

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In this study, nanoparticles based on difluoroboron dibenzoylmethane-poly(lactic acid) (BF(2)dbmPLA) are prepared. Polylactic acid or polylactide is a commonly used degradable polymer, while the boron dye possesses a large extinction coefficient, high emission quantum yield, two-photon absorption, and sensitivity to the surrounding environment. BF(2)dbmPLA exhibits molecular-weight-dependent emission properties and can be formulated as stable nanoparticles, suggesting that its unique optical properties may be useful in multiple contexts for probing intracellular environments. Here we show that BF(2)dbmPLA nanoparticles are internalized into cultured HeLa cells by endocytosis, and that within the cellular milieu, they retain their fluorescence properties. BF(2)dbmPLA nanoparticles are photostable, resisting laser-induced photobleaching under conditions that destroy the fluorescence of a common photostable probe, LysoTracker Blue. Their endocytosis is also lipid-raft-dependent, as evidenced by their significant colocalization with cholera toxin B subunit in membrane compartments after uptake and their sensitivity of uptake to methyl-beta-cyclodextrin. Additionally, BF(2)dbmPLA nanoparticle endocytosis utilizes microtubules and actin filaments. Internalized BF(2)dbmPLA nanoparticles do not accumulate in acidic late endosomes and lysosomes but within a perinuclear nonlysosomal compartment. These findings demonstrate the feasibility of using novel BF(2)dbmPLA nanoparticles exhibiting diverse emission properties for in situ, live cell imaging and suggest that their endogenous uptake occurs through a lipid-raft-dependent endocytosis mechanism. PMID:20420413

Contreras, Janette; Xie, Jiansong; Chen, Yin Jie; Pei, Hua; Zhang, Guoqing; Fraser, Cassandra L; Hamm-Alvarez, Sarah F

2010-05-25

228

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

229

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

230

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

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

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

1984-07-15

231

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

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HeLa cells were implanted into athymic nude mice from tissue culture and solid tumors established (HeLa cell tumor or HCT). Large cell numbers of 1 X 107 were required to obtain consistent and progressive growth, and tumor growth followed a Gompertzian mode. Irradiation studies were carried out using acute Cobalt-60 (60Co), low-dose-rate (LDR) Cs-137 and LDR Cf-252. Cf-252, a neutron-emitting radioisotope, produced an immediate tumor shrinkage and regression response after a dose of 279 cGy. Acute 60Co or LDR Cs-137 irradiation with 1000 cGy had little effect on the HCT. After a dose of 2000 cGy of 60Co radiation tumor shrinkage followed a latent period of approximately 5 days. Cisplatin had no effect on the HCT in nude mice in stationary or late exponential growth

232

Acetylcholinesterase inhibition, antioxidant activity and toxicity of Peumus boldus water extracts on HeLa and Caco-2 cell lines.  

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This work aimed to study the inhibition on acetylcholinesterase activity (AChE), the antioxidant activity and the toxicity towards Caco-2 and HeLa cells of aqueous extracts of Peumus Boldus. An IC(50) value of 0.93 mg/mL, for AChE inhibition, and EC(50) of 18.7 ?g/mL, for the antioxidant activity, was determined. This activity can be attributed to glycosylated flavonoid derivatives detected, which were the main compounds, although boldine and other aporphine derivatives were also present. No changes in the chemical composition or the biochemical activities were found after gastrointestinal digestion. Toxicity of P. boldus decoction gave an IC(50) value 0.66 mg/mL for HeLa cells, which caused significant changes in the cell proteome profile. PMID:22617353

Falé, P L; Amaral, F; Amorim Madeira, P J; Sousa Silva, M; Florêncio, M H; Frazão, F N; Serralheiro, M L M

2012-08-01

233

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. 

Judong Luo

2013-03-01

234

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

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

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

2014-07-01

235

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

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

236

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

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

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

2009-12-01

237

Organellar proteome analyses of ricin toxin-treated HeLa cells.  

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Apoptosis triggered by ricin toxin (RT) has previously been associated with certain cellular organellar compartments, but the diversity in the composition of the organellar proteins remains unclear. Here, we applied a shotgun proteomics strategy to examine the differential expression of proteins in the mitochondria, nuclei, and cytoplasm of HeLa cells treated and not treated with RT. Data were combined with a global bioinformatics analysis and experimental confirmations. A total of 3107 proteins were identified. Bioinformatics predictors (Proteome Analyst, WoLF PSORT, TargetP, MitoPred, Nucleo, MultiLoc, and k-nearest neighbor) and a Bayesian model that integrated these predictors were used to predict the locations of 1349 distinct organellar proteins. Our data indicate that the Bayesian model was more efficient than the individual implementation of these predictors. Additionally, a Biomolecular Interaction Network (BIN) analysis was used to identify 149 BIN subnetworks. Our experimental confirmations indicate that certain apoptosis-related proteins (e.g. cytochrome c, enolase, lamin B, Bax, and Drp1) were found to be translocated and had variable expression levels. These results provide new insights for the systematic understanding of RT-induced apoptosis responses. PMID:25227225

Liao, Peng; Li, Yunhu; Li, Hongyang; Liu, Wensen

2014-09-16

238

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

239

Inhibiting PI3K/Akt pathway increases DNA damage of cervical carcinoma HeLa cells by drug radiosensitization.  

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This study examined the role of PI3K/Akt pathway in radiosensitization of DNA damage of cervical carcinoma cells. The 50% inhibition concentration (IC50) of cisplatin and docetaxel in HeLa cells was detected by Mono-nuclear cell direct cytotoxicity assay (MTT) in vitro. HeLa cells were treated by cisplatin/docetaxel of 10 percent of IC20 alone or combined with LY294002 for 24 h, and then radiated by different doses of X-ray. The cell survival ratio was obtained by means of clone formation. One-hit multi-target model was fitted to the cell survival curve to calculate dose quasithreshold (Dq), mean lethal dose (D0), 2Gy survival fraction (SF2) and sensitization enhancement ratio (SER). The pAkt and total Akt expression was detected by Western blotting and DNA damage by neutro-comet electrophoresis. The HeLa cells were randomly divided into 7 groups in terms of different treatments: Control; radiation treatment (RT) group; LY294002+RT group; cisplatin+RT group; docetaxel+RT group; LY294002+cisplatin+RT group; LY294002+docetaxel+RT group. The apoptosis ratio of each group was measured by flow cytometry. The results showed that docetaxel and cisplatin significantly enhanced the phosphorylation of Akt in radiation-treated HeLa cells. The Dq, D0 and SF2 in LY294002-contained groups were lower than those in docetaxel or cisplatin+RT group. The SER in the LY294002+docetaxel+RT group was 1.35 times that of the docetaxel+RT group, and that in the LY294002+cisplatin+RT group 1.26 times that of the cisplatin+RT group. The Comet electrophoresis showed that tail distance in the LY294002+cisplatin+RT group or LY294002+docetaxel+ RT group was longer than in the cisplatin+RT group or docetaxel+RT group. The apoptosis ratio in the LY294002+cisplatin+RT group or LY294002+docetaxel +RT group was higher than in the cisplatin+RT group or docetaxel+RT group. It was concluded that inhibiting PI3K/Akt pathway can increase the effect of docetaxel and cisplatin on the radiosensitivity of HeLa cells and DNA damage resulted from radiation. PMID:20556582

Xia, Shu; Yu, Shiying; Fu, Qiang; Liu, Fei; Zheng, Wei; Fu, Xiugen; Zhao, Yin

2010-06-01

240

Nuclear proteome analysis of benzo(a)pyrene-treated HeLa cells  

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Previously, we employed a proteomics-based 2-D gel electrophoresis assay to show that exposure to 10 ?M benzo(a)pyrene (BaP) during a 24 h frame can lead to changes in nuclear protein expression and alternative splicing. To further expand our knowledge about the DNA damage response (DDR) induced by BaP, we investigated the nuclear protein expression profiles in HeLa cells treated with different concentrations of BaP (0.1, 1, and 10 ?M) using this proteomics-based 2-D gel electrophoresis assay. We found 125 differentially expressed proteins in BaP-treated cells compared to control cells. Among them, 79 (63.2%) were down-regulated, 46 (36.8%) were up-regulated; 8 showed changes in the 1 ?M and 10 ?M BaP-treated groups, 2 in the 0.1 ?M and 10 ?M BaP-treated groups, 4 in the 0.1 ?M and 1 ?M BaP-treated groups, and only one showed changes in all three groups. Fifty protein spots were chosen for liquid chromatography–tandem mass spectrometry (LC–MS/MS) identification, and of these, 39 were identified, including subunits of the 26S proteasome and Annexin A1. The functions of some identified proteins were further examined and the results showed that they might be involved in BaP-induced DDR. Taken together, these data indicate that proteomics is a valuable approach in the study of environmental chemical–host interactions, and the identified proteins could provide new leads for better understanding BaP-induced mutagenesis and carcinogenesis.nesis.

241

Nuclear proteome analysis of benzo(a)pyrene-treated HeLa cells  

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Previously, we employed a proteomics-based 2-D gel electrophoresis assay to show that exposure to 10 {mu}M benzo(a)pyrene (BaP) during a 24 h frame can lead to changes in nuclear protein expression and alternative splicing. To further expand our knowledge about the DNA damage response (DDR) induced by BaP, we investigated the nuclear protein expression profiles in HeLa cells treated with different concentrations of BaP (0.1, 1, and 10 {mu}M) using this proteomics-based 2-D gel electrophoresis assay. We found 125 differentially expressed proteins in BaP-treated cells compared to control cells. Among them, 79 (63.2%) were down-regulated, 46 (36.8%) were up-regulated; 8 showed changes in the 1 {mu}M and 10 {mu}M BaP-treated groups, 2 in the 0.1 {mu}M and 10 {mu}M BaP-treated groups, 4 in the 0.1 {mu}M and 1 {mu}M BaP-treated groups, and only one showed changes in all three groups. Fifty protein spots were chosen for liquid chromatography-tandem mass spectrometry (LC-MS/MS) identification, and of these, 39 were identified, including subunits of the 26S proteasome and Annexin A1. The functions of some identified proteins were further examined and the results showed that they might be involved in BaP-induced DDR. Taken together, these data indicate that proteomics is a valuable approach in the study of environmental chemical-host interactions, and the identified proteins could provide new leads for better understanding BaP-induced mutagenesis and carcinogenesis.

Yan Chunlan; Chen Zhaojun; Li Huanrong; Zhang Guanglin [The First Affiliated Hospital, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310003 (China); Department of Toxicology, Zhejiang University School of Public Health, Hangzhou, Zhejiang 310058 (China); Li Feng [The First Renmin Hospital, Houma, Shanxi 043000 (China); Duerksen-Hughes, Penelope J. [Department of Basic Sciences, Loma Linda University School of Medicine, Loma Linda, CA 92354 (United States); Zhu Xinqiang, E-mail: zhuxq@zju.edu.cn [Department of Toxicology, Zhejiang University School of Public Health, Hangzhou, Zhejiang 310058 (China); Yang Jun, E-mail: gastate@zju.edu.cn [The First Affiliated Hospital, State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310003 (China); Department of Toxicology, Hangzhou Normal University School of Public Health, Hangzhou, Zhejiang 310036 (China)

2012-03-01

242

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

OpenAIRE

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...

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

2003-01-01

243

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

244

Lipoxin A4 Suppresses Lipopolysaccharide-Induced Hela Cell Proliferation and Migration via NF-?B Pathway.  

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Uterine cervical carcinoma (UCC) is one of the most common malignant tumors in females, and UCC has a close relationship with chronic cervicitis. As the endogenous "braking signal," lipoxins can regulate anti-inflammation and the resolution of inflammation. We investigated the effect of lipoxin A4 (LXA4) on the proliferation, apoptosis, and migration in lipopolysaccharide (LPS)-stimulated Hela cells. We demonstrated that LXA4 could significantly suppress p53, cyclin D1 expression, and migration of LPS-stimulated Hela cells via nuclear factor-?B (NF-?B) pathway, and these effects could be blocked by Boc-2, the specific inhibitor of FPR2/ALX (the receptor of LXA4). We presented evidence for a novel role of LXA4 on the proliferation and migration in LPS-stimulated Hela cells, and FPR2/ALX was involved in the procedures. These results showed that LXA4 could be a possible candidate for UCC therapy, and blocking the activation of NF-?B would be an effective drug target. PMID:25348861

Hao, Hua; Xu, Fen; Hao, Jian; He, Yuan-Qiao; Zhou, Xiao-Yan; Dai, Hua; Wu, Li-Qing; Liu, Fan-Rong

2014-10-28

245

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

246

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

247

In vitro study of 5-aminolevulinic acid-based photodynamic therapy for apoptosis in human cervical HeLa cell line  

International Nuclear Information System (INIS)

5-aminolevulanic acid (ALA), belonging among the promising second generation of sensitizers, was evaluated as an inducer of photodamage on HeLa (human cervical adenocarcinoma) cell line. A diode laser (635 nm) was used as a source for initiation of the photodynamic effect. We studied the influence of different incubation times, various concentrations of sensitizer, different irradiation doses and various combinations of sensitizer and light doses on the photodamage of HeLa cells. Viability of cells was determined by means of neutral red assay. The quantitative cellular uptake of ALA sensitizer was done by spectrophotometric measurements. No prominent cytotoxic or phototoxic effects on HeLa were observed due to sensitizer or light doses when studied independently of each other. However phototoxicity evoked by laser irradiated sensitizer was detected in HeLa cell line

248

Lung cancer - small cell  

Science.gov (United States)

Cancer - lung - small cell; Small cell lung cancer; SCLC ... About 15% of all lung cancer cases are SCLC. Small cell lung cancer is slightly more common in men than women. Almost all cases of SCLC are ...

249

Expression of amplified DNA sequences for ornithine transcarbamylase in HeLa cells: arginine residues may be required for mitochondrial import of enzyme precursor  

OpenAIRE

Expression of ornithine transcarbamylase (OTC), a nuclear-coded mitochondrial enzyme, was programmed in HeLa cells by the use of a strategy of gene co-amplification. HeLa cells, ordinarily devoid of OTC activity, were transfected with a plasmid containing viral regulatory elements joined with two cDNA sequences, one encoding the human OTC precursor and a second encoding a mutant mouse dihydrofolate reductase. After transfection and selection in increasing concentrations of methotrexate, sever...

1985-01-01

250

Paclitaxel-resistant HeLa cells have up-regulated levels of reactive oxygen species and increased expression of taxol resistance gene 1.  

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This study is to establish a paclitaxel (PTX)-resistant human cervical carcinoma HeLa cell line (HeLa/PTX) and to investigate its redox characteristics and the expression of taxol resistance gene 1 (Txr1). HeLa cells were treated with PTX and effects of PTX on cell proliferation were detected through cell counting and the MTT assay. Levels of cellular reactive oxygen species (ROS), reduced glutathione (GSH), and oxidized glutathione (GSSG) as well as the ratio of GSH to GSSG were measured by the 2,7-difluorescein diacetate (DCFH-DA) method and the 5,5'dithiobis(2-nitrobenzoic acid) (DTNB) method. Activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were determined by the nitrite formation method, the molybdate colorimetric method, and the DTNB colorimetric method, respectively. The level of Txr1 mRNA was determined by real-time PCR. Compared with the regular HeLa cells, HeLa/PTX cells were larger in size and had more cytoplasmic granules. The population doubling time for HeLa/PTX cells was 1.32 times of that of HeLa cells (P<0.01). HeLa/PTX cells showed stronger resistance to PTX than HeLa cells with a resistance index of 122.69. HeLa/PTX cells had higher levels of ROS (P<0.01) and Txr1 mRNA (P<0.01), lower level of GSH (P < 0.05), and lower activities of SOD (P<0.01) and GPx (P < 0.05) than HeLa cells. HeLa/PTX cells, with higher levels of ROS and Txr1 mRNA expression, are more resistant to PTX than HeLa cells. PMID:25015454

Bi, Wenxiang; Wang, Yuxia; Sun, Gaoying; Zhang, Xiaojin; Wei, Yongqing; Li, Lu; Wang, Xiaoyuan

2014-07-01

251

3-(3-Hydroxy-4-methoxyphenyl)-4-(3,4,5-trimethoxyphenyl)-1,2,5-selenadiazole (G-1103), a novel combretastatin A-4 analog, induces G2/M arrest and apoptosis by disrupting tubulin polymerization in human cervical HeLa cells and fibrosarcoma HT-1080 cells.  

Science.gov (United States)

Microtubule is a popular target for anticancer drugs. In this study, we describe the effect 3-(3-hydroxy-4-methoxyphenyl)-4-(3,4,5-trimethoxyphenyl)-1,2,5-selenadiazole (G-1103), a newly synthesized analog of combretastatin A-4 (CA-4), showing a strong time- and dose-dependent anti-proliferative effect on human cervical cancer HeLa cells and human fibrosarcoma HT-1080 cells. We demonstrated that the growth inhibitory effects of G-1103 in HeLa and HT-1080 cells were associated with microtubule depolymerization and proved that G-1103 acted as microtubule destabilizing agent. Furthermore, cell cycle analysis revealed that G-1103 treatment resulted in cell cycle arrest at the G2/M phase in a time-dependent manner with subsequent apoptosis induction. Western blot analysis revealed that down-regulation of cdc25c and up-regulation of cyclin B1 was related with G2/M arrest in HeLa and HT-1080 cells treatment with G-1103. In addition, G-1103 induced HeLa cell apoptosis by up-regulating cleaved caspase-3, Fas, cleaved caspase-8 expression, which indicated that G-1103 induced HeLa cell apoptosis was mainly associated with death receptor pathway. However, G-1103 induced HT-1080 cell apoptosis by up-regulating cleaved caspase-3, Fas, cleaved caspase-8, Bax and cleaved caspase-9 expression and down-regulating anti-apoptotic protein Bcl-2 expression, which indicated that G-1103 induced HT-1080 cell apoptosis was associated with both mitochondrial and death receptor pathway. Taken together, all the data demonstrated that G-1103 exhibited its antitumor activity through disrupting the microtubule assembly, causing cell cycle arrest and consequently inducing apoptosis in HeLa and HT-1080 cells. Therefore, the novel compound G-1103 is a promising microtubule inhibitor that has great potentials for therapeutic treatment of various malignancies. PMID:25529822

Zuo, Daiying; Guo, Dandan; Jiang, Xuewei; Guan, Qi; Qi, Huan; Xu, Jingwen; Li, Zengqiang; Yang, Fushan; Zhang, Weige; Wu, Yingliang

2015-02-01

252

Lithium thiazolidine-4-carboxylate: Synthesis, spectroscopic characterization and preliminary in vitro cytotoxic studies in human HeLa cells  

Science.gov (United States)

A new water-soluble lithium salt of thiazolidine-4-carboxylic acid was synthesized and characterized by chemical and spectroscopic techniques. Elemental and mass spectrometric (ESI-MS) analyses of the solid compound fit to the composition LiC 4H 6NSO 2. 1H, 13C nuclear magnetic resonance (NMR), [ 1H- 15N] NMR and infrared (IR) analyses permitted to elucidate the structure of the compound. Biological activity was evaluated by cytotoxic analysis using HeLa cells. Determination of cell death was assessed using a tetrazolium salt colorimetric assay, which reflects the cells viability.

Corbi, Pedro P.; Andrade, Fabiana C.; Massabni, Antonio C.; Heinrich, Tassiele A.; Souza, Pedro P. C.; Costa-Neto, Cláudio M.

2008-12-01

253

Two new neolignans from Patrinia scabra with potent cytotoxic activity against HeLa and MNK-45 cells.  

Science.gov (United States)

Two new neolignans, patrineolignan A (1) and patrineolignan B (2), together with seven known lignans, were isolated from the 90 % aqueous EtOH extract of the roots of Patrinia scabra. Their structures were elucidated on the basis of spectroscopic data (HRESIMS, IR, 1D and 2D NMR) and comparison with literature data. The two new neolignans were evaluated in vitro for cytotoxic properties against human cervical carcinoma HeLa cell line and gastric carcinoma MNK-45 cell line using the microculture tetrazolium assay, and both 1 and 2 exhibited strongly cytotoxic activity against the two tumor cell lines. PMID:23737105

Di, Lei; Yan, Guo-Qing; Wang, Ling-Yu; Ma, Wei; Wang, Kai-Jin; Li, Ning

2013-10-01

254

Differential suppression of the tumorigenicity of HeLa and SiHa cells by adeno-associated virus.  

OpenAIRE

Adeno-associated virus (AAV) is well known for suppression of oncogenesis in rodents, but its inhibitory effects on human carcinoma are less well understood. We report the differential ability of AAV to inhibit the tumorigenicity of two human cervical carcinoma cell lines. The wild-type AAV-2 DNA carried by a pSV2Neo vector was transfected into HeLa cells, which contain 50 copies of human papillomavirus type 18 (HPV-18), and SiHa cells, which contain 1-2 copies of HPV-16. About 1-3 copies of ...

Su, P. F.; Wu, F. Y.

1996-01-01

255

Association between alpha-hemolysin production and HeLa cell-detaching activity in fecal isolates of Escherichia coli.  

OpenAIRE

Escherichia coli isolates that cause detachment of cell monolayers during in vitro adherence assays (cell-detaching E. coli [CDEC]) were recently reported as a potential new group of enteropathogenic bacteria. In the present study, 269 E. coli isolates from feces of children 1 to 5 years of age were identified as CDEC in a detaching assay developed with HeLa cells. The great majority of these isolates were hemolytic within 3 h of growth on blood agar plates and hybridized with a DNA probe for...

Marques, L. R.; Abe, C. M.; Griffin, P. M.; Gomes, T. A.

1995-01-01

256

X-ray-induced inhibition of DNA synthesis in Chinese hamster ovary, human HeLa and mouse L cells  

International Nuclear Information System (INIS)

An X-ray dose of 500 rad inhibits DNA synthesis in Chinese hamster ovary (CHO) cells, in HeLa S3 cells, and in mouse L cells. In no case can the inhibition we observed be explained on the basis of a pool effect or any other process that causes deficient use of exogenously supplied DNA precursors. In all cases the inhibition is mediated by a decrease in replicon initiation processes. Since this inhibition is observed at doses too low to affect individual replicons, we hypothesize that a single hit within a cluster of replicons in some manner blocks the initiation of all replicons within the cluster

257

ToF-SIMS 3D Imaging of Native and Non-Native Species within HeLa Cells  

OpenAIRE

In this study a non-native chemical species, bromodeoxyuridine (BrdU), was imaged within single HeLa cells using time-of-flight secondary ion mass spectrometry (ToF-SIMS). Z-corrected 3D images were reconstructed that accurately portray the distribution of intracellular BrdU as well as other intracellular structures. The BrdU was localized to the nucleus of cells, whereas structures composed of CxHyOz? were located in bundles on the periphery of cells. The CxHyOz? sub-cellular features ha...

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

2013-01-01

258

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

OpenAIRE

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

259

B7-H4 downregulation induces mitochondrial dysfunction and enhances doxorubicin sensitivity via the cAMP/CREB/PGC1-? signaling pathway in HeLa cells.  

Science.gov (United States)

B7-H4 is a B7 family coregulatory protein that inhibits T cell-mediated immunity. B7-H4 is overexpressed in various cancers; however, the functional role of B7-H4 in cancer metabolism is poorly understood. Because mitochondria play pivotal roles in development, proliferation, and death of cancer cells, we investigated molecular and functional alterations of mitochondria in B7-H4-depleted HeLa cells. In a human study, overexpression of B7-H4 was confirmed in the cervices of adenocarcinoma patients (n = 3) compared to noncancer patients (n = 3). In the cell line model, B7-H4 depletion was performed by transfection with small interfering RNA (siRNA). B7-H4 depletion suppressed oxygen consumption rate, ATP production, and mitochondrial membrane potential and mass and increased reactive oxygen species production. In particular, electron transport complex III activity was significantly impaired in siB7-H4-treated cells. Coincidently, depletion of B7-H4 suppressed major mitochondrial regulators (peroxisome proliferator-activated receptor gamma coactivator 1-alpha [PGC1-?] and mitochondrial transcription factor A), a component of oxidative phosphorylation (ubiquinol-cytochrome c reductase core protein 1), and an antiapoptosis protein (Bcl-XL). Mitochondrial dysfunction in siRNA-treated cells significantly augmented oxidative stress, which strongly activated the JNK/P38/caspase axis in the presence of doxorubicin, resulting in increased apoptotic cell death. Investigating the mechanism of B7-H4-mediated mitochondrial modulation, we found that B7-H4 depletion significantly downregulated the cAMP/cAMP response element-binding protein/PGC1-? signaling pathway. Based on these findings, we conclude that B7-H4 has a role in the regulation of mitochondrial function, which is closely related to cancer cell physiology and drug sensitivity. PMID:24658911

Kim, Hyoung Kyu; Song, In-Sung; Lee, Sun Young; Jeong, Seung Hun; Lee, Sung Ryul; Heo, Hye Jin; Thu, Vu Thi; Kim, Nari; Ko, Kyung Soo; Rhee, Byoung Doo; Jeong, Dae Hun; Kim, Young Nam; Han, Jin

2014-12-01

260

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

International Nuclear Information System (INIS)

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

261

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

Energy Technology Data Exchange (ETDEWEB)

HeLa S/sub 3/ 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-(/sup 35/S)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 S/sub 3/ cells migrated slightly faster than the term placental alkaline phosphatase mRNA. Butyrate also induced a second still faster migrating alkaline phosphatase mRNA. Both prednisolone and butyrate increased the steady-state levels of placental alkaline phosphatase mRNA. The data indicate that the increase in phosphatase mRNA by prednisolone and butyrate resulted in the induction of alkaline phosphatase activity and biosynthesis in HeLa S/sub 3/ cells. Furthermore, both agents induced the expression of different alkaline phosphatase gene transcripts without altering its protein product.

Chou, J.Y.; Takahashi, S.

1987-06-16

262

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

Science.gov (United States)

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

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

2014-08-01

263

Cell killing and division delay in asynchronous and synchronized HeLa cells irradiated with alpha particles or x rays  

International Nuclear Information System (INIS)

HeLa cells irradiated with a single or two split doses of ? particles or X rays were observed with time-lapse photography or examined for their colony-forming ability. The cell cycle-dependent variation of cell killing and division delay were compared in synchronous and asynchronous cell populations. Cellular damage by ? particles was manifested in the form of cessation of division, or death, rather than partial division which was predominant for X irradiation. The pattern of cell killing with ? particles was similar to that found with X rays, in that high sensitivity was noted at or close to mitosis, while a resistant peak remained at late S but not in early G1. The pattern of division delay was similar for X rays and ? particles during G2-M, with a maximum delay at mid G2 and no delay past the transition point, but differed during G1-S. During this period, division delay increased with cell age, whereas it showed a broad peak at G1-S boundary and a trough at late S for X rays. However, such was not the case for ? particles

264

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

265

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

Science.gov (United States)

Thymoquinone (TQ) is the active ingredient extracted from the essential oil of Nigella sativa. A number of studies implicated TQ as an antitumor agent. In this study, cytotoxic effects of the oil of N. sativa and TQ were evaluated on human cervical cancer cell line, HeLa cells. IC50 value was ~0.125 ?l/ml for N. sativa oil preparations and 12.5 ?M for TQ. TQ strongly inhibited wound healing at all concentrations ranging from 12.5 to 100 ?M in a scratch wound healing assay. Additionally, induction of apoptosis by TQ was assessed by Giemsa staining and TQ was found to induce apoptosis in cancer cells especially at concentrations of 50 and 100 ?M. TQ-mediated transcriptional regulation of 84 genes involved in apoptosis was studied using a PCR array. At low dose (12.5 ?M), TQ was found to induce expression of four pro-apoptotic genes: BIK (~22.7-fold), FASL (~2.9-fold), BCL2L10 (~2.1-fold), and CASP1 (~2-fold). TQ was also found to reduce the expression of an anti-apoptotic gene implicated in NF-kappa-B signaling and cancer: RELA (~8-fold). At high dose (100 ?M), TQ mediated the expression of 21 genes implicated directly in apoptosis (6 genes), TNF signaling (10 genes), and NF-kappa-B signaling (3 genes) such as BIK, BID, TNFRSF10A, TNFRSF10B, TNF, TRAF3, RELA, and RELB. In conclusion, this study implicates the role of TQ in the inhibition of cancer cell proliferation and migration. At the same time, our results strongly suggest that TQ intervenes with TNF and NF-kappa-B signaling during TQ-mediated induction of apoptosis in cancer cells. PMID:23943306

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

2013-11-01

266

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

International Nuclear Information System (INIS)

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

267

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

International Nuclear Information System (INIS)

The effects of infection with the herpes virus pseudorabies virus on the metabolism of HeLa cell ribosomal RNA were examined. There is a decline both in the synthesis of nucleolar 45S ribosomal precursor RNA and in its processing to mature cytoplasmic RNA. The methylated oligonucleotides in the ribosomal RNA species were studied. The methylation of cytoplasmic ribosomal RNA was essentially unchanged. However there was some undermethylation of the nucleolar precursor. If undermethylated RNA does not mature then this may partly explain the reduced processing in the infected cells. (Author)

268

Oxidation of glutamine in HeLa cells: role and control of truncated TCA cycles in tumour mitochondria.  

Science.gov (United States)

The oxidative metabolism of glutamine in HeLa cells was investigated using intact cells and isolated mitochondria. The concentrations of the cytoplasmic amino acids were found to be aspartate, 8.0 mM; glutamate, 22.2 mM; glutamine, 11.3 mM; glycine, 9.8 mM; taurine, 2.3 mM; and alanine, mitochondria were incubated in a medium containing either glutamine, glutamate, or glutamate plus malate. The transaminase inhibitor AOA inhibited both aspartate efflux from the mitochondria and respiration. The addition of 2-oxoglutarate failed to relieve glutamate plus malate respiration, indicating that 2-oxoglutarate is part of a well-coupled truncated cycle, of which aspartate aminotransferase has been shown to be a component [Parlo and Coleman (1984): J Biol Chem 259:9997-10003]. This was confirmed by the observation that, although it inhibited respiration, AOA did not affect the efflux of citrate from the mitochondria. Thus citrate does not appear to be a cycle component and is directly transported to the medium. Therefore, it was concluded that the truncated TCA cycle in HeLa cells is the result of both a low rate of citrate synthesis and an active citrate transporter. DNP (10 microM) induced a state III-like respiration only in the presence of succinate, which supports the evidence that NAD-linked dehydrogenases were not coupled to respiration, and suggests that these mitochondria may have a defect in complex I of the electron transport chain. Arising from the present results with HeLa cells and results extant in the literature, it has been proposed that a major regulating mechanism for the flux of glutamate carbon in tumour cells is the competitive inhibition exerted by 2-oxoglutarate on aspartate and alanine aminotransferases. This has been discussed and applied to the data. PMID:9443077

Piva, T J; McEvoy-Bowe, E

1998-02-01

269

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

Science.gov (United States)

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

270

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

International Nuclear Information System (INIS)

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.

271

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

Energy Technology Data Exchange (ETDEWEB)

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

272

Increased Activity of Cell Surface Peptidases in HeLa Cells Undergoing UV-Induced Apoptosis Is Not Mediated by Caspase 3  

OpenAIRE

We have previously shown that in HeLa cells treated with a variety of agents there is an increase in cell surface peptidase (CSP) activity in those cells undergoing apoptosis. The increase in CSP activity observed in UVB-irradiated cells undergoing apoptosis was unaffected when the cultures were treated with the aminopeptidase inhibitor bestatin, and matrix metalloprotease inhibitor BB3103, but greatly enhanced when treated with the caspase 3 inhibitor-DEVD, and reduced in the presence of the...

Ellem, Kay A. O.; Winterford, Clay M.; Davern, Catherine M.; Hall, Paula M.; Piva, Terrence J.

2012-01-01

273

Photodynamic effects induced by meso-tris(pentafluorophenyl)corrole and its cyclodextrin conjugates on cytoskeletal components of HeLa cells.  

Science.gov (United States)

The aim of this work was to synthesize new corrole ?-cyclodextrin conjugates ?CD1 (with one ?-cyclodextrin moiety) and ?CD2 (with two ?-cyclodextrin moieties) from 5,10,15-tris(pentafluorophenyl)corrole (TPFC) and to test in vitro the efficacy of these compounds towards tumoral HeLa cells. No dark cytotoxicity was observed for TPFC and ?CD1 at the concentration used for PDT cell treatment, even during long incubation periods (24 h). Fluorescence microscopy showed that TPFC and ?CD1 accumulate in HeLa cells at lysosomes and in the Golgi apparatus, respectively. The cell survival after the PDT treatment with visible light was dependent on light exposure level and compound concentration. ?CD1 was able to penetrate efficiently in the cytoplasm of the HeLa cells. In particular, we have analyzed the photodynamic effect of the corrole derivatives on the microtubules of HeLa cells and the morphological alterations on the mitotic spindle. TPFC and ?CD1 caused photocytotoxicity in tumoral HeLa cells and induced a rapid metaphase blockage of cells that also showed clearly altered configurations of the mitotic spindle. The results showed that TPFC has the highest photosensitizing efficiency on tumoral cells. PMID:25549553

Barata, Joana F B; Zamarrón, Alicia; Neves, M Graça P M S; Faustino, M Amparo F; Tomé, Augusto C; Cavaleiro, José A S; Röder, Beate; Juarranz, Ángeles; Sanz-Rodríguez, Francisco

2015-03-01

274

Confocal Raman imaging for cancer cell classification  

Science.gov (United States)

We propose confocal Raman imaging as a label-free single cell characterization method that can be used as an alternative for conventional cell identification techniques that typically require labels, long incubation times and complex sample preparation. In this study it is investigated whether cancer and blood cells can be distinguished based on their Raman spectra. 2D Raman scans are recorded of 114 single cells, i.e. 60 breast (MCF-7), 5 cervix (HeLa) and 39 prostate (LNCaP) cancer cells and 10 monocytes (from healthy donors). For each cell an average spectrum is calculated and principal component analysis is performed on all average cell spectra. The main features of these principal components indicate that the information for cell identification based on Raman spectra mainly comes from the fatty acid composition in the cell. Based on the second and third principal component, blood cells could be distinguished from cancer cells; and prostate cancer cells could be distinguished from breast and cervix cancer cells. However, it was not possible to distinguish breast and cervix cancer cells. The results obtained in this study, demonstrate the potential of confocal Raman imaging for cell type classification and identification purposes.

Mathieu, Evelien; Van Dorpe, Pol; Stakenborg, Tim; Liu, Chengxun; Lagae, Liesbet

2014-05-01

275

Relationship of ROS and NO in X-ray induced bystander effects in HeLa cells  

International Nuclear Information System (INIS)

Accumulating evidence indicates that irradiated cells can release signals which induce a series of biological responses in non-exposed cells. This is known as irradiation-induced bystander effects. Both reactive oxygen species(ROS)and nitric oxide(NO) play important roles in bystander effects. In this study, we determined the relationship of ROS and NO in the signaling pathway of bystander effects. HeLa cells were treated with or without dimethy sulfoxide(DMSO) before X-ray irradiation, and micronuclei formation as well as cell proliferation rate was detected in both irradiated and bystander cells. In addition,we also detected inducible nitric oxide synthase(iNOS)expression and NO level in irradiated cells using Western blotting and DAF-FM DA fluorescent probe, respectively. Our results showed that micronuclei were induced in irradiated and bystander cells while DMSO treatment significantly suppressed the formation of micronuclei in both of them. We also found that when cells were irradiated their proliferation rate was suppressed while DMSO treatment eliminated this inhibition effect.In contrast, the cells received conditioned medium from irradiated cells proliferated more quickly than the cells received medium from non-irradiated cells while DMSO treatment reduced the difference. Finally, we found that irradiated cells had higher level of iNOS and NO compared to non-irradiated controls, whereas DMSO treatment decreased their levels. These results suggest that ROS is levels. These results suggest that ROS is the upstream signal of NO in X-ray induced bystander effects in HeLa cells. (authors)

276

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

Science.gov (United States)

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

277

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

International Nuclear Information System (INIS)

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

278

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in english A sulfated fucan from Laminaria abyssalis marine alga prevented the interaction of HTLV-1 particles, purified from the MT-2 cell line, with HeLa cells. The infection obtained using a concentrated virus suspension was detected only by amplification of the newly synthesized HTLV-1 proviral cDNA by the [...] nested-polymerase chain reaction (PCR). The sulfated polysaccharide was not toxic to the cells at a concentration of 100 µg/mL and prevented infection by the viral particles when added to the cell monolayers. The proviral cDNA was only detected when the sulfated polysaccharide was added to the cells three hours post-infection, indicating that the inhibitory activity occurred in the initial stages of virus-cell interaction. Our results demonstrate, for the first time, the ability of a sulfated fucan from marine algae to inhibit virus transmission through free virus particles.

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

2011-04-01

279

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

280

In AtT20 and HeLa cells brefeldin A induces the fusion of tubular endosomes and changes their distribution and some of their endocytic properties  

OpenAIRE

We have studied the effects of brefeldin A (BFA) on the tubular endosomes in AtT20 and HeLa cells (Tooze, J., and M. Hollinshead. 1991. J. Cell Biol. 115:635-653) by electron microscopy of cells labeled with three endocytic tracers, HRP, BSA-gold, and transferrin conjugated to HRP, and by immunofluorescence microscopy. For the latter we used antibodies specific for transferrin receptor, and, in the case of AtT20 cells, also antibodies specific for synaptophysin. In HeLa cells BFA at concentra...

1992-01-01

281

Extended electrical model for impedance characterization of cultured HeLa cells in non-confluent state using ECIS electrodes.  

Science.gov (United States)

Electric cell substrate impedance sensing has been widely used as a label free quantitative platform to study various cell biological processes and it is extremely essential to extract the parameters like the variation of the cell substrate spacing, changing projected area of the cell on the electrode and approximate cluster size during the non-confluent state to understand the mechanism of proliferation of the cells. The distributed analytical models developed so far to extract these parameters are applicable only under the conditions when the cells have become confluent. There are some lumped electrical models which have been reported for the non-confluent state but they do not provide correct estimate of the changing cell substrate spacing and the cell cluster size during growth. In this paper we develop extended distributed electrical models to characterize the impedance spectroscopy behavior of cultured HeLa cells in 200 Hz to 1 MHz range using eight well ECIS electrodes in the non-confluent state. The distributed model introduces some pseudo regularity in the arrangement of the non-confluent cells to extract the average ensemble of the significant parameters. The parameters extracted from the distributed model after 10 hours, 20 hours, and 30 hours of HeLa cell growth have been compared with the lumped circuit model and has been observed to fit the experimental data with a seven times improved fit quality factor. Further, the changing cell radius and cluster radius extracted at three different instants of time from the distributed analytical model have been found to match closely the microscopic observation in contrast to the lumped circuit model. PMID:23995584

Mondal, D; RoyChaudhuri, C

2013-09-01

282

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

Energy Technology Data Exchange (ETDEWEB)

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

Liu, Xianguang; An, Zhengzhe; Song, Hye Jin; Kim, Won Dong; Park, Woo Yoon [Chungbuk National University College of Medicine, Cheongju (Korea, Republic of); Jang, Seong Soon [The Catholic University of Korea College of Medicine, Seoul (Korea, Republic of); Yu, Jae Ran [Konkuk University College of Medicine, Chungju (Korea, Republic of)

2012-06-15

283

Involvement of protein kinase C in the control of tRNA modification with queuine in HeLa cells.  

OpenAIRE

The eukaryotic tRNA:guanine transglycosylase (TGT) catalyses the base-for-base exchange of guanine for queuine (the q-base)--a nutrition factor for eukaryotes--at position 34 of the anticodon of tRNAsGUN (where 'N' represents one of the four canonical tRNA nucleosides), yielding the modified tRNA nucleoside queuosine (Q). This unique tRNA modification process was investigated in HeLa cells grown under either aerobic (21% O2) or hypoxic conditions (7% O2) after addition of chemically synthesiz...

Langgut, W.; Reisser, T.

1995-01-01

284

Properties of the deoxycholate-solubilized HeLa cell plasma membrane receptor for binding group B coxsackieviruses.  

OpenAIRE

Physical and chemical properties of deoxycholate-solubilized HeLa cell plasma membrane receptors for binding group B coxsackieviruses were determined. Receptors eluted from Sepharose 4B with an apparent molecular weight of 275,000 and sedimented with an S value of between 14.7 and 4.9 and a buoyant density of 1.06 to 1.10 g/cm3. Virus-binding activity was destroyed after treatment with proteases, glycosidases, and periodate but was unaffected by lipases or reducing or alkylating agents. Addit...

Krah, D. L.; Crowell, R. L.

1985-01-01

285

Uracil DNa-glycosylase from HeLa cells: general properties, substrate specificity and effect of uracil analogs.  

OpenAIRE

Uracil-DNA glycosylase was partially purified from HeLa cells. Various substrates containing [3H]dUMP residues were prepared by nick-translation of calf thymus DNA. The standard substrate was double-stranded DNA with [3H]dUMP located internally in the chain. Compared to the release of uracil from this substrate, a 3-fold increase in the rate was seen with single-stranded DNA, and a 20-fold reduction in the rate was observed when the [3H]dUMP-residue was located at the 3'end. The rate of [3H]u...

Krokan, H.; Wittwer, C. U.

1981-01-01

286

Mapping of nascent light and heavy strand transcripts on the physical map of HeLa cell mitochondrial DNA.  

OpenAIRE

The sequences complementary to the nascent RNA molecules isolated from transcription complexes of HeLa cell mtDNA have been mapped on the H and L strands of mtDNA by the S1 protection technique. The distribution of these sequences among different Hpa II restriction fragments was found to reflect the position of these fragments in the Hpa II map of mtDNA. Thus, the S1-resistant hybrids formed with the L strand corresponded almost exclusively to the right half of the genome past the origin of r...

Cantatore, P.; Attardi, G.

1980-01-01

287

[Response of HeLa cells to mitomycine C. I. Cell division].  

Science.gov (United States)

Using light microscopy, time-lapse imaging, and digital image analysis, the effect of mitomycine C (10 ?g/ml) on HeLa-M cells has been studied. It has been shown that, after a 2 h contact with mitomycine, the cells could be separated into 2 groups: M-1--the functional cells surviving after division but non-entering mitosis any more; M-II--the cells entering mitosis but incapable to finish it; they are lost. Mitomycine C is known to specifically block DNA replication being located in the DNA minor groove. It should inhibit PHK synthesis if one follows the standard hypothesis of a transcription bubble formation. However, increasing the cell and nucleolus area during the M-I cell growth suggests that RNA and protein synthesis is not blocked. The author concludes that the presented data confirm his hypothesis about RNA synthesis in the major DNA groove (Petrov, 2006). PMID:25508685

2013-01-01

288

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

289

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

OpenAIRE

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

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

2013-01-01

290

[Response of HeLa cells to mitomycine C. II. Morphometry of the cells].  

Science.gov (United States)

HeLa-M cells were analyzed after the 2h incubation in the medium with mitomycin C (10 ?g/ml). It has been shown that a part of the cells contacted with the cytostatic agent passes mitosis normally, but the daughter cells no longer divide. During the observation period (2 days), the area of the cells increased linearly reaching twice the size of intact cells. Thereby spreading of the cells contacted with mitomycine decreased, and their polarisation was not changed. Along with the increasing cell size, the nucleus size is also increased indicated de novo protein synthesis. Analysis of nucleoli revealed a small decrease of their area during the first hours after the contact with mitomycine followed by an increase of the area. Finally, the nucleolus area exceeded that of control cells, which can only happen if rRNA was synthesized. On the other hand, DNA cross-linking by mitomycine should inhibit transcription because the mechanism of transcription is assumed to involve local melting of DNA within its minor groove. This contradiction can be overcome by assuming that transcription occurs in a DNA major groove without separation of the chains. PMID:25508686

2013-01-01

291

Crinane alkaloids of the amaryllidaceae with cytotoxic effects in human cervical adenocarcinoma (HeLa) cells.  

Science.gov (United States)

The family Amaryllidaceae has a long history of usage in the traditional medicinal practices of the indigenous peoples of South Africa, with three of its species known to be used for cancer treatment. Furthermore, the Amaryllidaceae is widely recognized for its unique alkaloid constituents, several of which exhibit potent and selective cytotoxic activities. In this study, several crinane alkaloids derived from local Amaryllidaceae species were examined for cytotoxic effects against the human cervical adenocarcinoma cell line, of which distichamine was the most potent (IC50 2.2 microM). PMID:24868855

Nair, Jerald J; Rárová, Lucie; Strnad, Miroslav; Bastida, Jaume; Cheesman, Lee; van Staden, Johannes

2014-04-01

292

Possible attenuation of the G2 DNA damage cell cycle checkpoint in HeLa cells by extremely low frequency (ELF) electromagnetic fields  

OpenAIRE

Abstract Background The issue remains unresolved as to whether low frequency magnetic fields can affect cell behaviour, with the possibility that they may be in part responsible for the increased incidence of leukaemia in parts of the population exposed to them. Methods Combined treatment of HeLa cells with gamma-irradiation (1, 3 and 5 Grays) and extra low frequency magnetic fields of ~50 Hz was carried out under rigorously controlled conditions. Results...

Heaton Brian; Lamb Justin; Harris Paul A; Wheatley Denys N.

2002-01-01

293

The influence of digoxin antibodies on digoxin disposition and effect: studies in guinea-pigs and HeLa cells.  

Science.gov (United States)

Pretreatment of guinea-pigs with digoxin-specific Fab (fragment antigen binding) fragments reduced the cardiotoxicity of intravenously infused digoxin (the lethal doses in Fab-treated and control animals were 1.0 and 0.6 mgkg-1, respectively). At death the serum digoxin concentration was elevated 2 fold in the Fab-treated animals, while the tissue concentrations were generally lower. The 30-40% lower cardiac digoxin concentration (seen in whole homogenate and throughout the subcellular fractions examined) was surprising; presumably this reflects a difference from the controls in the proportion of pharmacologically active/inactive digoxin in this organ. Adding digoxin-specific immunoglobulin G or the Fab fragments to HeLa cells before incubation with digoxin, reduced specific digoxin binding (Na pump-bound) slightly more than the non-specific binding. Adding specific antibody after digoxin, however, did not reduce digoxin binding or effect a recovery in Na pump activity. It seems that the protective effect of digoxin-specific antibodies seen in the guinea-pig can to some extent be simulated using HeLa cells. However, this is apparently not so regarding the widely-reported ability of these antibodies to reverse the action of digoxin. PMID:3978308

Griffiths, N. M.; Hewick, D. S.; Lamb, J. F.; Stevenson, I. H.

1985-01-01

294

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

International Nuclear Information System (INIS)

An in vitro poliovirus RNA-synthesizing system derived from a crude membrance fraction of infected HeLa cells was used to analyze the mechanism of initiation of poliovirus plus-strand RNA synthesis. This system contains an activity that synthesizes the nucleotidyl proteins VPg-pU and VPg-pUpU. These molecules represent the 5'-terminal structure of nascent RNA molecules and of virion RNA. The membranous replication complex is also capable of synthesizing mucleotidyl proteins containing nine or more of the poliovirus 5'-proximal nucleotides as assayed by the formation of the RNase 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

295

The PPAR? agonist L-165041 promotes VEGF mRNA stabilization in HPV18-harboring HeLa cells through a receptor-independent mechanism.  

Science.gov (United States)

Peroxisome Proliferator-Activated Receptor-? (PPAR?) is a ligand-inducible transcription factor activated by both natural (fatty acids and derivatives) and high affinity synthetic agonists. It is thought to play a role in angiogenesis development and Vascular Endothelial Growth Factor (VEGF) regulation but its contribution remains unclear. Until now, the PPAR? agonism effect on VEGF expression in cervical cancer cells was unknown. This led to our interest in assessing the effect of PPAR? activation on the regulation of different VEGF isoforms mRNA expression and the impact of E6 viral oncoprotein and its target p53 on this regulation in cervical cancer cells. Here, we showed that the PPAR? agonist L-165041 induces VEGF(121), VEGF(165) and VEGF(189) expression in HPV (Human Papillomavirus) positive HeLa cells but not in HPV negative cells. The underlying mechanisms did involve neither E6 oncoprotein nor p53. We highlighted a novel mode of PPAR? ligand action including a post-transcriptional regulation of VEGF mRNA expression through the p38 MAPK signaling pathway and the activation of the mRNA-stabilizing factor HuR. But most importantly, we clearly demonstrated that L-165041 acts independently of PPAR? since its effect was not reversed by a chemical inhibition with a specific antagonist and the siRNA-mediated knockdown of the nuclear receptor. As VEGF is crucial for cancer development, the impact of PPAR? ligands on VEGF production is of high importance. Thus, the molecular mechanism of their action has to be elucidated and as a result, PPAR? agonists currently in clinical trials should be carefully monitored. PMID:24172859

Roche, Emmanuelle; Lascombe, Isabelle; Bittard, Hugues; Mougin, Christiane; Fauconnet, Sylvie

2014-02-01

296

Pitx2a Expression Alters Actin-Myosin Cytoskeleton and Migration of HeLa Cells through Rho GTPase SignalingV?  

OpenAIRE

We ectopically expressed the transcription factor Pitx2a, one of the Pitx2 isoforms, in HeLa cells by using a tetracycline-inducible expression system and examined whether Pitx2a was capable of modulating Rho GTPase signaling and altering the cell's cytoskeleton. Ectopic expression of Pitx2a induced actin-myosin reorganization, leading to increased cell spreading, suppression of cell migration, and the strengthening of cell-cell adhesion, marked by the accumulation and localization of ?-cate...

Wei, Qize; Adelstein, Robert S.

2002-01-01

297

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

OpenAIRE

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....

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

2014-01-01

298

Gene expression responses of HeLa cells to chemical species generated by an atmospheric plasma flow.  

Science.gov (United States)

Plasma irradiation generates many factors able to affect the cellular condition, and this feature has been studied for its application in the field of medicine. We previously reported that hydrogen peroxide (H2O2) was the major cause of HeLa cell death among the chemical species generated by high level irradiation of a culture medium by atmospheric plasma. To assess the effect of plasma-induced factors on the response of live cells, HeLa cells were exposed to a medium irradiated by a non-lethal plasma flow level, and their gene expression was broadly analyzed by DNA microarray in comparison with that in a corresponding concentration of 51 ?M H2O2. As a result, though the cell viability was sufficiently maintained at more than 90% in both cases, the plasma-medium had a greater impact on it than the H2O2-medium. Hierarchical clustering analysis revealed fundamentally different cellular responses between these two media. A larger population of genes was upregulated in the plasma-medium, whereas genes were downregulated in the H2O2-medium. However, a part of the genes that showed prominent differential expression was shared by them, including an immediate early gene ID2. In gene ontology analysis of upregulated genes, the plasma-medium showed more diverse ontologies than the H2O2-medium, whereas ontologies such as "response to stimulus" were common, and several genes corresponded to "response to reactive oxygen species." Genes of AP-1 proteins, e.g., JUN and FOS, were detected and notably elevated in the plasma-medium. These results showed that the medium irradiated with a non-lethal level of plasma flow altered various gene expressions of HeLa cells by giving not only common effects with H2O2 but also some distinctive actions. This study suggests that in addition to H2O2, other chemical species able to affect the cellular responses exist in the plasma-irradiated medium and provide unique features for it, probably increasing the oxidative stress level. PMID:24996177

Yokoyama, Mayo; Johkura, Kohei; Sato, Takehiko

2014-08-01

299

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

OpenAIRE

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 IC5...

Villar, Jose? A. F. P.; Viana, Gustavo H. R.; Barbosa, Leandro A.; Varotti, Fernando P.; Simo?es, Sarah C.; Andersson Barison; Souza, Estrela M. P. V. E.; Da Silva, Marina G.; Da Silva, Graziele D.

2012-01-01

300

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

301

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

International Nuclear Information System (INIS)

The effects of uv irradiation on the incorporation of [3H]uridine in HeLa (human) cell mRNA, rRNA, heterogeneous nuclear RNA (hnRNA) and early mRNA from adenovirus type 2 have been compared. The uv target size of cell mRNA is at least 3 times larger than the average size of the mRNA itself and larger than the adenovirus-2 early mRNA, which is known to derive from transcription units of about 1.5-5.0 kilobases. The uv target size of hnRNA, in contrast, is about the same as its size determined by sedimentation and overlaps with the target size of mRNA. It is concluded that most mRNA derives from a higher molecular weight hnRNA molecule

302

Molecular mechanism of inhibitory effects of C-phycocyanin combined with all-trans-retinoic acid on the growth of HeLa cells in vitro.  

Science.gov (United States)

We studied the effects of all-trans-retinoic acid (ATRA), C-phycocyanin (C-PC), or ATRA+C-PC on the growth of cervical cells (HeLa cells), cell cycle distribution, and apoptosis. The anticancer mechanism of the drug combination was revealed. MTT assay was adopted to determine the effects of C-PC and ATRA on the growth of HeLa cells. The expression quantities of cyclin-dependent kinase (CDK) 4, cyclin D1, Bcl-2, caspase-3, and CD59 were determined by in situ hybridization, immunofluorescence, immunohistochemistry staining, Western blot, and RT-PCR. TUNEL assay was adopted to determine the cellular apoptosis levels. Both C-PC and ATRA could inhibit the growth of HeLa cells, and the combination of ATRA+C-PC functioned cooperatively to induce apoptosis in HeLa cells. The dosage of ATRA was reduced when it cooperated with C-PC to reduce the toxicity. ATRA treated with C-PC could induce more cell cycle arrests than the single drug used by decrease in cyclin D1 and CDK4 expression. The combination of the two drugs could upregulate caspase-3 and downregulate the Bcl-2 gene and induce cell apoptosis. Moreover, the combination therapy has an important immunological significance in decreased expression of the CD59 protein. Singly, C-PC or ATRA could inhibit the growth of HeLa cells, and the effects of treatment were further enhanced in the combination group. In combination with C-PC, the dosage of ATRA was effectively reduced. The C-PC?+?ATRA combination might take effect by inhibiting the progress of the cell cycle, inducing cell apoptosis and promoting complement-mediated cytolysis. PMID:24563337

Yang, Fan; Li, Bing; Chu, Xian-Ming; Lv, Cong-Yi; Xu, Ying-Jie; Yang, Peng

2014-06-01

303

Secretion of apolipoprotein B-containing lipoproteins from HeLa cells is dependent on expression of the microsomal triglyceride transfer protein and is regulated by lipid availability.  

Science.gov (United States)

To elucidate the role of the microsomal triglyceride transfer protein (MTP) in lipoprotein assembly, MTP and apolipoprotein B-53 (apoB 53; the N-terminal 53% of apoB) were expressed in HeLa cells. The results showed that apoB-53 could be expressed in HeLa cells with or without expression of MTP. In contrast, efficient secretion of apoB-53 required expression of MTP. Ultracentrifugal density flotation analysis showed that apoB-53 was secreted predominantly as a particle with the density of high density lipoprotein. An essentially identical apoB-53 particle density distribution was obtained after transient expression of apoB-53 in McArdle RH-7777 rat hepatoma cells. The mass of apoB-53 secreted was greater, and the flotation density was lower, from cells fed lipid, suggesting that apoB secretion in HeLa cells was regulated by lipid availability, similar to what has been described for lipoprotein-producing cell lines. These results indicate that MTP is necessary and sufficient to direct the regulated secretion of apoB-53 in HeLa cells. Images PMID:8052632

Gordon, D A; Jamil, H; Sharp, D; Mullaney, D; Yao, Z; Gregg, R E; Wetterau, J

1994-01-01

304

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

International Nuclear Information System (INIS)

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

305

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

Energy Technology Data Exchange (ETDEWEB)

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

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

2013-09-06

306

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

International Nuclear Information System (INIS)

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 raells 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

307

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

International Nuclear Information System (INIS)

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

308

Inhibition of the MRP1-mediated transport of the menadione-glutathione conjugate (thiodione) in HeLa cells as studied by SECM  

OpenAIRE

Oxidative stress induced in live HeLa cells by menadione (2-methyl-1,4-napthaquinone) was studied in real time by scanning electrochemical microscopy (SECM). The hydrophobic molecule menadione diffuses through a living cell membrane where it is toxic to the cell. However, in the cell it is conjugated with glutathione to form thiodione. Thiodione is then recognized and transported across the cell membrane via the ATP-driven MRP1 pump. In the extracellular environment, thiodione was detected by...

Koley, Dipankar; Bard, Allen J.

2012-01-01

309

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

310

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

311

Loss of FADS2 Function Severely Impairs the Use of HeLa Cells as an In Vitro Model for Host Response Studies Involving Fatty Acid Effects  

Science.gov (United States)

Scope Established epithelial cell lines equipped with pattern recognition receptors such as the Toll-like receptor (TLR)-2 are common tools for immune response studies on invading pathogens, e.g. the obligate intracellular species of Chlamydia. Moreover, such models are widely used to elucidate fatty acid-mediated immune effects. In several transformed cell lines, however, unusual loss of metabolic functions was described. The cell lines A549 and HeLa are poorly characterized in this respect. Therefore, we comparatively assessed the metabolic capacity of A549 and HeLa prior to proposed application as in vitro model for fatty acid effects on chlamydial infection. Methodology/Principal Findings We incubated both cell lines either with substrates (C18?2n?6 or C18?3n?3) or products (C18?3n?6, C18?4n?3) of fatty acid desaturase-2 (FADS2), and analysed the fatty acid profiles after 24 h and 72 h by gas chromatography. Based on these data, we suspected that the complete discontinuation of normal biosynthesis of long-chain polyunsaturated fatty acids (LC-PUFA) in HeLa was due to loss of FADS2 function. Consequently, prostaglandin E2 (PGE2) formation was less inducible by TLR2 stimulation in HeLa, likely as a result of not only insufficient supply of precursors but also weak cyclooxygenase-2 (COX-2) response. In accordance, Chlamydia infection rates were consistently lower in HeLa than in A549. Sequence analysis revealed no alteration within the FADS2 gene in HeLa. The FADS2 expression level, however, was significantly lower and, in contrast to A549, not regulated by C18?2n?6. A549 exhibited regular fatty acid metabolism and enzyme functionality. Conclusions/Significance Our data show that HeLa cells considerably differ from A549 at several stages of fatty acid metabolism. The poor metabolic potential of HeLa, mainly concerning FADS2 upstream of COX-2 function, calls into question whether these cells represent a good model to unveil fatty acid or downstream eicosanoid effects in the course of intracellular bacterial infection. PMID:25549244

Jaudszus, Anke; Degen, Christian; Barth, Stephan W.; Klempt, Martin; Schlörmann, Wiebke; Roth, Alexander; Rohrer, Carsten; Sauerwein, Helga; Sachse, Konrad; Jahreis, Gerhard

2014-01-01

312

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

313

Human Immunodeficiency Virus Type 1 Attachment to HeLa CD4 Cells Is CD4 Independent and gp120 Dependent and Requires Cell Surface Heparans  

OpenAIRE

The binding of human immunodeficiency virus type 1 (HIV-1) (Hx10) virions to two different cell lines was analyzed by using a novel assay based on the detection, by anti-HLA-DR-specific antibodies, of HLA-DR+ virus binding to HLA-DR- cells. Virion attachment to the CD4+-T-cell line A3.01 was highly CD4 dependent in that it was potently inhibited by CD4 monoclonal antibodies (MAbs), and little virus binding to the CD4- sister A2.01 line was observed. By contrast, virion binding to HeLa cells e...

Mondor, I.; Ugolini, S.; Sattentau, Qj

1998-01-01

314

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

International Nuclear Information System (INIS)

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

315

Fluorescence studies on the role of tryptophan in heterogeneous nuclear ribonucleoprotein particles of HeLa cells.  

Science.gov (United States)

The 40 S heterogeneous nuclear ribonucleoprotein (hnRNP) particles from HeLa cells reveal tryptophan fluorescence with a bi-exponential decay, indicating that only a few of the 'core' proteins contain tryptophan residues. The presence of tryptophan residues distinguishes hnRNP particles from nucleosomes, with which they otherwise share a number of properties. This difference, however, is not essential for protein-RNA binding, as the fluorescence decay remains unchanged when hnRNP particles are dissociated into protein and RNA. However, the Stern-Volmer quenching constant is doubled upon salt dissociation, i.e. tryptophan residues become more accessible to solvent. Thus tryptophan quenching is a useful parameter for monitoring protein-protein interactions in hnRNP particles. PMID:2604698

Schenkel, J; Appel, I; Schwarzwald, R; Bautz, E k; Wolfrum, J; Greulich, K O

1989-01-01

316

Analysis of the factors in determining radiosensitivity in mammalian cells by using radio-sensitive and -resistant clones isolated from HeLa S3 cells in vitro  

International Nuclear Information System (INIS)

The factors in determining radiosensitivity of cultured mammalian cells were analysed by using two clones each having different radiosensitivities. The radiosensitive clones were isolated from HeLa S3 cells by the N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)-treatment, X-irradiation (200 R) and 5-bromodeoxyuridine (BUdR)-visible light method. On the other hand, the radioresistant clone was isolated by single X-irradiation (2000 R) from MNNG-treated HeLa S3 cell population. The radiosensitivities expressed in D sub(o) and D sub(q) values were 110 and 140 R in radiosensitive SM-1a clone and 180 and 230 R in radioresistant RM-1b clone respectively. The biological and biochemical characteristics of both clones such as the distribution of chromosome numbers, formation and rejoining of single strand breaks in DNA caused by X-irradiation, non-protein sulfhydryl (NPSH) and apparent total sulfhydryl (APSH) contents were measured. Among the characteristics analysed, different contents of NPSH in the cell were well correlated to their daiosensitivities among the original HeLa S3 cells, SM-1a and RM-1b clone. Additionally, it was found that the radioresistant L.P3 Co-3 cells isolated by Tsuboi et al. from the original mouse L.P3 cells by means of serial irradiation with 60Co ?-rays have more abundant NPSH than the original L.P3 cells. From these results, it can be concluded that the amount of NPSH play the main role in determining radiosensitivity in cultured mammalianng radiosensitivity in cultured mammalian cells. (auth.)

317

Thermotolerance induced at a mild temperature of 40°C alleviates heat shock-induced ER stress and apoptosis in HeLa cells.  

Science.gov (United States)

Hyperthermia (39-45°C) has emerged as an alternate prospect for cancer therapy in combination with radiation and chemotherapy. Despite promising progress in the clinic, molecular mechanisms involved in hyperthermia-induced cell death are not clear. Hyperthermia causes protein denaturation/aggregation, which results in cell death by apoptosis and/or necrosis. Hyperthermia also induces thermotolerance, which renders cells resistant to subsequent exposure to lethal heat shock. This study investigates the role of both lethal (42-43°C) and mild (40°C) hyperthermia in regulating ER stress and ER stress-induced apoptosis in HeLa cells. The ability of mild thermotolerance induced at 40°C to alleviate either or both of these processes is also determined. Hyperthermia (42-43°C) induced ER stress, revealed by phosphorylation of PERK, eIF2? and IRE1?, cleavage of ATF6 and increased expression of BiP and sXBP1. Real-time PCR revealed that mRNA levels of ATF6, ATF4, BiP, sXBP1 and CHOP increased in cells exposed to hyperthermia. Moreover, hyperthermia caused disruption of calcium homeostasis and activated the calpain-calpastatin proteolytic system and ER resident caspase 4. Pre-exposure to mild hyperthermia (40°C) alleviated the induction of cytotoxicity and ER stress by hyperthermia (42-43°C) and protected cells against ER stress-induced apoptosis. ShRNA-mediated depletion of Hsp72 abrogated protective effects of mild thermotolerance (40°C) against heat-shock induced ER stress and sensitized cells to ER stress-mediated apoptosis. Our findings show that Hsp72 contributes to the protective effects of mild hyperthermia (40°C) against hyperthermia-induced ER stress and apoptosis. PMID:25260982

Bettaieb, Ahmed; Averill-Bates, Diana A

2015-01-01

318

In vivo synthesis of selenium nanoparticles by Halococcus salifodinae BK18 and their anti-proliferative properties against HeLa cell line.  

Science.gov (United States)

Nanoparticles synthesis by bacteria and yeasts has been widely reported, however, synthesis using halophilic archaea is still in a nascent stage. This study aimed at the intracellular synthesis of selenium nanoparticles (SeNPs) by the haloarchaeon Halococcus salifodinae BK18 when grown in the presence of sodium selenite. Crystallographic characterization of SeNPs by X-ray diffraction, Selected area electron diffraction, and transmission electron microscopy exhibited rod shaped nanoparticles with hexagonal crystal lattice, a crystallite domain size of 28 nm and an aspect ratio (length:diameter) of 13:1. Energy disruptive analysis of X-ray analysis confirmed the presence of selenium in the nano-preparation. The nitrate reductase enzyme assay and the inhibitor studies indicated the involvement of NADH-dependent nitrate reductase in SeNPs synthesis and metal tolerance. The SeNPs exhibited good anti-proliferative properties against HeLa cell lines while being non-cytotoxic to normal cell line model HaCat, suggesting the use of these SeNPs as cancer chemotherapeutic agent. This is the first study on selenium nanoparticles synthesis by haloarchaea. PMID:25219897

Srivastava, Pallavee; Braganca, Judith M; Kowshik, Meenal

2014-01-01

319

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.

320

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

321

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

322

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

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

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

2013-09-01

323

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

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

324

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

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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 traundamental change that develops during transformation

325

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

Scientific Electronic Library Online (English)

Full Text Available SciELO Brazil | Language: English Abstract in portuguese Características de virulência de 56 amostras de Escherichia coli isoladas de eqüinos doentes (secreção de colo uterino, fragmentos de necrópsia do trato gastrointestinal e de pulmões, fezes diarréicas e lavado traqueal) foram examinadas para determinar o padrão de aderência em células HeLa e pesquis [...] ar a presença de genes de virulência de vários patotipos de E. coli. Duas amostras não aderentes apresentaram astA, gene que codifica a toxina termo-estável de E. coli enteroagregativa. Das vinte e sete amostras (48,2%) que aderiram a células HeLa, 21 (77,8%) apresentaram o padrão de aderência agregativa (AA) que caracteriza o patotipo de E. coli Enteroagregativa (EAEC). Nove destas amostras que apresentaram AA foram isoladas de secreção de colo uterino, incluindo uma que apresentava genes de virulência de patotipos de EAEC (aggR,aap,irp2 e pic). Esta é a primeira descrição do fenótipo AA em amostras de cavalos doentes. Estas amostras deverão ser melhor avaliadas em relação a sua potencial função na patogênese de diferentes doenças eqüinas, bem como à possibilidade destes animais representarem um reservatório de infecções humanas causadas por esta bactéria. Abstract in english The virulence attributes of 56 Escherichia coli strains isolated from sick horses (secretions of uterine cervices; gastrointestinal and lung fragments of necropsy; diarrheic feces, and tracheal washings) was examined by determining their adherence pattern to HeLa cells and searching for the presence [...] of virulence genes of the various E. coli pathotypes. Two non-adherent strains presented astA, which encodes the enteroaggregative E. coli heat-stable toxin. Twenty-seven strains (48.2%) adhered to HeLa cells, 21 (77.8%) of which presented the aggregative adherence pattern (AA) that characterize the Enteroaggregative E. coli pathotype (EAEC). Nine of the strains presenting AA were isolated from secretions of uterine cervix, including one carrying virulence genes of the EAEC pathotype (aggR,aap,irp2, and pic). This is the first description of the AA phenotype amongst E. coli strains from sick horses. Such strains should be further evaluated regarding their potential role in the pathogenesis of diverse equine diseases and as reservoirs of human infections.

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

2007-03-01

326

A systems biology analysis of the changes in gene expression via silencing of HPV-18 E1 expression in HeLa cells.  

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Previous studies have reported the detection of a truncated E1 mRNA generated from HPV-18 in HeLa cells. Although it is unclear whether a truncated E1 protein could function as a replicative helicase for viral replication, it would still retain binding sites for potential interactions with different host cell proteins. Furthermore, in this study, we found evidence in support of expression of full-length HPV-18 E1 mRNA in HeLa cells. To determine whether interactions between E1 and cellular proteins play an important role in cellular processes other than viral replication, genome-wide expression profiles of HPV-18 positive HeLa cells were compared before and after the siRNA knockdown of E1 expression. Differential expression and gene set enrichment analysis uncovered four functionally related sets of genes implicated in host defence mechanisms against viral infection. These included the toll-like receptor, interferon and apoptosis pathways, along with the antiviral interferon-stimulated gene set. In addition, we found that the transcriptional coactivator E1A-binding protein p300 (EP300) was downregulated, which is interesting given that EP300 is thought to be required for the transcription of HPV-18 genes in HeLa cells. The observed changes in gene expression produced via the silencing of HPV-18 E1 expression in HeLa cells indicate that in addition to its well-known role in viral replication, the E1 protein may also play an important role in mitigating the host's ability to defend against viral infection. PMID:25297386

Castillo, Andres; Wang, Lu; Koriyama, Chihaya; Eizuru, Yoshito; Jordan, King; Akiba, Suminori

2014-10-01

327

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

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

Nazli C

2012-04-01

328

Antioxidant activity and cytotoxic effect of aviprin and aviprin-3?-O-D-glucopyranoside on LNCaP and HeLa cell lines.  

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Many researchers have shown that plant-derived polyphenolic compounds are helpful nutraceuticals in restraining various disorders such as neoplastic diseases. In this study two linear furanocoumarins, aviprin and aviprin-3?-O-D-glucopyranoside (A3G), were isolated from methanol extract of Prangos uloptera roots. The evaluation of free radical scavenging capacity of the compounds showed that aviprin is a more effective antioxidant than A3G with RC50 of 0.54?mg?mL?¹. The biological and antiproliferative activities of the furanocoumarins were examined using human cervical carcinoma HeLa cell line and LNCaP prostatic cell line. Cell membrane integrity and cell viability were evaluated by measuring trypan blue exclusion assay and reduction of the tetrazolium blue compound, respectively. Treating the LNCaP cell line with various concentrations of the furanocoumarins showed that IC?? of aviprin and A3G were 0.4 and 6.6?mg?mL?¹, whereas their CC50 values were 0.7 and 11?mg?mL?¹, respectively. These results indicated that 42.7% of LNCaP cells were not dead by necrosis. Treating the HeLa cells by the furanocoumarins showed the greater sensitivity of the HeLa cell line than the LNCaP cell line. A morphological analysis and the study of DNA fragmentation provided further some evidence for the inhibition of the LNCaP cell line via apoptosis induction. PMID:21714729

Zahri, Saber; Razavi, Seyed Mehdi; Moatamed, Zahra

2012-01-01

329

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

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

330

Intercellular transfer of a glycosylphosphatidylinositol (GPI)-linked protein: release and uptake of CD4-GPI from recombinant adeno-associated virus-transduced HeLa cells.  

OpenAIRE

A diverse group of GPI-anchored protein structures are ubiquitously expressed on the external cell membranes of eukaryotes. Whereas the physiological role for these structures is usually defined by their protein component, the precise biological significance of the glycolipid anchors remains vague. In the course of producing a HeLa cell line (JM88) that contained a recombinant adeno-associated virus genome expressing a GPI-anchored CD4-GPI fusion protein on the surface of the cells, we noted ...

Anderson, S. M.; Yu, G.; Giattina, M.; Miller, J. L.

1996-01-01

331

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

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

332

Action of caffeine on x-irradiated HeLa cells. I. Delayed inhibition of DNA synthesis  

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Treatment of HeLa S3 cells with 1 mM caffeine delays progression through G1 by 1.5 hours but causes no other detectable inhibition of cell progression; it sometimes results in a large stimulation of thymidine incorporation. When this concentration is applied to cells that have been irradiated with 1-krad doses of 220-kV x rays, there is a marked suppression of both the inhibition of DNA synthesis and G2 arrest induced by the radiation. Larger doses require higher concentrations of caffeine to suppress the inhibition of DNA synthesis. Delaying addition until the rate of synthesis is at its minimum (1.5 hours after irradiation with 1 krad) results in a slightly accelerated recovery of the rate. Treatment before or during irradiation is without effect on the inhibition. Removal of the caffeine as late as 6 hours after its addition at the time of irradiation results in a prompt inhibition in DNA synthesis that mimics that observed immediately after irradiation in the absence of caffeine. These findings raise the possibility that the depression in rate of DNA systhesis might not result from radiation damage introduced into the replicon initiation system, but rather may be an indirect consequence of damage residing elsewhere in the irradiated cell

333

Action of caffeine on x-irradiated HeLa cells. I. Delayed inhibition of DNA synthesis  

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Treatment of HeLa S3 cells with 1 mM caffeine delays progression through G1 by 1.5 hours but causes no other detectable inhibition of cell progression; it sometimes results in a large stimulation of thymidine incorporation. When this concentration is applied to cells that have been irradiated with 1-krad doses of 220-kV x rays, there is a marked suppression of both the inhibition of DNA synthesis and G2 arrest induced by the radiation. Larger doses require higher concentrations of caffeine to suppress the inhibition of DNA synthesis. Delaying addition until the rate of synthesis is at its minimum (1.5 hours after irradiation with 1 krad) results in a slightly accelerated recovery of the rate. Treatment before or during irradiation is without effect on the inhibition. Removal of the caffeine as late as 6 hours after its addition at the time of irradiation results in a prompt inhibition in DNA synthesis that mimics that observed immediately after irradiation in the absence of caffeine. These findings raise the possibility that the depression in rate of DNA systhesis might not result from radiation damage introduced into the replicon initiation system, but rather may be an indirect consequence of damage residing elsewhere in the irradiated cell.

Tolmach, L.J.; Jones, R.W.; Busse, P.M.

1977-09-01

334

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

Science.gov (United States)

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

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

2011-09-01

335

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

OpenAIRE

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

Barh, D.; Viswanathan, G.

2008-01-01

336

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

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

de Mello Samanta M

2008-12-01

337

Quantitative gene expression assessment identifies appropriate cell line models for individual cervical cancer pathways  

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Full Text Available Abstract Background Cell lines have been used to study cancer for decades, but truly quantitative assessment of their performance as models is often lacking. We used gene expression profiling to quantitatively assess the gene expression of nine cell line models of cervical cancer. Results We find a wide variation in the extent to which different cell culture models mimic late-stage invasive cervical cancer biopsies. The lowest agreement was from monolayer HeLa cells, a common cervical cancer model; the highest agreement was from primary epithelial cells, C4-I, and C4-II cell lines. In addition, HeLa and SiHa cell lines cultured in an organotypic environment increased their correlation to cervical cancer significantly. We also find wide variation in agreement when we considered how well individual biological pathways model cervical cancer. Cell lines with an anti-correlation to cervical cancer were also identified and should be avoided. Conclusion Using gene expression profiling and quantitative analysis, we have characterized nine cell lines with respect to how well they serve as models of cervical cancer. Applying this method to individual pathways, we identified the appropriateness of particular cell lines for studying specific pathways in cervical cancer. This study will allow researchers to choose a cell line with the highest correlation to cervical cancer at a pathway level. This method is applicable to other cancers and could be used to identify the appropriate cell line and growth condition to employ when studying other cancers.

Iyer Vishwanath R

2007-05-01

338

Cell phones and cancer  

Science.gov (United States)

Cancer and cell phones; Do cell phones cause cancer? ... Several major studies show no link between cell phones and cancer at this time. However, since the information available is based on short-term studies, the impact of many years of ...

339

Arsenic trioxide inhibits cell proliferation and human papillomavirus oncogene expression in cervical cancer cells.  

Science.gov (United States)

Arsenic trioxide (As2O3) has shown therapeutic effects in some leukemias and solid cancers. However, the molecular mechanisms of its anticancer efficacy have not been clearly elucidated, particularly in solid cancers. Our previous data showed that As2O3 induced apoptosis of human papillomavirus (HPV) 16 DNA-immortalized human cervical epithelial cells and cervical cancer cells and inhibited the expression of HPV oncogenes in these cells. In the present study, we systemically examined the effects of As2O3 on five human cervical cancer cell lines and explored the possible molecular mechanisms. MTT assay showed that HPV-negative C33A cells were more sensitive to growth inhibition induced by As2O3 than HPV-positive cervical cancer cells, and HPV 18-positive HeLa and C4-I cells were more sensitive to As2O3 than HPV 16-positive CaSki and SiHa cells. After As2O3 treatment, both mRNA and protein levels of HPV E6 and E7 obviously decreased in all HPV positive cell lines. In contrast, p53 and Rb protein levels increased in all tested cell lines. Transcription factor AP-1 protein expression decreased significantly in HeLa, CaSki and C33A cells with ELISA method. These results suggest that As2O3 is a potential anticancer drug for cervical cancer. PMID:25117446

Wang, Hongtao; Gao, Peng; Zheng, Jie

2014-09-01

340

Entry of human rhinovirus 89 via ICAM-1 into HeLa epithelial cells is inhibited by actin skeleton disruption and by bafilomycin.  

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HRV89, a major-group rhinovirus, uses intercellular adhesion molecule 1 (ICAM-1) for cell entry, while minor-group HRV2 uses the LDL receptor for clathrin-mediated endocytosis. Entry of HRV89 into HeLa epithelial cells was found to be inefficient, and infectious virus was still detected on the plasma membrane after 3 h of incubation with the cells. Endocytosis, and consequently infection, of HRV89 but not of HRV2, was almost completely blocked by the actin-polymerization inhibitor cytochalasin D, while the phosphatidylinositol 3-kinase inhibitor LY294002 had no effect on infection with either virus. Cytochalasin D also inhibited major-group HRV infection of rhabdomyosarcoma cells expressing ICAM-1 when the time available for uncoating was limited to 30 min. Although cholesterol depletion strongly inhibited HRV89 infection of HeLa cells, it only slightly affected HRV89 endocytosis, indicating that a lipid raft environment was not essential for virus uptake. The sodium-proton exchange inhibitor 5-(N-ethyl-N-isopropyl) amiloride (EIPA) significantly reduced cell entry and infection by HRV89 only at a concentration that also inhibited HRV2 infection and Alexa 488-transferrin entry. These data rule out classical macropinocytosis as an infectious entry pathway of HRV89 in HeLa cells. Notably, the proton ATPase inhibitor bafilomycin strongly affected cell entry of both viruses, suggesting a role for submembraneous pH in rhinovirus endocytosis. PMID:23913188

Pfanzagl, Beatrix; Andergassen, Daniel; Edlmayr, Johanna; Niespodziana, Katarzyna; Valenta, Rudolf; Blaas, Dieter

2014-01-01

341

Ginsenoside?Rg5 induces apoptosis and DNA damage in human cervical cancer cells.  

Science.gov (United States)

Panax ginseng is traditionally used as a remedy for cancer, inflammation, stress and aging, and ginsenoside?Rg5 is a major bioactive constituent of steamed ginseng. The present study aimed to evaluate whether ginsenoside?Rg5 had any marked cytotoxic, apoptotic or DNA?damaging effects in human cervical cancer cells. Five human cervical cancer cell lines (HeLa, MS751, C33A, Me180 and HT?3) were used to investigate the cytotoxicity of ginsenoside?Rg5 using a 3?(4,5?dimethylthiazol?2?yl)?2,5?diphenyltetrazolium bromide assay. Additionally, the effects of ginsenoside?Rg5 on the apoptosis of HeLa and MS751 cells were detected using DNA ladder assays and flow cytometry. DNA damage was assessed in the HeLa and MS751 cells using alkaline comet assays and by detection of ?H2AX focus formation. The HeLa and MS751 cells were significantly more sensitive to ginsenoside?Rg5 treatment compared with the C?33A, HT?3 and Me180 cells. As expected, ginsenoside?Rg5 induced significant concentration? and time?dependent increases in apoptosis. In addition, ginsenoside?Rg5 induced significant concentration?dependent increases in the level of DNA damage compared with the negative control. Consistent with the comet assay data, the percentage of ?H2AX?positive HeLa and MS751 cells also revealed that ginsenoside?Rg5 caused DNA double?strands to break in a concentration?dependent manner. In conclusion, ginsenoside?Rg5 had marked genotoxic effects in the HeLa and MS751 cells and, thus, demonstrates potential as a genotoxic or cytotoxic drug for the treatment of cervical cancer. PMID:25355274

Liang, Li-Dan; He, Tao; Du, Ting-Wei; Fan, Yong-Gang; Chen, Dian-Sen; Wang, Yan

2015-02-01

342

Ginsenoside-Rg5 induces apoptosis and DNA damage in human cervical cancer cells  

Science.gov (United States)

Panax ginseng is traditionally used as a remedy for cancer, inflammation, stress and aging, and ginsenoside-Rg5 is a major bioactive constituent of steamed ginseng. The present study aimed to evaluate whether ginsenoside-Rg5 had any marked cytotoxic, apoptotic or DNA-damaging effects in human cervical cancer cells. Five human cervical cancer cell lines (HeLa, MS751, C33A, Me180 and HT-3) were used to investigate the cytotoxicity of ginsenoside-Rg5 using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Additionally, the effects of ginsenoside-Rg5 on the apoptosis of HeLa and MS751 cells were detected using DNA ladder assays and flow cytometry. DNA damage was assessed in the HeLa and MS751 cells using alkaline comet assays and by detection of ?H2AX focus formation. The HeLa and MS751 cells were significantly more sensitive to ginsenoside-Rg5 treatment compared with the C-33A, HT-3 and Me180 cells. As expected, ginsenoside-Rg5 induced significant concentration- and time-dependent increases in apoptosis. In addition, ginsenoside-Rg5 induced significant concentration-dependent increases in the level of DNA damage compared with the negative control. Consistent with the comet assay data, the percentage of ?H2AX-positive HeLa and MS751 cells also revealed that ginsenoside-Rg5 caused DNA double-strands to break in a concentration-dependent manner. In conclusion, ginsenoside-Rg5 had marked genotoxic effects in the HeLa and MS751 cells and, thus, demonstrates potential as a genotoxic or cytotoxic drug for the treatment of cervical cancer. PMID:25355274

LIANG, LI-DAN; HE, TAO; DU, TING-WEI; FAN, YONG-GANG; CHEN, DIAN-SEN; WANG, YAN

2015-01-01

343

Celecoxib's radiosensitizing effects on three different human cancer cell lines  

International Nuclear Information System (INIS)

Objective: To investigate the radiosensitivity enhancement of celecoxib, a selective cyclooxygenase (COX-2) inhibitor, on three human cancer cell lines in vitro (the human lung adenocarcinoma cell lines A549, the human cervical carcinoma cell lines HeLa and the human nasopharyngeal carcinoma CNE). Methods: The subtoxic doses of celecoxib were chosen to do the radiosensitive experiment to A549 cells, HeLa cells and CNE cells. The three cells were divided into four groups: (1) the control (C); (2) the drug only (D); (3) the radiation only(R); (4) the radiation and drug (R + D). Celecoxib's radiosensitization on the three kinds of cells was measured by clongenic assay. Results: Celecoxib showed radiosensitizing effect on the three kinds of cells. The values of D0, Dq, SF2 and D0.01 of group R + D were significantly decreased in the group R. After A549 cell lines, HeLa cell lines, CNE cell lines were exposed to 30 ?mol/L celecoxib, SERD0 and SERDq were 1.26 and 1.34, 1.25 and 1.33, 1.24 and 1.32; respectively, in the group in which 50 ?mol/L celecoxib was used combined with radiation were 1.74 and 1.84, 1.36 and 1.47, 1.33 and 1.61. Conclusion: The subtoxic doses of celecoxib can enhance a concentration-dependent radiosensitivity of A549 cells, HeLa cells and CNE cells in vitro. Celecoxib may be widespread clinical used as a radiosensitizer. (authors)

344

Binding of host extracellular matrix proteins to Mycoplasma fermentans and its effect on adherence to, and invasion of HeLa cells.  

Science.gov (United States)

In the present study, we show that intact Mycoplasma fermentans cells have a wealth of adhesive interactions with components of the extracellular matrix. Mycoplasma fermentans intensively bind plasminogen, and to a lesser extent, fibronectin, heparin, and laminin. The binding of collagen type III, IV, or V was low. The binding of plasminogen, collagen type III, or collagen type V markedly enhanced the adherence of M. fermentans to HeLa cells, whereas the binding of fibronectin, heparin, laminin, or collagen IV induced only a small effect on mycoplasma adherence. Utilizing plasminogen-treated M. fermentans preparations, we detected microorganisms within host HeLa cells by the gentamicin protection assay or by confocal laser scanning microscopy of immunofluorescent preparations. However, no intracellular M. fermentans was detected when M. fermentans preparations treated with fibronectin, heparin, laminin, or collagen type III, IV, or V were utilized. PMID:17233726

Yavlovich, Amichai; Rottem, Shlomo

2007-01-01

345

Possible attenuation of the G2 DNA damage cell cycle checkpoint in HeLa cells by extremely low frequency (ELF electromagnetic fields  

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Full Text Available Abstract Background The issue remains unresolved as to whether low frequency magnetic fields can affect cell behaviour, with the possibility that they may be in part responsible for the increased incidence of leukaemia in parts of the population exposed to them. Methods Combined treatment of HeLa cells with gamma-irradiation (1, 3 and 5 Grays and extra low frequency magnetic fields of ~50 Hz was carried out under rigorously controlled conditions. Results Synchronised cells progressing from S-phase arrived at mitosis on average marginally ahead of irradiation controls not exposed to ELF. In no instance out of a total of twenty separate experiments did this "double-insult" further delay entry of cells into mitosis, as had been anticipated. Conclusion This apparently "non-genotoxic" agent (ELF appears to be capable of affecting cells that would normally arrest for longer in G2, suggesting a weakening of the stringency of the late cycle (G2 checkpoint.

Heaton Brian

2002-05-01

346

Association to HeLa cells and surface behavior of exogenous gangliosides studied with a fluorescent derivative of GM1  

International Nuclear Information System (INIS)

Cultured HeLa cells were incubated with pyrene-GM1/3H-radiolabeled GM1 ganglioside (1:4 M/M) mixtures for various times. The process of association of pyrene-GM1 with cells was qualitatively and quantitatively the same as that of 3H-GM1. The pyrene-GM1 and 3H-GM1 proportions in the various forms of association with cells were similar to that of the starting ganglioside mixture. After 2-h incubation, the association of ganglioside with cells was well established whereas almost no metabolic processing had occurred. During a 24-h incubation, pyrene- and 3H-GM1 underwent similar metabolic processing and gave rise to catabolic (GM2 and GM3) and anabolic (GDla) derivatives. Fluorescence spectroscopy experiments carried out with the excimer formation technique on subcellular fractions containing plasma membranes showed that exogenous ganglioside was, in part, associated with the cells in a micellar form removable by trypsin treatment, and in part inserted in a seemingly molecular dispersion. Addition of Ca2+ salts caused aggregation of the ganglioside, as indicated by the increase of the excimer:monomer fluorescence ratio. The phenomenon was Ca2+ concentration dependent (maximum at 10 mM), and subsequent addition of EDTA has no effect. The saccharide portion of exogenously incorporated pyrene-GM1 was available to interact with external ligands, as shown by its ability to bind cholera toxin whose addition reducto bind cholera toxin whose addition reduced the collision rate among the ganglioside lipid moieties

347

Lung Cancer Stem Cells  

OpenAIRE

Lung cancer remains a major cause of cancer-related lethality because of high incidence and recurrence in spite of significant advances in staging and therapies. Recent data indicates that stem cells situated throughout the airways may initiate cancer formation. These putative stem cells maintain protumorigenic characteristics including high proliferative capacity, multipotent differentiation, drug resistance and long lifespan relative to other cells. Stem cell signaling and differentiation p...

Pine, Sharon R.; Blair Marshall; Lyuba Varticovski

2008-01-01

348

A nucleic-acid hydrolyzing single chain antibody confers resistance to DNA virus infection in hela cells and C57BL/6 mice.  

Science.gov (United States)

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

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

2014-06-01

349

Comparison of the killing effects between nitrogen-doped and pure TiO2 on HeLa cells with visible light irradiation.  

Science.gov (United States)

The killing effect of nitrogen-doped titanium dioxide (N-TiO2) nanoparticles on human cervical carcinoma (HeLa) cells by visible light photodynamic therapy (PDT) was higher than that of TiO2 nanoparticles. To study the mechanism of the killing effect, the reactive oxygen species produced by the visible-light-activated N-TiO2 and pure-TiO2 were evaluated and compared. The changes of the cellular parameters, such as the mitochondrial membrane potential (MMP), intracellular Ca2+, and nitrogen monoxide (NO) concentrations after PDT were measured and compared for N-TiO2- and TiO2-treated HeLa cells. The N-TiO2 resulted in more loss of MMP and higher increase of Ca2+ and NO in HeLa cells than pure TiO2. The cell morphology changes with time were also examined by a confocal microscope. The cells incubated with N-TiO2 exhibited serious distortion and membrane breakage at 60 min after the PDT. PMID:23433090

Li, Zheng; Pan, Xiaobo; Wang, Tianlong; Wang, Pei-Nan; Chen, Ji-Yao; Mi, Lan

2013-01-01

350

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

International Nuclear Information System (INIS)

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

351

ANTICANCER ACTIVITY OF PONGAMIA GLABRA V. SEED OIL EXTRACT AGAINST SELECTED HUMAN CANCER CELL LINES  

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Full Text Available Screening of the seed oil extract from Pongamia glabra V. (Fabaceae has been carried out for antiproliferative activity of cancer cells. The seed oil was extracted with methanol and then persuasive activity was tested on human cancer cell lines MCF-7 and HeLa. The cell growth inhibitory effects of seed oil extract was observed. The cell viability was assessed using trypan blue dye exclusion method and 3-(4, 5- Dimethyl thiazol-2yl-2, 5-dimethyltetrazolium bromide (MTT assay. The IC50 value of the methanolic seed oil extract against MCF-7 and HeLa was found to be 6 mg/ml and 6 mg/ml respectively after 48 hours of incubation. The P.glabra seed oil extract increased the proportion of DNA fragmentation in MCF-7 and HeLa cancer cell lines. Moreover, the inhibitory effect is correlated with DNA fragmentation. These results suggest that the P.glabra seed oil extract has an inhibitory effect on human cancer cell lines MCF-7 and HeLa.

Chinnasamy Arulvasu

2012-08-01

352

Compatibility of cancer cells with nanostructured oxidized porous silicon substrates  

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The attachment and long-term viability of three types of human cancer cell lines (glioma U87, breast cancer MDA-MB-231, and cervical cancer HeLa) onto nanostructured oxidized porous Si substrates is investigated. The porous layers are fabricated to give cylindrically-shaped structures with pore diameters in the tunable range of 10 to 150 nm by anodizing a heavily-doped p-type Si. The Alamar Blue viability assay and optical microscopy are employed to assess the attachment, viability and the morphology of the cells. The results show that cells remain viable and proliferate on all surfaces. The nano-architecture of the studied scaffolds does not exert a deleterious effect on cancer cells. Cell coverage levels comparable to standard culture preparations on tissue culture polystyrene are observed (copyright 2011 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

Zeidman, Tal; Parush, Ran; Massad, Na' ama [Department of Biotechnology and Food Engineering, Technion - Israel Institute of Technology, Haifa 32000 (Israel); Segal, Ester [Department of Biotechnology and Food Engineering, Technion - Israel Institute of Technology, Haifa 32000 (Israel); Russell Berrie Nanotechnology Institute, Technion - Israel Institute of Technology, Haifa 32000 (Israel)

2011-06-15

353

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

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

James Ayorinde Babatunde

2013-01-01

354

Loss of Protein Kinase PKR Expression in Human HeLa Cells Complements the Vaccinia Virus E3L Deletion Mutant Phenotype by Restoration of Viral Protein Synthesis?  

Science.gov (United States)

The E3L proteins encoded by vaccinia virus bind double-stranded RNA and mediate interferon resistance, promote virus growth, and impair virus-mediated apoptosis. Among the cellular proteins implicated as targets of E3L is the protein kinase regulated by RNA (PKR). To test in human cells the role of PKR in conferring the E3L mutant phenotype, HeLa cells stably deficient in PKR generated by an RNA interference-silencing strategy were compared to parental and control knockdown cells following infection with either an E3L deletion mutant (?E3L) or wild-type (WT) virus. The growth yields of WT virus were comparable in PKR-sufficient and -deficient cells. By contrast, the single-cycle yield of ?E3L virus was increased by nearly 2 log10 in PKR-deficient cells over the impaired growth in PKR-sufficient cells. Furthermore, virus-induced apoptosis characteristic of the ?E3L mutant in PKR-sufficient cells was effectively abolished in PKR-deficient HeLa cells. The viral protein synthesis pattern was altered in ?E3L-infected PKR-sufficient cells, characterized by an inhibition of late viral protein expression, whereas in PKR-deficient cells, late protein accumulation was restored. Phosphorylation of both PKR and the ? subunit of protein synthesis initiation factor 2 (eIF-2?) was elevated severalfold in ?E3L-infected PKR-sufficient, but not PKR-deficient, cells. WT virus did not significantly increase PKR or eIF-2? phosphorylation in either PKR-sufficient or -deficient cells, both of which supported efficient WT viral protein production. Finally, apoptosis induced by infection of PKR-sufficient HeLa cells with ?E3L virus was blocked by a caspase antagonist, but mutant virus growth was not rescued, suggesting that translation inhibition rather than apoptosis activation is a principal factor limiting virus growth. PMID:17959656

Zhang, Ping; Jacobs, Bertram L.; Samuel, Charles E.

2008-01-01

355

Visualizing the effect of tumor microenvironments on radiation-induced cell kinetics in multicellular spheroids consisting of HeLa cells  

Energy Technology Data Exchange (ETDEWEB)

Highlights: •We visualized radiation-induced cell kinetics in spheroids. •HeLa-Fucci cells were used for detection of cell-cycle changes. •Radiation-induced G2 arrest was prolonged in the spheroid. •The inner and outer cell fractions behaved differently. -- Abstract: In this study, we visualized the effect of tumor microenvironments on radiation-induced tumor cell kinetics. For this purpose, we utilized a multicellular spheroid model, with a diameter of ?500 ?m, consisting of HeLa cells expressing the fluorescent ubiquitination-based cell-cycle indicator (Fucci). In live spheroids, a confocal laser scanning microscope allowed us to clearly monitor cell kinetics at depths of up to 60 ?m. Surprisingly, a remarkable prolongation of G2 arrest was observed in the outer region of the spheroid relative to monolayer-cultured cells. Scale, an aqueous reagent that renders tissues optically transparent, allowed visualization deeper inside spheroids. About 16 h after irradiation, a red fluorescent cell fraction, presumably a quiescent G0 cell fraction, became distinct from the outer fraction consisting of proliferating cells, most of which exhibited green fluorescence indicative of G2 arrest. Thereafter, the red cell fraction began to emit green fluorescence and remained in prolonged G2 arrest. Thus, for the first time, we visualized the prolongation of radiation-induced G2 arrest in spheroids and the differences in cell kinetics between the outer and inner fractions.

Kaida, Atsushi; Miura, Masahiko, E-mail: masa.mdth@tmd.ac.jp

2013-10-04

356

Ubiquitin B in Cervical Cancer: Critical for the Maintenance of Cancer Stem-Like Cell Characters  

Science.gov (United States)

Cervical cancer cells exhibit an increased requirement for ubiquitin-dependent protein degradation associated with an elevated metabolic turnover rate. Ubiquitin, which is a small, highly conserved protein expressed in all eukaryotic cells, can be covalently linked to certain target proteins to mark them for degradation by the ubiquitin-proteasome system. Previous studies highlight the essential role of Ubiquitin B (UbB) and UbB-dependent proteasomal protein degradation in histone deacetylase inhibitor (HDACi) -induced tumor selectivity. We hypothesized that UbB plays a critical role in the function of cervical cancer stem cells. We measured endogenous UbB levels in mammospheres in vitro by real-time PCR and Western blotting. The function of UbB in cancer stem-like cells was assessed after knockdown of UbB expression in prolonged Trichostatin A-selected HeLa cells (HeLa/TSA) by measuring in vitro cell proliferation, cell apoptosis, invasion, and chemotherapy resistance as well as by measuring in vivo growth in an orthotopic model of cervical cancer. We also assessed the cancer stem cell frequency, tumorsphere formation, and in vivo growth of human cervical cancer xenografts after UbB silencing. We found that HeLa/TSA were resistant to chemotherapy, highly expressed the UbB gene and the stem cell markers Sox2, Oct4 and Nanog. These cells also displayed induced differentiation abilities, including enhanced migration/invasion/malignancy capabilities in vitro and in vivo. Furthermore, an elevated expression of UbB was shown in the tumor samples of chemotherapy patients. Silencing of UbB inhibited tumorsphere formation, lowered the expression of stem cell markers and decreased cervical xenograft growth. Our results demonstrate that UbB was significantly increased in prolonged Trichostatin A-selected HeLa cells and it played a key role in the maintenance of cervical cancer stem-like cells. PMID:24367661

Wang, Yingying; Ji, Teng; Sun, Shujuan; Mo, Qingqing; Chen, Pingbo; Fang, Yong; Liu, Jia; Wang, Beibei; Zhou, Jianfeng; Ma, Ding; Wu, Peng

2013-01-01

357

Correlation between ?-ray-induced G2 arrest and radioresistance in two human cancer cells  

International Nuclear Information System (INIS)

Purpose: The correlation between radioresistance and ?-ray-induced G2 arrest was examined in two human cancer cell lines, HeLa (cervical carcinoma) and MeWo (melanoma). Methods and Materials: Cellular radioresistance was examined by a colony formation assay and Hoechst 33342 staining. G2 arrest induced by ?-rays was examined by flow cytometry, and the accumulation of cyclin B1 and cdc2 proteins was analyzed using Western blotting. Results: HeLa was more resistant (10% survival dose[D10] 10 Gy) than MeWo (D10 = 4 Gy) to ?-rays. In HeLa, cell cycle analysis showed that G2 arrest was induced 10 or 24 h after irradiation of 10 or 4 Gy, respectively. In contrast, no clear G2 arrest in MeWo was observed after irradiation. Western blot analysis showed that cell cycle regulators, cyclin B1 and cdc2, were accumulated in HeLa but not in MeWo. The accumulation of cyclin B1 and cdc2 reached peak levels 24-34 h after irradiation of 10 Gy, and 24 h after irradiation of 4 Gy. In addition, Hoechst staining revealed similar increase in apoptotic bodies with time after irradiation in HeLa and MeWo at isosurvival doses. Conclusion: Radioresistance of these human cancer cells is closely correlated with ?-ray-induced G2 arrest, and cyclin B1 and cdc2 are possible regulators of G2 arrest

358

Effects of activated aflatoxin B1 and caffeine on DNA replicon initiation in HeLa cells  

International Nuclear Information System (INIS)

Afatoxin B1 (AFB1) is activated by a rat microsomal extract (S-9) to form a product that inhibits DNA synthesis in HeLa cells. At 10-7 M, AFB1 inhibited initiation of replicons, as shown in alkaline sucrose gradient profiles 30 min after incubation with the drug. Ninety minutes later, the profile of treated cells was similar to that of control, but 4 h later there was another effect on replicon initiation. At 10-6 M, the inhibition of initiation was greater than at 10-7 M and increased progressively. Four hours after removal of the drug, the gradient profile showed low amounts of radioactivity in all size classes of DNA. When cells were incubated in medium containing caffeine (2 mM) even as late as 60 min after incubation with AFB1, the inhibition of replicon initiation was prevented. If caffeine was later removed from the medium, replicon initiation was then inhibited. At 10-7 M or 10-6 M, AFB1 had little immediate effect on chain elongation, but at 10-5 M, the gradient profiles showed an accumulation of low molecular weight DNA molecules, with no radioactivity in the region of high molecular weight DNA, owing to a block to chain elongation; this was not affected by caffeine. These results suggest that AFB1 induces damage that changes the fonformation of chromatin so that ges the fonformation of chromatin so that initiation of new replicons cannot occur; in the presence of caffeine this change does not occur and DNA replication is not inhibited

359

Extracellular gentamicin reduces the activity of connexin hemichannels and interferes with purinergic Ca2+ signaling in HeLa cells  

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Full Text Available Gap junction channels (GJCs and hemichannels (HCs are composed of protein subunits termed connexins (Cxs and are permeable to ions and small molecules. In most organs, GJCs communicate the cytoplasm of adjacent cells, while HCs communicate the intra and extracellular compartments. In this way, both channel types coordinate physiological responses of cell communities. Cx mutations explain several genetic diseases, including about 50% of autosomal recessive nonsyndromic hearing loss. However, the possible involvement of Cxs in the etiology of acquired hearing loss remains virtually unknown. Factors that induce post-lingual hearing loss are diverse, exposure to gentamicin an aminoglycoside antibiotic, being the most common. Gentamicin has been proposed to block GJCs, but its effect on HCs remains unknown. In this work, the effect of gentamicin on the functional state of HCs was studied and its effect on GJCs was reevaluated in HeLa cells stably transfected with Cxs. We focused on Cx26 because it is the main Cx expressed in the cochlea of mammals where it participates in purinergic signaling pathways. We found that gentamicin applied extracellularly reduces the activity of HCs, while dye transfer across GJCs was not affected. HCs were also blocked by streptomycin, another aminoglycoside antibiotic. Gentamicin also reduced the ATP release and the HC-dependent oscillations of cytosolic free-Ca2+ signal. Moreover, gentamicin drastically reduced the Cx26 HC-mediated membrane currents in Xenopus laevis oocytes. Therefore, the extracellular gentamicin-induced inhibition of Cx HCs may adversely affect autocrine and paracrine signaling, including the purinergic one, which might partially explain its ototoxic effects.

Vania A Figueroa

2014-09-01

360

Mistargeting of the Lectin ERGIC-53 to the Endoplasmic Reticulum of HeLa Cells Impairs the Secretion of a Lysosomal Enzyme  

OpenAIRE

ERGIC-53, a homo-oligomeric recycling protein associated with the ER–Golgi intermediate compartment (ERGIC), has properties of a mannose-selective lectin in vitro, suggesting that it may function as a transport receptor for glycoproteins in the early secretory pathway. To investigate if ERGIC-53 is involved in glycoprotein secretion, a mutant form of this protein was generated that is incapable of leaving the ER. If expressed in HeLa cells in a tetracycline-inducible manner, this muta...

Vollenweider, Florence; Kappeler, Felix; Itin, Christian; Hauri, Hans-peter

1998-01-01

361

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

OpenAIRE

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

Jagetia, Ganesh Chandra; Rao, Shaival Kamalaksha

2006-01-01

362

Increased protein secretion and adherence to HeLa cells by Shigella spp. following growth in the presence of bile salts.  

OpenAIRE

Growth of Shigella spp. in the presence of the bile salt deoxycholate or chenodeoxycholate enhanced the bacterial invasion of HeLa cells. Growth in the presence of other structurally similar bile salts or detergents had little or no effect. Deoxycholate-enhanced invasion was not observed when bacteria were exposed to deoxycholate at low temperatures or when chloramphenicol was added to the growth medium, indicating that bacterial growth and protein synthesis are required. Increased invasion i...

Pope, L. M.; Reed, K. E.; Payne, S. M.

1995-01-01

363

Effect of troglitazone on radiation sensitivity in cervix cancer cells  

International Nuclear Information System (INIS)

Troglitazone (TRO) is a peroxisome proliferator-activated receptor ? (PPAR? ) agonist. TRO has antiproliferative activity on many kinds of cancer cells via G1 arrest. TRO also increases Cu2+/Zn2+ -superoxide dismutase (CuZnSOD) and catalase. Cell cycle, and SOD and catalase may affect on radiation sensitivity. We investigated the effect of TRO on radiation sensitivity in cancer cells in vitro. Three human cervix cancer cell lines (HeLa, Me180, and SiHa) were used. The protein expressions of SOD and catalase, and catalase activities were measured at 2-10 ?M of TRO for 24 hours. Cell cycle was evaluated with flow cytometry. Reactive oxygen species (ROS) was measured using 2',7'-dichlorofluorescin diacetate. Cell survival by radiation was measured with clonogenic assay. By 5 ?M TRO for 24 hours, the mRNA, protein expression and activity of catalase were increased in all three cell lines. G0- G1 phase cells were increased in HeLa and Me180 by 5 ?M TRO for 24 hours, but those were not increased in SiHa. By pretreatment with 5 ?M TRO radiation sensitivity was increased in HeLa and Me180, but it was decreased in SiHa. In Me180, with 2 ?M TRO which increased catalase but not increased G0-G1 cells, radiosensitization was not observed. ROS produced by radiation was decreased with TRO. TRO increases radiation sensitivity through G0-G1 arrest or decreases radiation sensitivity through catalasemediated ROS scavenging according to TRO dose or cell typesaccording to TRO dose or cell types. The change of radiation sensitivity by combined with TRO is not dependent on the PPAR ? expression level.

364

Carbon nanowall scaffold to control culturing of cervical cancer cells  

Science.gov (United States)

The effect of carbon nanowalls (CNWs) on the culturing rate and morphological control of cervical cancer cells (HeLa cells) was investigated. CNWs with different densities were grown using plasma-enhanced chemical vapor deposition and subjected to post-growth plasma treatment for modification of the surface terminations. Although the surface wettability of the CNWs was not significantly dependent on the CNW densities, the cell culturing rates were significantly dependent. Morphological changes of the cells were not significantly dependent on the density of CNWs. These results indicate that plasma-induced surface morphology and chemical terminations enable nanobio applications using carbon nanomaterials.

Watanabe, Hitoshi; Kondo, Hiroki; Okamoto, Yukihiro; Hiramatsu, Mineo; Sekine, Makoto; Baba, Yoshinobu; Hori, Masaru

2014-12-01

365

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

Scientific Electronic Library Online (English)

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

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

2007-03-01

366

A Unique Role for Heat Shock Protein 70 and Its Binding Partner Tissue Transglutaminase in Cancer Cell Migration*  

OpenAIRE

Cell migration is essential for several important biological outcomes and is involved in various developmental disorders and disease states including cancer cell invasiveness and metastasis. A fundamental step in cell migration is the development of a leading edge. By using HeLa carcinoma cells as an initial model system, we uncovered a surprising role for the heat shock protein 70 (Hsp70) and its ability to bind the protein cross-linking enzyme, tissue transglutaminase (tTG), in cancer cell ...

Boroughs, Lindsey K.; Antonyak, Marc A.; Johnson, Jared L.; Cerione, Richard A.

2011-01-01

367

Breast cancer stem cells  

OpenAIRE

Breast cancer stem cells (BCSCs) constitute a subpopulation of tumor cells that express stem cell-associated markers and have a high capacity for tumor generation in vivo. Identification of BCSCs from tumor samples or breast cancer cell lines has been based mainly on CD44+/CD24?/low or ALDH+ phenotypes. BCSCs isolation has allowed the analysis of the molecular mechanisms involved in their origin, self-renewal, differentiation into tumor cells, resistance to radiation therapy and chemotherap...

MatthewJNaylor

2013-01-01

368

Effects of combined X-radiation and UV-radiation on HeLa cells  

International Nuclear Information System (INIS)

A combined X-ray-UV irradiation was performed in nonsynchronized HeLa-cells. A pre-irradiation with UV-light, that reduced the survival rate to 42% and the following X-ray radiation yielded a similar dose-effect characteristic as with ordinary X-ray irradiation, only its shoulder was smaller. An additive radiation interaction with the cellular molecular structure was observed. A pre-irradiation with X-rays followed by step-wise UV-irradiation yielded a function similar to the UV-action curve but also with a narrower shoulder. A additive effect could be observed. One can conclude from this that in combined irradiation two interacting processes cause the death of the cells. The gene mutations caused by UV-light lead to cell death. X-rays however cause chromosome breaks, that in an unfavourable combination also lead to cell death. The DNA distorsion caused by the UV-light increases the possibility of misrepair. (orig.)

369

Assay of anti-cancer drug sensitivity of cells from human tumors  

International Nuclear Information System (INIS)

The ability of cells to incorporate labeled nucleic acid precursors into acid precipitable material was assessed. Hela-S3 cells were employed to avoid inherited variability and heterogenity of primary cultures of human tumors. Labeled nucleic acid precursors were used to detect the viability of cells. A logarithm of counts per minute of incorporated labeled precursors is in proportion to a logarithm of viable cell numbers. Multiplate was used to provide large numbers of replicate cultures. Monolayer Hela-S3 cells which were seeded 24 hours earlier were incubated for three hours with various concentrations of drugs. After removal of drugs, cells were cultured for one week. In this period, Hela-S3 cells with no drug treatment became almost confluent and mitoses occurred about four times. Labeled precursors were incubated for three hours, and incorporated 3H-thymidine was counted. A standard curve of incorporated labeled precursor counts and viable cell numbers was drawn for every assay. The density inhibition of labeled nucleic acid precursor incorporation can be checked and corrected with the standard curve, and viable cell numbers after drug exposure can be obtained from the standard curve. When percent survival is plotted against drug concentration, a sigmoid curve is obtained if the drug has dose dependent effect and then the 90% lethal dose (LD90) can be determined from the curve. LD90 was used as the index of anti-cancer LD90 was used as the index of anti-cancer drug sensitivity. If the drug has time dependent effect, percent survival of maximal inhibition was used as the index of drug sensitivity. (J.P.N.)

370

Effect of troglitazone on radiation sensitivity in cervix cancer cells  

Energy Technology Data Exchange (ETDEWEB)

Troglitazone (TRO) is a peroxisome proliferator-activated receptor {gamma} (PPAR{gamma} ) agonist. TRO has antiproliferative activity on many kinds of cancer cells via G1 arrest. TRO also increases Cu{sup 2+}/Zn{sup 2+} -superoxide dismutase (CuZnSOD) and catalase. Cell cycle, and SOD and catalase may affect on radiation sensitivity. We investigated the effect of TRO on radiation sensitivity in cancer cells in vitro. Three human cervix cancer cell lines (HeLa, Me180, and SiHa) were used. The protein expressions of SOD and catalase, and catalase activities were measured at 2-10 {mu}M of TRO for 24 hours. Cell cycle was evaluated with flow cytometry. Reactive oxygen species (ROS) was measured using 2',7'-dichlorofluorescin diacetate. Cell survival by radiation was measured with clonogenic assay. By 5 {mu}M TRO for 24 hours, the mRNA, protein expression and activity of catalase were increased in all three cell lines. G0- G1 phase cells were increased in HeLa and Me180 by 5 {mu}M TRO for 24 hours, but those were not increased in SiHa. By pretreatment with 5 {mu}M TRO radiation sensitivity was increased in HeLa and Me180, but it was decreased in SiHa. In Me180, with 2 {mu}M TRO which increased catalase but not increased G0-G1 cells, radiosensitization was not observed. ROS produced by radiation was decreased with TRO. TRO increases radiation sensitivity through G0-G1 arrest or decreases radiation sensitivity through catalasemediated ROS scavenging according to TRO dose or cell types. The change of radiation sensitivity by combined with TRO is not dependent on the PPAR {gamma} expression level.

An, Zheng Zhe; Liu, Xian Guang; Song, Hye Jin; Choi, Chi Hwan; Kim, Won Dong; Park, Woo Yoon [Chungbuk National University College of Medicine, Cheongju (Korea, Republic of); Yu, Jae Ran [Konkuk University College of Medicine, Chungju (Korea, Republic of)

2012-06-15

371

Time-resolved microfluorometric study of the binding sites of lipophilic cationic pyrene probes in mitochondria of living HeLa cells.  

Science.gov (United States)

Lipophilic dye cations specifically bind to the mitochondria of living cells. Using fluorescent dyes, the mitochondria can easily be observed with a fluorescence microscope. Electron microscopy has shown that the dyes are bound to the inner mitochondrial membranes and the cristae. Using time-resolved fluorescence microscopy we have investigated, whether the dye molecules are preferentially accumulated at the strongly hydrophobic protein complexes of energy metabolism or at the lipids of the inner membrane system. In order to use our nanosecond-pulsed laser fluorometer we synthesized specially designed lipophilic pyrene cations with S1 lifetimes in the nanosecond domain, which specifically stain mitochondria in living HeLa cells. Model experiments with artificial membranes such as liposomes, proteoliposomes and also protein complexes have shown that the fluorescence is strongly quenched by oxygen if the pyrene probes are bound to lipids. Binding to proteins causes a much smaller quenching effect. In artificial systems, all decays were single exponential. This is in contrast with incubated HeLa cells, which showed double-exponential fluorescence decays. Comparing these with the artificial systems we came to the conclusion that in HeLa cells the long-lived species 1 are pyrene probes preferentially bound to the proteins of the inner mitochondrial membranes. The short-lived species 2 is caused by fluorescence resonance energy transfer from the pyrene probes as donors to cytochromes of the inner membranes as acceptors. From our decay data we estimated a mean distance between donor and acceptor of about 40 A. This is the same order of magnitude as the mean diameters of several mitochondrial protein complexes. Therefore we assumed that species 2 are pyrene probes bound either to mitochondrial proteins with cytochromes as constituents or to the interface between these proteins and the phospholipids of the membranes. Thus both species 1 and species 2 are spatially related to mitochrondrial proteins. This agrees with the observation that respiration of HeLa cells as well as cytochrome c oxidase vesicles (COVs) are inhibited with increasing concentration of pyrene probes. Finally, we studied the photodynamic effect on irradiation of HeLa cells and of COVs after incubation with lipophilic pyrene and porphyrine cations. PMID:8583282

Hüglin, D; Seiffert, W; Zimmermann, H W

1995-12-01

372

Attempts to purify a second cellular receptor for a coxsackievirus B3 variant, CB3-RD from HeLa cells.  

Science.gov (United States)

A coxsackievirus B3 variant, CB3-RD, isolated on rhabdomyosarcoma (RD) cells is known to bind HeLa cells at two different receptor protein sites, HR1 and HR2. Since HR2 occurs in almost 50 fold excess of HR1 in HeLa cells, purification of HR2 was attempted, to obtain its partial N-terminal amino acid sequence and its further characterization. This study describes the purification of HR2 from octylthioglucoside solubilized HeLa cell membranes (HeLa-OTG) by preparative isoelectric focusing (IEF) followed by either preparative sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) or affinity chromatography on immobilized receptor monoclonal antibody, RmcA (RmcA-agarose). IEF of HeLA-OTG showed that both HR2 and HR1 could be well separated by this technique and focused with peak maxima around pH 3.7 and 6.7, respectively. Both RmcA and CB3-RD recognized HR2 as doublet bands (60 kD major polypeptide and a minor 55 kD polypeptide) on electroblots under non-reducing conditions. Preparative SDS-PAGE of the pool of IEF fractions containing HR2 (IEF pool) and simultaneous elution of polypeptides from the bottom of the gel during electrophoresis, is shown to be a useful technique in purifying HR2 with only one contaminating polypeptide (65 kD). However, affinity chromatography of the IEF pool on RmcA-agarose yielded HR2 without any detectable contaminating polypeptide. A quantitative chemiluminescence assay was developed to estimate the amount of HR2 on HeLa cells and in solution, when dot blotted on polyvinylidene difluoride (PVDF) membranes and probed with RmcA. Assays revealed that about 1.2% of the total HR2 present on HeLa cells could be obtained by IEF followed by affinity chromatography. Efforts are continuing to obtain sufficient quantities of purified HR2 for partial N-terminal amino acid sequencing. PMID:8237113

Mohanty, J G; Crowell, R L

1993-09-01

373

Enhanced specific absorption rate in silanol functionalized Fe3O4 core-shell nanoparticles: study of Fe leaching in Fe3O4 and hyperthermia in L929 and HeLa cells.  

Science.gov (United States)

Core-shell Fe3O4-SiO2 magnetic nanoparticles (MNPs) have been synthesized using a simple synthesis procedure at different temperatures. These MNPs are used to investigate the effect of surface coating on specific absorption rate (SAR) under alternating magnetic field. The temperature achieved by silica coated Fe3O4 is higher than that by uncoated MNPs (Fe3O4). This can be attributed to extent of increase in Brownian motion for silica coated MNPs. The sample prepared at optimized temperature of 80°C shows the highest SAR value of 111W/g. It is found that SAR value decreases with increase in shell thickness. The chemical stability of these samples is analyzed by leaching experiments at pH 2-7. The silica coated samples are stable up to 7 days even at pH 2. Biocompatibility of the MNPs is evaluated in vitro by assessing their cytotoxicity on L929 and human cervical cancer cells (HeLa cells) using sulforhodamine-B assay. Their hyperthermic killing ability is also evaluated in HeLa cells using the same method. Cells treated with MNPs along with induction heating show decrease in viability as compared to that without induction heating. Further, cell death is found to be ?55% more in cells treated with silica coated MNPs under induction heating as compared to untreated control. These results establish the efficacy of Fe3O4-SiO2 prepared at 80°C in killing of tumor cells by cellular hyperthermia. PMID:25089699

Majeed, Jerina; Pradhan, Lina; Ningthoujam, Raghumani Singh; Vatsa, Rajesh Kumar; Bahadur, Dhirendra; Tyagi, Avesh Kumar

2014-10-01

374

Azithromycin Synergistically Enhances Anti-Proliferative Activity of Vincristine in Cervical and Gastric Cancer Cells  

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Full Text Available In this study, the anti-proliferative and anticancer activity of azithromycin (AZM was examined. In the presence of AZM, cell growth was inhibited more effectively in Hela and SGC-7901 cancer cells, relative to transformed BHK-21 cells. The respective 50% inhibition of cell growth (IC50 values for Hela, SGC-7901 and BHK-21 were 15.66, 26.05 and 91.00 µg/mL at 72 h post incubation, indicative of a selective cytotoxicity against cancer cells. Cell apoptosis analysis using Hoechst nuclear staining and annexin V-FITC binding assay further demonstrated that AZM was capable of inducing apoptosis in both cancer cells and transformed cells. The apoptosis induced by AZM was partly through a caspase-dependent mechanism with an up-regulation of apoptotic protein cleavage PARP and caspase-3 products, as well as a down-regulation of anti-apoptotic proteins, Mcl-1, bcl-2 and bcl-X1. More importantly, a combination of AZM and a low dose of the common anti-cancer chemotherapeutic agent vincristine (VCR, produced a selectively synergistic effect on apoptosis of Hela and SGC-7901 cells, but not BHK-21 cells. In the presence of 12.50 ?g/mL of VCR, the respective IC50 values of Hela, SGC-7901 and BHK-21 cells to AZM were reduced to 9.47 µg/mL, 8.43 µg/mL and 40.15 µg/mL at 72 h after the incubation, suggesting that the cytotoxicity of AZM had a selective anti-