WorldWideScience
1

Targeting Hela cancer cells with avidin-dendrimer  

International Nuclear Information System (INIS)

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

2

Radiation sensitization by dihydroartemisinin on human HeLa cells of cervical cancer  

International Nuclear Information System (INIS)

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)

3

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

International Nuclear Information System (INIS)

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

4

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

International Nuclear Information System (INIS)

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)

5

Baicalein induces apoptosis of human cervical cancer HeLa cells in vitro.  

Science.gov (United States)

A number of studies have shown that baicalein shows high antitumor activity in vitro and in vivo. In this study, the inhibitory effect of baicalein on human cervical cancer HeLa cells was studied in vitro. HeLa cells were treated with high (100 µg/ml) and low (50 µg/ml) doses of baicalein, and cell growth inhibition rates were examined by the MTT assay. The morphological changes of apoptotic cells were observed under the light and electron microscope, while the rate of cell apoptosis was examined by flow cytometry. The expression of apoptosis-related proteins was analyzed by western blot, and caspase-3 activation was examined by a caspase-3 activity assay and spectrophotometry. The results demonstrated that baicalein inhibits the proliferation of HeLa cells and induces apoptosis in a caspase-3-dependent pathway, through downregulation of the B-cell lymphoma 2 (Bcl-2) protein and upregulation of the Bcl-2-associated X protein (Bax), Fas, Fas ligand (FasL) and caspase-8. Thus, we conclude that baicalein induces apoptosis of HeLa cells via the mitochondrial and the death receptor pathways. Cell apoptosis in HeLa cells was most likely promoted by the activation of the proteolytic enzyme caspase-3 in both pathways. PMID:25373554

Peng, Yong; Guo, Congshan; Yang, Yanhong; Li, Fenglin; Zhang, Yanxia; Jiang, Bin; Li, Qingwang

2015-03-01

6

Proteomic Investigation into Betulinic Acid-Induced Apoptosis of Human Cervical Cancer HeLa Cells  

Science.gov (United States)

Betulinic acid is a pentacyclic triterpenoid that exhibits anticancer functions in human cancer cells. This study provides evidence that betulinic acid is highly effective against the human cervical cancer cell line HeLa by inducing dose- and time-dependent apoptosis. The apoptotic process was further investigated using a proteomics approach to reveal protein expression changes in HeLa cells following betulinic acid treatment. Proteomic analysis revealed that there were six up- and thirty down-regulated proteins in betulinic acid-induced HeLa cells, and these proteins were then subjected to functional pathway analysis using multiple analysis software. UDP-glucose 6-dehydrogenase, 6-phosphogluconate dehydrogenase decarboxylating, chain A Horf6-a novel human peroxidase enzyme that involved in redox process, was found to be down-regulated during the apoptosis process of the oxidative stress response pathway. Consistent with our results at the protein level, an increase in intracellular reactive oxygen species was observed in betulinic acid-treated cells. The proteins glucose-regulated protein and cargo-selection protein TIP47, which are involved in the endoplasmic reticulum pathway, were up-regulated by betulinic acid treatment. Meanwhile, 14-3-3 family proteins, including 14-3-3? and 14-3-3?, were down-regulated in response to betulinic acid treatment, which is consistent with the decrease in expression of the target genes 14-3-3? and 14-3-3?. Furthermore, it was found that the antiapoptotic bcl-2 gene was down-regulated while the proapoptotic bax gene was up-regulated after betulinic acid treatment in HeLa cells. These results suggest that betulinic acid induces apoptosis of HeLa cells by triggering both the endoplasmic reticulum pathway and the ROS-mediated mitochondrial pathway. PMID:25148076

Xu, Tao; Pang, Qiuying; Zhou, Dong; Zhang, Aiqin; Luo, Shaman; Wang, Yang; Yan, Xiufeng

2014-01-01

7

HeLa human cervical cancer cell migration is inhibited by treatment with dibutyryl-cAMP.  

Science.gov (United States)

Cyclic AMP (cAMP) activates both protein kinase A (PKA) and guanine-nucleotide exchange factor exchange protein directly activated by CAMP (EPAC)-mediated Ras-related Protein1 (RAP1) GTPase that regulates various cellular functions including cell migration. Herein, we investigated whether cAMP-mediated PKA and EPAC1/RAP1 pathways differentially control HeLa cervical cancer cell migration. Although HeLa cell migration was reduced by dibutyryl-cAMP, we observed an increase in cAMP/PKA, cAMP/EPAC1/RAP1-GTPase, and RAC1-GTPase. HeLa cell migration and RAC1-GTPase were increased by treatment with 8-(4-chloro-phenylthio)-2'-O-methyladenosine-3',5'-cAMP analogue to activate EPAC-specific signaling pathways. When HeLa cells were treated with H-89, a PKA inhibitor, cell migration was enhanced but RAC1-GTPase was inhibited. In addition, cell migration induced by dibutyryl-cAMP was reversed but the activity of Rac1-GTPase was inhibited by H-89 treatment. Taken together, these data demonstrate that cAMP/PKA and cAMP/EPAC1/RAP1-GTPase might inversely control cervical cancer cell migration, although both signaling pathways may up-regulate RAC1-GTPase. It also suggests that cAMP-mediated cancer cell migration was independent of RAC1-GTPase activation. PMID:24982353

Lee, Jae-Wook; Lee, Jiyoung; Moon, Eun-Yi

2014-07-01

8

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

International Nuclear Information System (INIS)

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)

9

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

Science.gov (United States)

The possible biochemical activities of the acetonic/ethanolic extract of the leaves of Maytenus procumbens (L.M.P), and its isolated compounds were investigated in the present study. In cytotoxicity assay, L.M.P showed IC(50) of 68.79, 51.22, 78.49, 76.59, and 76.64?g/ml on Caco-2, HeLa, HT29, NIH3T3, and T47D cells, respectively. Bioassay guided fractionation led to the isolation and identification of a new triterpene: '30-hydroxy-11?-methoxy-18?-olean-12-en-3-one' (HMO) in addition to a known terpenoid: 'asiatic acid' (AA). HMO exhibited the most cytotoxicity against HeLa cells and was further investigated for its ability to induce apoptosis in HeLa cells. HMO induced apoptosis up to 20.41% in HeLa cells versus control group (0.40%). Antioxidant/oxidative properties of L.M.P and HMO were investigated using extracellular (DPPH), and intracellular (ROS) assays. Experimental samples represented a time and concentration-dependent formation of ROS in Hela cells. Generation of ROS seems one of the mechanisms by which HMO induces apoptosis in Hela cells. Conclusion is that the active components in L.M.P might serve as a mediator of the ROS scavenging system and have the potential to act as prooxidant or antioxidant depending on the biological environment of the cells. PMID:22989702

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

2013-01-01

10

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

Science.gov (United States)

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

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

2014-03-01

11

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

International Nuclear Information System (INIS)

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)

12

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

International Nuclear Information System (INIS)

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)

13

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

International Nuclear Information System (INIS)

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

14

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

Science.gov (United States)

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

15

Fucoxanthin induces apoptosis in human cervical cancer cell line HeLa via PI3K/Akt pathway.  

Science.gov (United States)

Cervical cancer (CC) is a malignant neoplasm arising from cells originating in the cervix uteri, among the top causes of death from cancer in women. In a gene expression profiling study of metabolic response to treatment, PI3K/Akt signaling pathway are associated with the development of CC. A common mechanism of Akt activation seen in cancer types is alterations in the upstream regulators of Akt such as phosphatidylinositol 3-kinase (PI3K), which is overexpressed in cervical cancer tissues, and leads to phosphorylation of Akt. Both PI3K and Akt inhibitors exist and may be therapeutically valuable. In the present study, we use MTT assay and western blot for the high-throughput screening to select specific inhibitors of PI3K/Akt signaling pathway, and then obtain fucoxanthin. Fucoxanthin is a water-soluble dietary fiber, taken from the unique slimy component of alginic cells. Various studies have pointed out that fucoxanthin is very effective for the treatment of cancer. Our results have shown that fucoxanthin induced a significant apoptosis of HeLa cells, compared with other candidates. After treatment with fucoxanthin for 24 h, the level of phosphorylation was inhibited in a dose-dependent manner, and the proteins of apoptotic markers were changed in HeLa cells. And fucoxanthin could suppress tumor growth in vivo. In addition, the mitochondrial signal transduction pathway maybe was involved in its mechanism and NF-?B activation was decreased after treatment with fucoxanthin. Therefore, fucoxanthin may be used as anti-cervical cancer drugs in the future. PMID:25113250

Ye, Guoliu; Lu, Qin; Zhao, Weidong; Du, Danli; Jin, Lijie; Liu, Yusheng

2014-11-01

16

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

OpenAIRE

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

Brito, Daniela A.; Rieder, Conly L.

2009-01-01

17

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

International Nuclear Information System (INIS)

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)

18

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

Directory of Open Access Journals (Sweden)

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

Yuan Li

2014-07-01

19

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

Science.gov (United States)

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

20

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

Directory of Open Access Journals (Sweden)

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

Smith Charlotte M

2013-01-01

21

DYTOGENETIC ANALYSIS OF HELA AND CHANG CELLS  

OpenAIRE

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

22

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

International Nuclear Information System (INIS)

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

23

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

OpenAIRE

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 cumulat...

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

24

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

OpenAIRE

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...

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

2010-01-01

25

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.

26

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

27

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

Science.gov (United States)

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

Rupachandra, S; Sarada, D V L

2014-01-01

28

High Cytotoxic Activity of Phosphonium Salts and Their Complementary Selectivity towards HeLa and K562 Cancer Cells: Identification of Tri-n-butyl-n-hexadecylphosphonium bromide as a Highly Potent Anti-HeLa Phosphonium Salt  

OpenAIRE

Quaternary ammonium and phosphonium salts have been screened for their toxic effect on HeLa and K562 cancer cell lines, as well as on normal HUVEC cells. Tri-n-butyl-n-hexadecylphosphonium bromide, the first phosphonium salt with a halogen anion tested against HeLa cells, was 12 times more potent (IC50 500 ?m after 24 and 48 h) against both HeLa and K562 cancer cell lines as well as normal HUVEC cells, showing that the nontoxic N+CH2YMe (Y=S, O) structural motif in ammonium salts could be su...

Bachowska, Barbara; Kazmierczak-baranska, Julia; Cieslak, Marcin; Nawrot, Barbara; Szcze?sna, Dorota; Skalik, Joanna; Ba?czewski, Piotr

2012-01-01

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

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

OpenAIRE

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...

Ryu, Yun-kyoung; Lee, Jae-wook; Moon, Eun-yi

2015-01-01

31

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

32

Anti-proliferation and induced mitochondria-mediated apoptosis of ganoderic acid Mk from Ganoderma lucidum mycelia in cervical cancer HeLa cells  

OpenAIRE

Ganoderic acid Mk (GA-Mk), a triterpenoid acid, was isolated from the mycelia of Ganoderma lucidum, and no biological activity of GA-Mk has ever been reported. In this work, we investigated the effect of GA-Mk on the cell proliferation and apoptosis in HeLa cells. The MTT results demonstrated that GA-Mk displayed interesting cytotoxicity toward various human cancer cell lines. Bromodeoxyuridine (BrdU) incorporation assay showed that GA-Mk had a dose-dependent inhibitory effect on proliferatio...

Zhong, Jian-jiang; Li, Ying-bo; Liu, Ru-ming

2012-01-01

33

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

Science.gov (United States)

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

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

2010-06-01

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.  

Science.gov (United States)

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

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

2013-08-01

36

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

Science.gov (United States)

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

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

2013-01-01

37

Synchronization of HeLa cells.  

Science.gov (United States)

HeLa is one of the oldest and most commonly used cell lines in biomedical research. Owing to the ease of which they can be effectively synchronized by various methods, HeLa cells have been used extensively for studies of the cell cycle. Here we describe several protocols for synchronization of HeLa cells from different phases of the cell cycle. Synchronization in G(1) phase can be achieved with the HMG-CoA reductase inhibitor lovastatin, S phase with a double thymidine block procedure, and G(2) phase with the CDK inhibitor RO3306. Cells can also be enriched in mitosis by treating with nocodazole and mechanical shake-off. Release of the cells from these blocks enables researchers to follow gene expression and other events through the cell cycle. We also describe several protocols, including flow cytometry, BrdU labeling, immunoblotting, and time-lapse microscopy, for validating the synchrony of the cells and monitoring the progression of the cell cycle after release. PMID:21755447

Ma, Hoi Tang; Poon, Randy Y C

2011-01-01

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

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

40

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

OpenAIRE

Natural products are being extensively explored for their potential to prevent as well as treat cancer due to their ability to target multiple molecular pathways. Ficus religiosa has been shown to exert diverse biological activities including apoptosis in breast cancer cell lines. In the present study, we report the anti-neoplastic potential of aqueous extract of F. religiosa (FRaq) bark in human cervical cancer cell lines, SiHa and HeLa. FRaq altered the growth kinetics of SiHa (HPV-16 posit...

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

2013-01-01

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

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

OpenAIRE

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 ...

Abbas Zare Mirakabadi; Ali Sarzaeem

2012-01-01

43

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

International Nuclear Information System (INIS)

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

44

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 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

45

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

Science.gov (United States)

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

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

2006-12-01

46

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

Science.gov (United States)

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

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

2014-11-01

47

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)

48

Anticancer activity of Bombyx batryticatus ethanol extract against the human tumor cell line HeLa.  

Science.gov (United States)

Anticancer activity of Bombyx batryticatus ethanol extract (BBE) against HeLa cells was studied using cell viability, DNA fragmentation, real-time polymerase chain reaction, and Western blot analyses. The BBE inhibited the growth and induced apoptosis of HeLa cells. The MTT assay indicated that the BBE induced cytotoxicity in HeLa cells in a time- and concentration-dependent manner. When HeLa cells were treated for 48 h, the 50% inhibitory concentration (IC50) value for the BBE was 1.564 mg/mL. The microscopy results showed that HeLa cells were severely distorted and showed slow growth; some cells became round in shape when treated with 5 mg/mL BBE for 24 h. The DNA ladder results revealed excessive DNA fragmentation in HeLa cells treated with 7 mg/mL BBE for 36 h. The proapoptotic activity of the BBE was attributed to its ability to modulate the expression of Bcl-2 and Bax genes. The mRNA and protein expression levels of Bax were remarkably higher whereas those of Bcl-2 were lower than those in the control cells; this led to an increased Bax/Bcl-2 ratio in cells treated with the BBE for 36 h. The results suggest that the BBE might play an important role in tumor growth suppression by inducing apoptosis in human cervical cancer cells via the regulation of the Bcl-2- and Bax-mediated apoptotic pathways. PMID:25729938

Wu, W P; Cao, J; Wu, J Y; Chen, H; Wang, D

2015-01-01

49

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

Directory of Open Access Journals (Sweden)

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

Wei ZHANG

2013-02-01

50

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

OpenAIRE

Objective: Cervical cancer is the second most common cancer as well as one of leading cause of cancer-related death for women worldwide. In regards to that issue, focus of this paper will be on popularly used Piperaceae members including Piper betle L, Piper cf fragile Benth, Piper umbellatum L, Piper aduncum L, Piper pellucidum L. This research was conducted to elucidate the antioxidant, anticancer and apoptosis inducing activities of Piperaceae extracts on cervical cancer cells, namely HeLa...

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

2013-01-01

51

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

Directory of Open Access Journals (Sweden)

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

Wojciech Król

2008-01-01

52

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

Directory of Open Access Journals (Sweden)

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

Syahida Ahmad

2012-09-01

53

Antiproliferative effects of some medicinal plants on HeLa cells  

Directory of Open Access Journals (Sweden)

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

54

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)

55

Methanol extract from Vietnamese Caesalpinia sappan induces apoptosis in HeLa cells  

Scientific Electronic Library Online (English)

Full Text Available BACKGROUND: This study evaluated the cytotoxic activity of extracts from Caesalpinia sappan heartwood against multiple cancer cell lines using an MTT cell viability assay. The cell death though induction of apoptosis was as indicated by DNA fragmentation and caspase-3 enzyme activation. RESULTS: A m [...] ethanol extract from C. sappan (MECS) showed cytotoxic activity against several of the cancer cell lines. The most potent activity exhibited by the MECS was against HeLa cells with an IC50value of 26.5?±?3.2 µg/mL. Treatment of HeLa cells with various MECS concentrations resulted in growth inhibition and induction of apoptosis, as indicated by DNA fragmentation and caspase-3 enzyme activation. CONCLUSION: This study is the first report of the anticancer properties of the heartwood of C. sappan native to Vietnam. Our findings demonstrate that C. sappan heartwood may have beneficial applications in the field of anticancer drug discovery.

Tran Manh, Hung; Nguyen Hai, Dang; Nguyen Tien, Dat.

56

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

Directory of Open Access Journals (Sweden)

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

Alcindo A. Dos Santos

2013-01-01

57

The effect of abraxane on cell kinetic parameters of HeLa cells.  

Science.gov (United States)

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

Gurses, Nurcan; Topcul, Mehmet

2013-01-01

58

Tumoricidal effects of nanomaterials in HeLa cell line  

Science.gov (United States)

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

59

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

60

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

61

Genetic transformation of HeLa cells by Agrobacterium  

OpenAIRE

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

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

2001-01-01

62

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

63

Studies on radioresistance with HeLa cells  

International Nuclear Information System (INIS)

In our previous experiments (15, 16), HeLa cells were successively irradiated with 1 kR of X-rays. After receiving 3, 5, 8, 11, 14, and 17 kR the iradiated HeLa cells showed progressively increasing radioresistance. In the present paper, the results of studies on the variation of the extrapolation number (n), mean lethal dose (D sub(o)), and quasi-threshold dose (D sub(q)) of survival curves for these radioresistant HeLa strains were reported. The values of n of the original and radioresistant strains were all found between 2 and 3, and a change of the n values with an increase in the total dose received by the cell strains was not noticed. On the other hand, the values of D sub(o) of the radioresistant strains progressively increased from 105 R in the original line to 148 R in the 17 kR strain with increasing doses of pre-irradiation. Similarly, the values of D sub(q) had a tendency to increase in the radioresistant strains. Repair of sublethal damage with the original and radioresistant strains was studied and the properties of the radioresistant strains were discussed. (author)

64

Inhibition of protein synthesis in intact HeLa cells.  

Science.gov (United States)

Polysome analysis has proved to be a sensitive probe for the mode of action of inhibitors of protein synthesis in intact HeLa cells. To classify the active compounds as inhibitors of initiation, elongation, or termination, their effects on the cellular polyribosome pattern were compared under three conditions. These conditions tested (i) their direct effect on the polyribosome profile; (ii) their effect on ribosome run-off produced by hypertonicity; and (iii) their effects on recovery from hypertonicity. Using this technique, diacetoxyscirpenol, 2-(4-methyl-2,6-dinitroanilino)-N-methylpropionamide, and three alkaloids, harringtonine, isoharringtonine, and homoharringtonine, were found to be inhibitors of initiation. Polysome analysis indicated that in HeLa cells 7.8 x 10(-7) M pactamycin, which inhibited protein synthesis 94%, interfered with elongation as well as initiation under these conditions. Emetine, anisomycin, cycloheximide, and trichodermin each gave polysome patterns consistent with inhibition of elongation. Fusidic acid and aurintricarboxylic acid inhibited incorporation of [(14)C]leucine into intact HeLa cells, but polysome analysis did not localize any specific inhibitory effects to the initiation, elongation, or termination steps of protein synthesis. The use of specific inhibitors of initiation of protein synthesis has indicated that most, if not all, mammalian messenger ribonucleic acids contain a single initiation site. PMID:1190754

Tscherne, J S; Pestka, S

1975-10-01

65

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

Science.gov (United States)

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

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

2014-09-01

66

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

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

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

69

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)

70

Outcome of Treatment of Human HeLa Cervical Cancer Cells With Roscovitine Strongly Depends on the Dosage and Cell Cycle Status Prior to the Treatment.  

Czech Academy of Sciences Publication Activity Database

Ro?. 106, ?. 5 (2009), s. 937-955. ISSN 0730-2312 Institutional research plan: CEZ:AV0Z50380511 Keywords : APOPTOSIS * CELL CYCLE ARREST * CYCLIN-DEPENDENT KINASES Subject RIV: ED - Physiology Impact factor: 2.935, year: 2009

Wesierska-Gadek, J.; Borza, A.; Walzi, E.; Kryštof, Vladimír; Maurer, M.; Komina, O.; Wandl, S.

2009-01-01

71

Effect of estrogens on bacterial adherence to HeLa cells.  

OpenAIRE

Incubating confluent cell culture HeLa cells for 18 h with increasing concentrations of estrogens progressively enhanced the subsequent attachment of a variety of radiolabeled bacteria to the HeLa cells. This effect was not caused by other hormones and was not produced by 1-h incubations of HeLa cells or bacteria with hormones. Estrogens did not similarly affect two other receptor cell lines studied. The addition of metabolic inhibitors showed that this effect of estrogens on HeLa cells was e...

Sugarman, B.; Epps, L. R.

1982-01-01

72

Requirement of E7 oncoprotein for viability of HeLa cells.  

Science.gov (United States)

Most human papillomavirus (HPV)-positive cervical cancers contain integrated copies of the viral genome in their chromosomes and express the viral oncoproteins E6 and E7. A virus-encoded transcription factor, E2, is known to repress E6/E7 expression in HPV-positive cancer cells, leading to growth inhibition, which indicates that E6/E7 is required for the survival of the cells. We found that the E2-mediated growth inhibition of HeLa cells, an HPV18-positive cancer cell line, was coupled with a reduction in telomerase activity, an effect which was rescued by the complementation of E7 expression, but not E6 expression, indicating that the cell viability and the telomerase activity in HeLa cells are maintained by an E7-associated function. Analysis of E7 mutants suggested that the binding to the pRB family of pocket proteins was involved in the ability of E7 to rescue the growth potential and telomerase activity inhibited by E2 expression. We also showed that the telomerase activity upregulated by E7 expression was determined by the hTERT promoter activity, and that c-Myc upregulation caused by pRB inactivation could account for the promoter activity. The activation of p53 and consequent accumulation of p21Cip1, which were triggered by the downregulation of E6, appeared not to be essential for the E2-mediated growth arrest. PMID:16500131

Nishimura, Akiko; Nakahara, Tomomi; Ueno, Takaharu; Sasaki, Kenta; Yoshida, Satoshi; Kyo, Satoru; Howley, Peter M; Sakai, Hiroyuki

2006-04-01

73

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

International Nuclear Information System (INIS)

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

74

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

Science.gov (United States)

A picosecond pulsed electric field (psPEF) is a localized physical therapy for tumors that has been developed in recent years, and that may in the future be utilized as a targeted non?invasive treatment. However, there are limited studies regarding the biological effects of psPEF on cells. Electric field amplitude and pulse number are the main parameters of psPEF that influence its biological effects. In this study, we exposed HeLa cells to a psPEF with a variety of electric field amplitudes, from 100 to 600 kV/cm, and various pulse numbers, from 1,000 to 3,000. An MTT assay was used to detect the growth inhibition, while flow cytometry was used to determine the occurrence of apoptosis and the cell cycle of the HeLa cells following treatment. The morphological changes during cell apoptosis were observed using transmission electron microscopy (TEM). The results demonstrated that the cell growth inhibition rate gradually increased, in correlation with the increasing electric field amplitude and pulse number, and achieved a plateau of maximum cell inhibition 12 h following the pulses. In addition, typical characteristics of HeLa cell apoptosis in the experimental groups were observed by TEM. The results demonstrated that the rate of apoptosis in the experimental groups was significantly elevated in comparison with the untreated group. In the treatment groups, the rate of apoptosis was greater in the higher amplitude groups than in the lower amplitude groups. The same results were obtained when the variable was the pulse number. Flow cytometric analysis indicated that the cell cycle of the HeLa cells was arrested at the G2/M phase following psPEF treatment. Overall, our results indicated that psPEF inhibited cell proliferation and induced cell apoptosis, and that these effects occurred in a dose-dependent manner. In addition, the results demonstrated that the growth of the HeLa cells was arrested at the G2/M phase following treatment. This study may provide a foundation for further in vivo experiments, and for the potential clinical application of psPEF in the treatment of cervical cancer. PMID:23589101

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

2013-06-01

75

Ascorbic acid: effects on ricin intoxicated HeLa cells  

International Nuclear Information System (INIS)

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

76

Global expression profile of telomerase-associated genes in HeLa cells.  

Science.gov (United States)

Telomerase is a specialized nucleoprotein enzyme complex that maintains the telomere length. The telomerase reverse transcriptase (TERT) is the catalytically active component of the telomerase complex. In humans, the protein component (hTERT) and RNA component (hTR) are found to differentially express in cancer cells. In contrast to differentiated cells, most of the cancer cells overexpress hTERT, which is needed to maintain the proliferative potential of cells. The overexpression of telomerase is not proportionate to telomere length in cancer cells, suggesting that the immortalizing phenotype can be mediated through other factors in addition to telomere length. To investigate the role of hTERT in immortalizing process, loss of gene function studies were carried out. Short interfering RNA (siRNA) and short hairpin RNA (shRNA) against hTERT showed the reduction of hTERT transcript, reduction of telomerase activity and alteration of gene expression in HeLa cells. The molecular basis of proliferative capacity of hTERT was investigated by gene expression microarray. Analysis of microarray data for HeLa cells following siRNA and shRNA mediated knockdown of hTERT showed that 80 genes were upregulated and 73 genes downregulated. Out of these, 37 genes are known to be involved in cancer. Further analyses of previously known genes involved in cancer like KLF4, FGF2, IRF-9 and PLAU by Real Time PCR showed their upregulation. We are documenting for the first time the effect of knocking down hTERT on expression of KLF4 and FGF2. Interestingly, it has been earlier reported that KLF4 and FGF2 up-regulate the expression of hTERT in cancer cells. This suggests that hTERT may be subject to its own auto-regulatory effects. PMID:24929127

Varshney, Akhil; Ramakrishnan, Suresh K; Sharma, Amod; Santosh, Baby; Bala, Jyoti; Yadava, Pramod K; Jaiswal, Rishi Kumar

2014-09-01

77

Cytotoxicity of ICD-85 NPs on Human Cervical Carcinoma HeLa Cells through Caspase-8 Mediated Pathway  

OpenAIRE

The biological application of nanoparticles (NPs) is a rapidly developing area of nanotechnology that raises new possibilities in the treatment of human cancers. The cytotoxicity was evaluated by MTT and LDH assays. The apoptotic effect of free ICD-85 and ICD-85 NPs on HeLa cells was assessed using caspase-8 colorimetric assay. The MTT assay showed that ICD-85 NPs could enhance the in-vitro cytotoxicity against HeLa cells compared to the free ICD-85. The IC50 value at 72 h was reduced from 25...

Moradhaseli, Saeed; Zare Mirakabadi, Abbas; Sarzaeem, Ali; Kamalzadeh, Morteza; Haji Hosseini, Reza

2013-01-01

78

Julibroside J8-induced HeLa cell apoptosis through caspase pathway.  

Science.gov (United States)

The julibroside J8 was isolated from the Albizia julibrissin and evaluated for antiproliferatived on six cancer cell lines (BGC-823, Bel-7402, HeLa, PC-3MIE8, MDA-MB-435 and LH-60) in vitro. Julibroside J8 at 100 microg mL- 1 (46.08 micromol.L- 1) significantly inhibited growth in the first three cell lines. In addition, in HeLa cells typical apoptotic changes in morphology were observed, and further, nuclear damage was observed by Giemsa staining and DNA fragmentation was exhibited. Effects of julibrosideJ8 on induction of DNA fragmentation, caspase-3 activation and downregulation of ICAD expression were effectively inhibited by a caspase-3 inhibitor, z-DEVD-fmk. In addition, apoptosis induced with julibroside J8 was associated with an increase in expression of the apoptosis inducer Bax, and a significant reduction in expression of the apoptosis suppressor Bcl-2 in mitochondria. These results suggest that julibroside J8 induces HeLa death through caspase pathway. PMID:16864463

Zheng, L; Zheng, J; Wu, L-J; Zhao, Y-Y

2006-01-01

79

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

Energy Technology Data Exchange (ETDEWEB)

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.

Yao, Qing; Xu, Hui; Zhang, Qian-Qian; Zhou, Hui [Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory for Biocontrol, Sun Yat-Sen University, Guangzhou 510275 (China); Qu, Liang-Hu, E-mail: lssqlh@mail.sysu.edu.cn [Key Laboratory of Gene Engineering of the Ministry of Education, State Key Laboratory for Biocontrol, Sun Yat-Sen University, Guangzhou 510275 (China)

2009-10-23

80

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

International Nuclear Information System (INIS)

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

81

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

Directory of Open Access Journals (Sweden)

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

Fariba Samani

2014-04-01

82

Plasmid-associated adherence of Shigella flexneri in a HeLa cell model.  

OpenAIRE

The initial interaction of Shigella flexneri with HeLa cells was studied at 4 degrees C, a temperature that inhibits parasite-directed endocytosis. It was found that invasive strains were 10-fold more adherent to HeLa cells than were isogenic, noninvasive strains which had lost a 140-megadalton plasmid. Adherent strains were also more hydrophobic than were nonadherent strains.

Pa?l, T.; Hale, T. L.

1989-01-01

83

The genomic and transcriptomic landscape of a HeLa cell line.  

Science.gov (United States)

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

84

Terbium doped SnO2 nanoparticles as white emitters and SnO2:5Tb/Fe3O4 magnetic luminescent nanohybrids for hyperthermia application and biocompatibility with HeLa cancer cells.  

Science.gov (United States)

SnO2:5Tb (SnO2 doped with 5 at% Tb(3+)) nanoparticles were synthesised by a polyol method and their luminescence properties at different annealing temperatures were studied. Characterization of nanomaterials was done by X-ray diffraction (XRD), Fourier transformation infrared spectroscopy (FTIR), transmission electron microscopy (TEM) and vibrating sample magnetometry (VSM). XRD studies indicate that the prepared nanoparticles were of tetragonal structures. Upon Tb(3+) ion incorporation into SnO2, Sn(4+) changes to Sn(2+) and, on annealing again at higher temperature, Sn(2+) changes to Sn(4+). The prepared nanoparticles were spherical in shape. Sn-O vibrations were found from the FTIR studies. In photoluminescence studies, the intensity of the emission peaks of Tb(3+) ions increases with the increase of annealing temperature, and emission spectra lie in the region of white emission in the CIE diagram. CCT calculations show that the SnO2:5Tb emission lies in cold white emission. Quantum yields up to 38% can be obtained for 900 °C annealed samples. SnO2:5Tb nanoparticles were well incorporated into the PVA polymer and such a material incorporated into the polymer can be used for display devices. The SnO2:5Tb/Fe3O4 nanohybrid was prepared and investigated for hyperthermia applications at different concentrations of the nanohybrid. This achieves a hyperthermia temperature (42 °C) under an AC magnetic field. The hybrid nanomaterial SnO2:5Tb/Fe3O4 was found to exhibit biocompatibility with HeLa cells (human cervical cancer cells) at concentrations up to 74% for 100 ?g L(-1). Also, this nanohybrid shows green emission and thus it will be helpful in tracing magnetic nanoparticles through optical imaging in vivo and in vitro application. PMID:25747103

Singh, Laishram Priyobarta; Singh, Ningthoujam Premananda; Srivastava, Sri Krishna

2015-03-24

85

HeLa cell identification by analysis of ribosomal DNA segment patterns generated by endonuclease restriction.  

OpenAIRE

Restriction endonuclease analysis of HeLa cells and cells in which origins have been questioned provides evidence in favor of a HeLa cell origin for the questioned cells. Digestion of cellular human DNA reveals a variable ribosomal DNA (rDNA) fragment that is present in up to four discrete sizes. Cell lines of known and suspected HeLa origin contain only two size variants. This pattern of variability serves to distinguish HeLa-derived cells from others. Despite repeated passage and divergence...

Schmickel, R. D.; Waterson, J. R.; Knoller, M.; Szura, L. L.; Wilson, G. N.

1980-01-01

86

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)

87

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

OpenAIRE

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

88

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

International Nuclear Information System (INIS)

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)

89

A lipidomics investigation of the induced hypoxia stress on HeLa cells by using MS and NMR techniques.  

Science.gov (United States)

Induced hypoxia stress on cervical cancer derived cells (HeLa cells) leads to significant changes in their membrane lipid profiles. The lipidome of HeLa cells was characterized by a joint approach wherein liquid chromatography-mass spectrometry (LC-MS) analysis was followed by high resolution NMR measurements. Multivariate data analysis showed apparent separation between control and hypoxia-treated HeLa cells and thus demonstrated hypoxia effects on lipid metabolism. The most striking finding was that hypoxia stimulation significantly reduced the total amount of cellular phosphoinositols (PI) but caused a prominent increase in the amount of lyso phosphocholines (lyso-PC) and lyso phosphoethanolamines (lyso-PE). The observed decrease of PI amount under hypoxic conditions is probably due to the accumulation of cellular myo-inositol, which is known to play a critical role in de novo synthesis of PI. Moreover, our study suggests that polyunsaturated phospholipid species are stronger biomarkers for discriminating the effect of hypoxia treatment. The evaluation of changes in the average unsaturation index (UI) of the membrane lipids acyl chains reveals that UI slightly increases in several lipid classes, thus affecting membrane fluidity and further membrane-dependent functions. The plausible mechanisms by which HeLa cells adapt to hypoxia conditions are also briefly reported. PMID:24496110

Yu, Yang; Vidalino, Laura; Anesi, Andrea; Macchi, Paolo; Guella, Graziano

2014-04-01

90

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

Science.gov (United States)

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

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

2014-08-01

91

A novel L1 retrotransposon marker for HeLa cell line identification  

OpenAIRE

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

92

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

Directory of Open Access Journals (Sweden)

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

Wahyu Widowati

2013-06-01

93

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

94

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

Energy Technology Data Exchange (ETDEWEB)

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

95

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

Directory of Open Access Journals (Sweden)

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

96

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

Science.gov (United States)

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

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

2015-01-01

97

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

Directory of Open Access Journals (Sweden)

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

Yuxin Liu

2012-06-01

98

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

OpenAIRE

There is a lack of biological studies using selenium and tellurium compounds, specially concern with cancer therapy. The aim of this study is to evaluate the cytotoxicity action of nine ?-chalcogen-substituted Michael-aldol-type adducts on HeLa and RKO cancer cell lines. The cytotoxic effect was assessed by MTT assay and was performed in HeLa and RKO cell lines cultured in RPMI-1640 medium in (95% O2+5% CO2) at 37°C. The IC50 values demonstrated that all compounds presented cytotoxic ...

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

2013-01-01

99

A novel bispidinone analog induces S?phase cell cycle arrest and apoptosis in HeLa human cervical carcinoma cells.  

Science.gov (United States)

2,4,6,8-(3)-Tetranitrophenyl-3,7-diazabicyclo[3.3.1]nonan-9-one (B16), a bispidinone analog, was synthesized to investigate its effects on cell viability, the cell cycle, and apoptotic pathways in HeLa human cervical cancer cells. B16 decreased the percentage of viable cells in WST-8 assays, and morphological changes associated with apoptotic cell death were observed, including cell shrinkage and disruption. Annexin V-FITC/PI dual staining assays showed that B16 significantly increased the early apoptosis of HeLa cells after 24 h of treatment. Moreover, DNA content analysis and [3H]-thymidine incorporation assays showed that B16 induced S-phase cell cycle arrest and inhibited DNA replication after 24 h of treatment. Following treatment with 25 µM of B16, an increase in reactive oxygen species and a decrease in mitochondrial membrane potential were observed by flow cytometry. In addition, the expression levels of caspase cascade and Bcl-2 family proteins determined by western blotting suggested that the induction of apoptosis by B16 was associated with a caspase- and mitochondrial-dependent pathway in HeLa cells. In conclusion, B16 induced early apoptosis and S-phase cell cycle arrest in HeLa cells via a caspase- and mitochondrial?dependent pathway. PMID:25592775

Yi, Xu; Zhang, Xin; Jeong, Hyunjin; Shin, Yu Mi; Park, Dong Ho; You, Song; Kim, Dong-Kyoo

2015-03-01

100

Effect of tunicamycin on cisplatin induced apoptosis of HeLa cells  

Directory of Open Access Journals (Sweden)

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

101

Putative mechanisms of antitumor activity of cyano-substituted heteroaryles in HeLa cells.  

OpenAIRE

Six recently synthesized cyano-substituted heteroaryles, which do not bind to DNA but are highly cytotoxic against the human tumor cell line HeLa, were analyzed for their antitumor mechanisms of action (MOA). They did not interfere with the expression of human papillomavirus oncogenes integrated in the HeLa cell genome, but they did induce strong G1 arrest and result in the activation of caspase-3 and apoptosis. A computational analysis was performed that compared the antiproliferative activi...

Lembo, David; Donalisio, Manuela

2010-01-01

102

Trypanosoma cruzi trypomastigotes induce cytoskeleton modifications during HeLa cell invasion  

OpenAIRE

It has been recently shown that Trypanosoma cruzi trypomastigotes subvert a constitutive membrane repair mechanism to invade HeLa cells. Using a membrane extraction protocol and high-resolution microscopy, the HeLa cytoskeleton and T. cruzi parasites were imaged during the invasion process after 15 min and 45 min. Parasites were initially found under cells and were later observed in the cytoplasm. At later stages, parasite-driven protrusions with parallel filaments were observed, with trypoma...

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

2011-01-01

103

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

Directory of Open Access Journals (Sweden)

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

Alimohammad Asgharian

2013-01-01

104

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

Science.gov (United States)

One of the most significant steps in gene expression studies is transferring genes into cell cultures. Despite there are different methods for gene delivery such as viral and non-viral producers, some cationic lipid reagents have recently developed to transfect into mam- malian cell lines. The main aim of this study was optimizing and improving lipocomplex based transient transfection procedures into HeLa cell line which is being used widely as a typical cell in biological studies. This study was an experimental research. In this work, pCMV ?-Gal DNA plasmid was used as a reporter DNA for determining the rate of gene transfection into HeLa cells. To accomplish the highest gene delivery into HeLa cells, optimizing experiments were car- ried out in different volumes of FuGENE-HD, Lipofectamine(TM)2000 and X-tremeGENE. Also, we investigated tranasfection efficiency in presence of various cell densities of HeLa cells. Then, transfection efficiency and cell toxicity were measured by beta gal staining and trypan blue methods, respectively. Using FuGENE-HD in volume of 4µl along with 10(5) HeLa cells, transfection efficiency was higher (43.66 ± 1.52%) in comparison with the cationic lipids Lipofectamine(TM)2000 and X-tremeGENE. In addition, the rate of cell toxicity in presence of FuGENE-HD was less than 5%. In summary, the cationic lipid FuGENE-HD indicates a suitable potential to transfer DNA into HeLa cells and it can be an efficient reagent for gene delivery for HeLa cells in vitro. Moreover, it is worth designing and optimizing gene transfer experiments for other cell lines with FuGENE-HD due to its low toxicity and high efficiency. PMID:24381863

Asgharian, Alimohammad; Banan, Mehdi; Najmabadi, Hossein

2014-01-01

105

Methanolic Extracts from Brown Seaweeds Dictyota cilliolata and Dictyota menstrualis Induce Apoptosis in Human Cervical Adenocarcinoma HeLa Cells.  

Science.gov (United States)

Carcinoma of the uterine cervix is the second most common female tumor worldwide, surpassed only by breast cancer. Natural products from seaweeds evidencing apoptotic activity have attracted a great deal of attention as new leads for alternative and complementary preventive or therapeutic anticancer agents. Here, methanol extracts from 13 species of tropical seaweeds (Rhodophytas, Phaeophyta and Chlorophyta) collected from the Northeast of Brazil were assessed as apoptosis-inducing agents on human cervical adenocarcinoma (HeLa). All extracts showed different levels of cytotoxicity against HeLa cells; the most potent were obtained from the brown alga Dictyota cilliolata (MEDC) and Dictyota menstrualis (MEDM). In addition, MEDC and MEDM also inhibits SiHa (cervix carcinoma) cell proliferation. Studies with these two extracts using flow cytometry and fluorescence microscopy showed that HeLa cells exposed to MEDM and MEDC exhibit morphological and biochemical changes that characterize apoptosis as shown by loss of cell viability, chromatin condensation, phosphatidylserine externalization, and sub-G1 cell cycle phase accumulation, also MEDC induces cell cycle arrest in cell cycle phase S. Moreover, the activation of caspases 3 and 9 by these extracts suggests a mitochondria-dependent apoptosis route. However, other routes cannot be ruled out. Together, these results point out the methanol extracts of the brown algae D. mentrualis and D. cilliolata as potential sources of molecules with antitumor activity. PMID:25871374

Gomes, Dayanne Lopes; Telles, Cinthia Beatrice Silva; Costa, Mariana Santana Santos Pereira; Almeida-Lima, Jailma; Costa, Leandro Silva; Keesen, Tatjana Souza Lima; Rocha, Hugo Alexandre Oliveira

2015-01-01

106

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)

107

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

Science.gov (United States)

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

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

1993-03-25

108

Photodynamic damage study of HeLa cell line using ALA  

Science.gov (United States)

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

109

Heterofucan from Sargassum filipendula Induces Apoptosis in HeLa Cells  

Directory of Open Access Journals (Sweden)

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

110

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  

Directory of Open Access Journals (Sweden)

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

111

The effect of knocking-down nucleostemin gene expression on the in vitro proliferation and in vivo tumorigenesis of HeLa cells.  

Science.gov (United States)

Previous studies have demonstrated nucleostemin (NS) expressed in undifferentiated cells (e.g. embryonic stem cells and myeloid stem cells) and various cancer cell lines, but didn't express in differentiated adult tissues. In this study, we examined the NS expression in several cancer cell lines and some normal human tissues. Moreover, we used RNAi techniques to knock down the NS gene expression in Hela cells. As a result, we found that all these detected cancer cell lines in our program had high levels of NS expression, which indicated NS played a really important role in the self-renewal of these cancer cells. At the same time, NS expressed in human placenta tissue also at a high level, which may be due to the existence of a mass of placenta stem cells. Only a small quantity of NS expression could be found in normal human muscle tissue due to the existence of myoblasts. And for the first time our findings demonstrated that if NS expression of Hela cells was knocked down, more Hela cells couldn't complete the DNA synthesis to pass through S phase. So the percent of G0/G1 phase increased with the decrease of S phase and cell proliferation rate and in vivo tumorigenic capacity decreased obviously. Our work would help shed light on the self-renewal and malignant growth of cancer cells. PMID:15595646

Sijin, L; Ziwei, C; Yajun, L; Meiyu, D; Hongwei, Z; Guofa, H; Siguo, L; Hong, G; Zhihong, Z; Xiaolei, L; Yingyun, W; Yan, X; Weide, L

2004-09-01

112

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

International Nuclear Information System (INIS)

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

113

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

114

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)

115

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

Science.gov (United States)

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

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

2009-01-01

116

Correlation in HeLa cells of anchorage-independent growth and synthesis of the glycoprotein hormone alpha-subunit.  

Science.gov (United States)

In addition to its normal synthesis in pituitary and placenta for production of the four glycoprotein hormones (GPH), the free alpha-subunit is produced by a variety of tumors and tumor-derived cell lines. It has long been used clinically as a tumor marker, but the question remains as to whether ectopic production of GPH alpha represents the random and chance activation of the hormone gene during cancer development or whether GPH alpha may contribute directly or indirectly to the biology of the tumor. One characteristic of tumorigenic cells in culture is their ability to proliferate in an anchorage-independent way. Data are presented in the following paper to show that the cloning efficiency of HeLa cervical carcinoma cells in soft agar is directly correlated with their production of the GPH alpha-subunit. HeLa variants that differ over 400-fold in their production of GPH alpha similarly exhibited a marked difference in their ability to form colonies in 0.3% noble agar. When HeLa SR3 cells (a variant that produces GPH alpha at high levels) was stably transfected with a vector producing the antisense strand of GPH alpha cDNA, the synthesis of the GPH alpha-subunit was reduced in these cells as was their cloning efficiency in soft agar. Similarly, when HeLa A5F cells (a variant producing little or no GPH alpha) were stably transfected with an alpha-subunit expression vector, the production of GPH alpha was increased significantly in concert with an increased ability to form colonies in 0.3% noble agar. Somatic cell hybrids between HeLa SR3 and HeLa A5F exhibited intermediate levels of GPH alpha gene expression and colony formation in soft agar compared to the parental strains. These data suggest that some parameters of the tumorigenic phenotype, such as anchorage-independent growth, are responsive to, or are dependent upon, the production of free alpha-subunit. PMID:9144551

Cox, G S

1997-04-17

117

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

Energy Technology Data Exchange (ETDEWEB)

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

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

2013-09-15

118

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

International Nuclear Information System (INIS)

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

119

Comparative susceptibilities of human embryonic fibroblasts and HeLa cells for isolation of human rhinoviruses.  

OpenAIRE

The recovery of human rhinovirus (HRV) from nasal washings and nasal and pharyngeal swabs from volunteers with naturally acquired colds was compared in different cell types. Human embryonic lung fibroblast (HELF) strain WI-38 (sensitivity, 61 to 84%) and HeLa-I, an HRV-susceptible HeLa clone (sensitivity, 86 to 94%), were the most sensitive cell types used. HELF-WI-38 cells showed a cytopathic effect earlier than the other cells used, and the different strains of HRV-susceptible HeLa cells va...

Arruda, E.; Crump, C. E.; Rollins, B. S.; Ohlin, A.; Hayden, F. G.

1996-01-01

120

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

Science.gov (United States)

Overexpression of Bcl-2 has been recognized in various malignancies. Recently, HA14-1, a Bcl-2 antagonist, has been identified for its anti-apoptotic effect. However, mode of action of HA14-1 still remains to be elucidated. In this study, we examined HA14-1 binding efficiency with receptor proteins through molecular docking. Cell viability using HeLa cells was evaluated through MTT assay after exposure to different concentration of HA14-1. Moreover, after HA14-1 exposure, expressions of tumor suppressor protein (p53), BH3-only protein (Puma) and apoptosis-associated proteins were analyzed by Western blotting. From the results, it was found that HA14-1 occupied all three domains; BH1, BH2, and BH3 within the hydrophobic pocket of Bcl-2. However, HA14-1 occupied only BH1 and BH3 of Bcl-xl, conversely, no such stable bond was observed for Bax and Bak. ARG107 and TYR101 were the amino acids involved in the binding of HA14-1 to Bcl-2 and Bcl-xl, respectively. Additionally, decrease in Bcl-2 and Bcl-xl expression along with increase in p53 and Puma expression after exposure to HA14-1 was observed. The results suggested p53 pathway to be the probable mechanism of action for the induction of apoptosis in HeLa cell by downregulating the effect of anti-apoptotic proteins suggesting that HA14-1 may provide therapeutic potential for the treatment of human cervical cancer. PMID:24118733

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

2014-03-01

121

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

OpenAIRE

Bacillus Calmette-Guerin (BCG) immunotherapy is established as an effective adjuvant intravesical treatment for non-muscle invasive bladder cancer. BCG is also effective in the treatment of Condylomata acuminata caused by low-risk human papilloma virus (HPV). The aim of this study was to determine the efficacy of BCG for the treatment of cervical cancer or HPV high-risk infections. BCG-activated killer (BAK) cells were incubated with a high-risk HPV18-infected cervical cancer cell line, HeLa....

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

2012-01-01

122

A novel L1 retrotransposon marker for HeLa cell line identification.  

Science.gov (United States)

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 development of a HeLa-specific DNA diagnostic test: a single duplex detection PCR assay targeting an L1 retrotransposon insertion. All HeLa clones from a geographically diverse panel were positive by this assay, and the particular L1 insertion we identified appears to be unique to the HeLa cell line. The assay can detect very low levels of HeLa contamination (<1%), and can be performed on un-purified cell pellets, allowing rapid routine screening. PMID:19450234

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

2009-04-01

123

FV peptide induces apoptosis in HEp 2 and HeLa cells: an insight into the mechanism of induction  

OpenAIRE

Abstract The present study is an attempt to evaluate the antiproliferative potential of peptide (7.6 kDa) from lionfish (Pterios volitans) venom on cultured HEp2 and HeLa cells. Different dose of purified peptide (1, 2 and 4 ?g/ml) at different time points (12, 24 and 36 hrs) were tested for antiproliferative index of the peptide. Among them, 2 ?g/ml at 24 hrs was found to effectively inhibit cancer cell growth in vitro and did not cause any adverse effect on normal human lymphocyte...

Sri Balasubashini M; Karthigayan S; St, Somasundaram; Balasubramanian T; Rukkumani R; Menon Venugopal P

2006-01-01

124

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

Science.gov (United States)

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

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

2009-05-01

125

Curcumin targeting the thioredoxin system elevates oxidative stress in HeLa cells  

Energy Technology Data Exchange (ETDEWEB)

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

126

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

International Nuclear Information System (INIS)

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

127

Caveolin-1 and CDC42 mediated endocytosis of silica-coated iron oxide nanoparticles in HeLa cells  

Science.gov (United States)

Summary Nanomedicine is a rapidly growing field in nanotechnology, which has great potential in the development of new therapies for numerous diseases. For example iron oxide nanoparticles are in clinical use already in the thermotherapy of brain cancer. Although it has been shown, that tumor cells take up these particles in vitro, little is known about the internalization routes. Understanding of the underlying uptake mechanisms would be very useful for faster and precise development of nanoparticles for clinical applications. This study aims at the identification of key proteins, which are crucial for the active uptake of iron oxide nanoparticles by HeLa cells (human cervical cancer) as a model cell line. Cells were transfected with specific siRNAs against Caveolin-1, Dynamin 2, Flotillin-1, Clathrin, PIP5K? and CDC42. Knockdown of Caveolin-1 reduces endocytosis of superparamagnetic iron oxide nanoparticles (SPIONs) and silica-coated iron oxide nanoparticles (SCIONs) between 23 and 41%, depending on the surface characteristics of the nanoparticles and the experimental design. Knockdown of CDC42 showed a 46% decrease of the internalization of PEGylated SPIONs within 24 h incubation time. Knockdown of Dynamin 2, Flotillin-1, Clathrin and PIP5K? caused no or only minor effects. Hence endocytosis in HeLa cells of iron oxide nanoparticles, used in this study, is mainly mediated by Caveolin-1 and CDC42. It is shown here for the first time, which proteins of the endocytotic pathway mediate the endocytosis of silica-coated iron oxide nanoparticles in HeLa cells in vitro. In future studies more experiments should be carried out with different cell lines and other well-defined nanoparticle species to elucidate possible general principles. PMID:25671161

Jordan, Andreas

2015-01-01

128

Dynamic behavior of histone H1 microinjected into HeLa cells  

International Nuclear Information System (INIS)

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

129

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

Science.gov (United States)

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

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

2014-01-24

130

p150 ADAR1 isoform involved in maintenance of HeLa cell proliferation  

Directory of Open Access Journals (Sweden)

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

131

Radiosensitivity of Hela cells in various O2 concentrations and consideration of oxygen effect in radiotherapy  

International Nuclear Information System (INIS)

The aim of this paper is the study of the radiosensitivity of HeLa cells in vitro in various oxygen concentrations and the consideration of the utilization of oxygen effect in radiation therapy, based on the data of HeLa cells and tumor oxygen tension. Survival curves of HeLa cells are found to be exponential as a function of radiation dose and the radiosensitivity is dependent on oxygen tension of culture medium. Relative radiosensitivity decreases remarkably at low level of oxygen, especially under 9 mmHg pO2. The utilization of oxygen effect in radiation may be useful in hyperbaric oxygen inhalation and not useful under local tissue hypoxia induced by tourniquet application. Reoxygenation occurs with shrinkage of tumor after irradiation and this phenomenon will diminish the value of hyperbaric oxygen in radiation therapy. (author)

132

In vitro studies of the toxic effects of silver nanoparticles on HeLa and U937 cells  

Science.gov (United States)

In the last decade, much attention has been paid to studies of the effect of silver nanoparticles (Ag NPs) on tumor cells. Apart from elucidation of the mechanism of NPs’ interaction with mammalian cells, these studies are aimed at discovering new effective antitumor drugs. In this work, we report about the toxic effects of Ag NPs observed on two types of tumor cells: HeLa (adhesive cells) and U937 (suspension cells). The Ag NPs were obtained by an original method of biochemical synthesis. Particle size was 13.2±4.72 nm, and zeta potential was ?61.9±3.2 mV. The toxicity of Ag NPs in the concentration range 0.5–8.0 ?g Ag/mL was determined by means of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and cytofluorometry after 4 and 24 hours’ incubation. It was found that Ag NPs had high toxicity toward both cell types. The minimal concentrations where a toxicity effect was registered (toxicity thresholds) lied in the range 0.5–2.0 ?g Ag/mL. In parallel with the Ag NP solution, cells were incubated with water solutions of the NP stabilizer (aerosol-OT) and Ag+ ions (as silver nitrate). It was shown that aerosol-OT had no effect on the viability on HeLa cells, but was moderately toxic toward U937, though less dangerous for these cells than Ag NPs. With Ag+ ions, for HeLa no toxic effect was observed, while for U937 they were as toxic as the Ag NPs. The data obtained indicate that Ag NPs as used in this study may prove to be useful for the creation of medicines for cancer therapy. PMID:25784794

Kaba, Said I; Egorova, Elena M

2015-01-01

133

Trypanosoma cruzi trypomastigotes induce cytoskeleton modifications during HeLa cell invasion  

Directory of Open Access Journals (Sweden)

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

Maria Cecília Fernandes

2011-12-01

134

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

OpenAIRE

The aim of this study was to observe the cytotoxicity and apoptotic effects of nickel oxide nanoparticles on human cervix epithelioid carcinoma cell line (HeLa). Nickel oxide precursors were synthesized by an nickel sulphate-excess urea reaction in boiling aqueous solution. The synthesized NiO nanoparticles (<200 nm) were investigated by X-ray diffraction analysis and transmission electron microscopy techniques. For cytotoxicity experiments, HeLa cells were incubated in 50-500 Î?g/mL NiO...

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

2010-01-01

135

Inactivation of mitotic factors by ultraviolet irradiation of HeLa cells in mitosis  

International Nuclear Information System (INIS)

Extracts from mitotic HeLa cells, when injected into fully grown Zenopus laevis oocytes, exhibit maturation-promoting activity (MPA) indicated by germinal vesicle breakdown (GVBD) and chromosome condensation. The MPA of mitotic cell extracts is neutralized by the inhibitors of mitotic factors (IMF) in HeLa cells. The activity of the IMF coincides with chromosome condensation. A study was performed to investigate whether chromosome decondensation and the activation of IMF were related. The activity of IMF was measured in N2O-blocked mitotic HeLa cells, in which chromosome decondensation was induced by exposure to ultraviolet light, and subsequent incubation in medium containing inhibitors of DNA synthesis. U.v. irradiation activated IMF in mitotic HeLa cells but not in cell extracts. In contrast, much less activation of IMF was seen at very high doses of X-irradiation. The IMF seemed to inactivate the mitotic factors directly by forming a complex that precipitated on heating at 600C. Mg2+ or polyamine agents known to promote chromatin condensation partially restored the MPA of the u.v.-irradiated mitotic cell extracts. These results tend to support the conclusion that the IMF play a role in the decondensation of chromosomes. (author)

136

Anticancer activity and mechanism of juglone on human cervical carcinoma HeLa cells.  

Science.gov (United States)

Induction of apoptosis in tumor cells has become the major focus of anti-tumor therapeutics development. Juglone, a major chemical constituent of Juglans mandshurica Maxim, possesses several bioactivities, including anti-tumor. In the present study, HeLa cells were incubated with juglone at various concentrations. The proliferation inhibition of juglone on HeLa cells was tested by the MTT assay. Occurrence of apoptosis was detected by Hoechst 33258 staining, flow cytometry, and transmission electron microscopy. The expression of apoptotic-related proteins was examined by Western blot. The results showed that juglone inhibits the growth of HeLa cells in dose-dependent manner. Topical morphological changes of apoptotic body formation after juglone treatment were observed. The percentages of early apoptosis of Annexin V-FITC were 5.23%, 7.95%, 10.69%, and 20.92% with the concentrations of juglone (12.5, 25, 50, and 100 µmol/L), respectively. After cells were treated with juglone at the different dose for 24 h, the expression of Bcl-2 was significantly down-regulated and the expression of Bax was significantly up-regulated compared with the control. These events paralleled with activation of caspase-9, -8, -3, and PARP cleavage. The results suggest that juglone may be effective for the treatment of HeLa cells. PMID:23181283

Zhang, Wei; Liu, Anheng; Li, Yan; Zhao, Xingyu; Lv, Shijie; Zhu, Wenhe; Jin, Ying

2012-11-01

137

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

International Nuclear Information System (INIS)

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

138

Interphase death of HeLa Zh-63 cells induced by high doses of ?-radiation  

International Nuclear Information System (INIS)

Interphase death of HeLa cells has been assessed by the ability of cells to be stained with trypan blue after ?-irradiation with doses from 50 to 2000 krad. A quantitative relationship between average life span of cells and radiation dose has been shown. A decrease of the postirradiation temperature and an increase of the serum content in the growth medium before irradiation have been found to delay the interphase death. Adhesive properties of cells are damaged immediately after exposure

139

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

OpenAIRE

One of the most significant steps in gene expression studies is transferring genes into cell cultures. Despite there are different methods for gene delivery such as viral and non-viral producers, some cationic lipid reagents have recently developed to transfect into mammalian cell lines. The main aim of this study was optimizing and improving lipocomplex based transient transfection procedures into HeLa cell line which is being used widely as a typical cell in biological studies.This study wa...

Alimohammad Asgharian; Mehdi Banan; Hossein Najmabadi

2013-01-01

140

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

141

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

Directory of Open Access Journals (Sweden)

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

Julie Kirkegaard

2014-08-01

142

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

Directory of Open Access Journals (Sweden)

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

143

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

Directory of Open Access Journals (Sweden)

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

144

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

Science.gov (United States)

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

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

1991-12-01

145

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

146

Promotion of cell proliferation and inhibition of ADCC by cancerous immunoglobulin expressed in cancer cell lines  

OpenAIRE

To explore the significance of cancerous immunoglobulin (Ig) in cancer cell growth, HeLa cervical cancer cells were stably transfected with small interfering RNA (siRNA) that specifically, efficiently and consistently silences the expression of heavy chain genes of all immunoglobulin isotypes. This stable cell line was used to examine cell viability, colony formation and tumor growth in athymic nude mice. The results of these experiments indicated that siRNA-mediated knockdown of cancerous Ig...

Li, Ming; Zheng, Hui; Duan, Zhi; Liu, Haidan; Hu, Duosha; Bode, Ann; Dong, Zigang; Cao, Ya

2011-01-01

147

Vaccinia virus-induced changes in [Na+] and [K+] in HeLa cells.  

Science.gov (United States)

A flame photometric technique is described for determining average values for intracellular [Na+] and [K+] in HeLa cells. Ion measurements were made on unwashed cells disrupted ultrasonically in the presence of residual medium; corrections for the latter were made by measurement of extracellular volume in cell plus medium preparations using 125I-labelled polyvinylpyrrolidone (PVP) as the marker in an isotopic dilution technique. Accurate measurement of the volume occupied by the cells was critical and required a concentration step. This was achieved by concentrating cell suspensions in a microhaematocrit centrifuge using calibrated capillary tubes. Most reliable values were obtained in our system using HeLa S3 (suspension) cells grown as monolayers, which were removed by EDTA and held in suspension for a minimum of 2 h. Uninfected HeLa cells had values of 20 to 30 and 110 to 120 mM for Na+ and K+ respectively. At 13 h after inoculation with vaccinia virus, a dramatic reversal in [Na+] and [K+] occurred, but throughout the infection cycle the total [Na+ + K+] varied little. The significance of these data is discussed in relation to theories of virus-induced modulation of protein synthesis in infected cells and in cell-free systems. PMID:7130948

Norrie, D H; Wolstenholme, J; Howcroft, H; Stephen, J

1982-09-01

148

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

149

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

International Nuclear Information System (INIS)

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

150

Stable gene amplification and overexpression of sodium- and potassium-activated ATPase in HeLa cells.  

OpenAIRE

Cell lines stably resistant to ouabain were isolated from an unstably resistant HeLa line after growth in nonselective medium. Stable resistant lines bound ouabain at levels 10-fold higher than did HeLa cells and at similar levels to those bound by the unstable C+ line previously described (J. F. Ash, R. M. Fineman, T. Kalka, M. Morgan, and B. Wire, J. Cell Biol. 99: 971-983). Expression and synthesis of the Na+, K+ -ATPase alpha chain showed a similar amplification over that for HeLa cells b...

Pauw, P. G.; Johnson, M. D.; Moore, P.; Morgan, M.; Fineman, R. M.; Kalka, T.; Ash, J. F.

1986-01-01

151

Alsterpaullone, a Cyclin-Dependent Kinase Inhibitor, Mediated Toxicity in HeLa Cells through Apoptosis-Inducing Effect  

OpenAIRE

Alsterpaullone, a small molecule cyclin-dependent kinase (CDK) inhibitor, regulates the cell cycle progression. Beyond death-inducing properties, we identified the effect of alsterpaullone on cycle procedure and apoptosis of HeLa cell. It was found that alsterpaullone inhibited HeLa cells in a time-dependent (0–72?h) and dose-dependent (0–30??M) manner. In the presence of alsterpaullone, HeLa cells were arrested in G2/M prior to undergoing apoptosis via a mechanism that is involved i...

Chunying Cui; Yuji Wang; Yaonan Wang; Ming Zhao; Shiqi Peng

2013-01-01

152

Cell damage resulting from the labeling of rat lymphocytes and HeLaS3 cells with In-111 oxine  

Energy Technology Data Exchange (ETDEWEB)

Rat thoracic-duct lymphocytes and HeLa S3 cells were labeled in vitro with different amounts of indium-111 oxine. The labeled rat lymphocytes were tested for their ability to recirculate normally in syngeneic rats; the labeled HeLa S3 cells for their ability to divide to form colonies in tissue culture. Both cell types behaved normally by these criteria when labeled with small amounts of indium-111 oxine but at higher doses were obviously damaged. Evidence was obtained for the HeLa S3 cells that this damage was primarily radiation-induced. These findings may impose limitations on the use of In-111 oxine as a cell label for clinical purposes.

Chisholm, P.M.; Danpure, H.J.; Healey, G.; Osman, S.

1979-12-01

153

Cell damage resulting from the labeling of rat lymphocytes and HeLaS3 cells with In-111 oxine  

International Nuclear Information System (INIS)

Rat thoracic-duct lymphocytes and HeLa S3 cells were labeled in vitro with different amounts of indium-111 oxine. The labeled rat lymphocytes were tested for their ability to recirculate normally in syngeneic rats; the labeled HeLa S3 cells for their ability to divide to form colonies in tissue culture. Both cell types behaved normally by these criteria when labeled with small amounts of indium-111 oxine but at higher doses were obviously damaged. Evidence was obtained for the HeLa S3 cells that this damage was primarily radiation-induced. These findings may impose limitations on the use of In-111 oxine as a cell label for clinical purposes

154

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

155

Single-walled carbon nanotube interactions with HeLa cells  

OpenAIRE

Abstract This work concerns exposing cultured human epithelial-like HeLa cells to single-walled carbon nanotubes (SWNTs) dispersed in cell culture media supplemented with serum. First, the as-received CoMoCAT SWNT-containing powder was characterized using scanning electron microscopy and thermal gravimetric analyses. Characterizations of the purified dispersions, termed DM-SWNTs, involved atomic force microscopy, inductively coupled plasma – mass spectrometry, and absorption and Raman spect...

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

2007-01-01

156

Active RNA polymerase I is fixed within the nucleus of HeLa cells.  

OpenAIRE

We have investigated whether active RNA polymerase I, the enzyme responsible for transcribing ribosomal RNA, is immobilized by attachment to a large subnuclear structure in HeLa cells. As unphysiological salt concentrations induce artifacts, we have used isotonic conditions throughout the preparative and analytic procedures. Cells are encapsulated in agarose microbeads and lysed in Triton and a 'physiological' buffer; then soluble proteins and RNA diffuse out through the agarose pores to leav...

Dickinson, P.; Cook, Pr; Jackson, Da

1990-01-01

157

Establishment of HeLa Cell Mutants Deficient in Sphingolipid-Related Genes Using TALENs  

OpenAIRE

Sphingolipids are essential components in eukaryotes and have various cellular functions. Recent developments in genome-editing technologies have facilitated gene disruption in various organisms and cell lines. We here show the disruption of various sphingolipid metabolic genes in human cervical carcinoma HeLa cells by using transcription activator-like effector nucleases (TALENs). A TALEN pair targeting the human CERT gene (alternative name COL4A3BP) encoding a ceramide transport protein ind...

Yamaji, Toshiyuki; Hanada, Kentaro

2014-01-01

158

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

Directory of Open Access Journals (Sweden)

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

Kezban Ada

2010-04-01

159

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

Directory of Open Access Journals (Sweden)

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

Nisa Tandogan

2011-04-01

160

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

Directory of Open Access Journals (Sweden)

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

I. JURANIC

1999-09-01

161

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.

162

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

International Nuclear Information System (INIS)

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

163

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

Science.gov (United States)

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

Sato, Takehiko; Yokoyama, Mayo; Johkura, Kohei

2011-09-01

164

Alsterpaullone, a Cyclin-Dependent Kinase Inhibitor, Mediated Toxicity in HeLa Cells through Apoptosis-Inducing Effect  

Science.gov (United States)

Alsterpaullone, a small molecule cyclin-dependent kinase (CDK) inhibitor, regulates the cell cycle progression. Beyond death-inducing properties, we identified the effect of alsterpaullone on cycle procedure and apoptosis of HeLa cell. It was found that alsterpaullone inhibited HeLa cells in a time-dependent (0–72?h) and dose-dependent (0–30??M) manner. In the presence of alsterpaullone, HeLa cells were arrested in G2/M prior to undergoing apoptosis via a mechanism that is involved in the regulation of various antiapoptotic genes, DNA-repair, transcription, and cell cycle progression. Compared to controls, alsterpaullone effectively prevented HeLa cells from entering S-phase. These potential therapeutic efficacies could be correlated with the activation of caspase-3. PMID:23577282

Cui, Chunying; Wang, Yuji; Wang, Yaonan; Zhao, Ming; Peng, Shiqi

2013-01-01

165

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

166

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)

167

Surface glycosaminoglycans mediate adherence between HeLa cells and Lactobacillus salivarius Lv72  

OpenAIRE

Abstract Background The adhesion of lactobacilli to the vaginal surface is of paramount importance to develop their probiotic functions. For this reason, the role of HeLa cell surface proteoglycans in the attachment of Lactobacillus salivarius Lv72, a mutualistic strain of vaginal origin, was investigated. Results Incubation of cultures with a variety of glycosaminoglycans (chondroitin sulfate A and C, heparin and heparan sulfate) resulted in marked binding interference. However, no single gl...

Marti?n, Rebeca; Marti?n, Carla; Escobedo, Susana; Sua?rez, Juan E.; Quiro?s, Luis M.

2013-01-01

168

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

OpenAIRE

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 ...

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

2011-01-01

169

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). The result s...

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

2010-01-01

170

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

171

Identification of HPV Integration and Gene Mutation in HeLa Cell Line by Integrated Analysis of RNA-Seq and MS/MS Data.  

Science.gov (United States)

HeLa cell line, which was derived from cervical carcinoma, provides an idea platform to study both the integration of human papillomavirus and the massive mutations occurring on the cancer cell genome. Proteogenomics is a field with the intersection of proteomics and genomics to perform gene annotation and identify gene mutation. In this work, we first identified the SNV/INDEL, structural variation (SV), and virus infection/integration events from RNA-Seq data of HeLa cell line; then, by applying proteogenomics strategy, we were able to detect some of the genomic events with the tandem mass spectrometry (MS/MS) data from the same sample. Furthermore, some of the mutated peptides were experimentally validated using multiple reaction monitoring technology. The integrated analysis of the RNA-Seq and MS/MS data not only renders the discovery of HeLa cell genome variations more credible but also illustrates a practical workflow for protein-coding mutation discovery in cancer-related studies. PMID:25698088

Sun, Han; Chen, Chen; Lian, Baofeng; Zhang, Menghuan; Wang, Xiaojing; Zhang, Bing; Li, Yixue; Yang, Pengyuan; Xie, Lu

2015-04-01

172

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

173

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

174

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

175

Application of microarrays to identify and characterize genes involved in attachment dependence in HeLa cells  

OpenAIRE

The ability to modify cellular properties such as adhesion is of interest in the design and performance of biotechnology-related processes. The current study was undertaken in order to evaluate the effectiveness of modulating cellular adhesion in HeLa cells from a genomics perspective. Using DNA microarrays, differences in gene expression between two phenotypically distinct, anchorage-dependent and anchorage-independent, HeLa cell lines were identified. With the aid of several statistical met...

Jaluria, Pratik; Betenbaugh, Michael; Konstantopoulos, Konstantinos; Frank, Bryan; Shiloach, Joseph

2006-01-01

176

The HeLa cell receptor for enterovirus 70 is decay-accelerating factor (CD55).  

OpenAIRE

Enterovirus 70 (EV70) is a recently emerged human pathogen belonging to the family Picornaviridae. The ability of EV70 to infect a wide variety of nonprimate cell lines in vitro is unique among human enteroviruses. The importance of virus receptors as determinants of viral host range and tropism led us to study the host cell receptor for this unusual picornavirus. We produced a monoclonal antibody (MAb), EVR1, which bound to the surface of HeLa cells and protected them against infection by EV...

Karnauchow, T. M.; Tolson, D. L.; Harrison, B. A.; Altman, E.; Lublin, D. M.; Dimock, K.

1996-01-01

177

Human DNA helicase V, a novel DNA unwinding enzyme from HeLa cells.  

OpenAIRE

Using a strand-displacement assay with 32P labeled oligonucleotide annealed to M13 ssDNA we have purified to apparent homogeneity and characterized a novel DNA unwinding enzyme from HeLa cell nuclei, human DNA helicase V (HDH V). This is present in extremely low abundance in the cells and has the highest turnover rate among other human helicases. From 300 grams of cultured cells only 0.012 mg of pure protein was isolated which was free of DNA topoisomerase, ligase, nicking and nuclease activi...

Tuteja, N.; Rahman, K.; Tuteja, R.; Falaschi, A.

1993-01-01

178

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

International Nuclear Information System (INIS)

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

179

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

180

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)

181

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

182

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

International Nuclear Information System (INIS)

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

183

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

International Nuclear Information System (INIS)

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

184

Rose Bengal acetate photodynamic therapy (RBAc-PDT) induces exposure and release of Damage-Associated Molecular Patterns (DAMPs) in human HeLa cells.  

Science.gov (United States)

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

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

2014-01-01

185

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

186

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

International Nuclear Information System (INIS)

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

187

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

188

Inhibition of HeLa cell DNA topoisomerase I by ATP and phosphate.  

OpenAIRE

The relaxation activity of DNA topoisomerase I from HeLa cell nuclei is strongly inhibited by a variety of purine nucleotides in the presence but not absence of 1 mM potassium phosphate. For ATP, 3-4 mM causes nearly complete inhibition. The 2'-and 3'-AMP isomer are active as well in the presence of 1 mM phosphate, but the 5'-AMP isomer and adenosine are inert. At 3 mM ATP, the titration curve for phosphate is sigmoidal with inhibition beginning abruptly at about 0.5 mM. The negatively-superc...

Low, R. L.; Holden, J. A.

1985-01-01

189

Radiobiological studies of human hybrid cells (skin fibroblasts x HeLa) and their tumourigenic segregants  

International Nuclear Information System (INIS)

Data are presented on the survival characteristics following ?-irradiation of a human hybrid cell line (skin fibroblasts x HeLa), and its tumourigenic segregant, which indicate that transition from the non-tumourigenic to the tumourigenic state is associated with a decrease in the ability to accumulate and repair sublethal damage. Previous studies have demonstrated that this transition to the tumourigenic state is also associated with loss of specific chromosomes (one copy each of chromosomes 11 and 14). It is suggested that this loss of chromosomes may account for the change in radiosensitivity. (author)

190

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

Science.gov (United States)

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

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

2010-01-25

191

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

Directory of Open Access Journals (Sweden)

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

192

Spontaneous premature chromosome condensation, micronucleus formation, and non-apoptotic cell death in heated HeLa S3 cells. Ultrastructural observations.  

OpenAIRE

Hyperthermia is an efficient means of inducing cell death in vivo and in vitro. Among human neoplastic cells, HeLa S3 cells are susceptible to heat injury when exposed to long duration moderate hyperthermia (41.5 C), conditions that are reproducible and sustainable in the clinical setting. Hence, HeLa S3 cells are a useful substrate for evaluation of hyperthermic injury in human neoplasia. Previous studies have demonstrated a consistent response of HeLa S3 cells to moderate hyperthermia: spon...

Swanson, P. E.; Carroll, S. B.; Zhang, X. F.; Mackey, M. A.

1995-01-01

193

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 some c...

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

1989-01-01

194

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

OpenAIRE

Serum-free cell culture methods are now routinely support mammalian cell growth, a practice adopted for ethical, scientific and safety concerns. The HeLa-S3 cell line is a subclone of the HeLa cell line that can grow in Serum-Free Medium (SFM) as well as suspension cultures. In order to optimize its culturing conditions in SFM, the present study investigated the efficacy of insulin and L-glutamine additives as biotic factors as well as osmotic stress as abiotic factor, all affecting growth ki...

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

2011-01-01

195

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

OpenAIRE

HeLa 229 cells were treated with methotrexate (MTX) and doxorubicin (DOX), utilizing fourth generation (G4), amine terminated poly(amidoamine) {PAMAM} dendrimer as the drug carrier. In vitro kinetic studies of the release of both MTX and DOX in presence and absence of G4, amine terminated PAMAM dendrimers suggest that controlled drug release can be achieved in presence of the dendrimers. The cytotoxicity studies indicated improved cell death by dendrimer-drug combination, compared to the cont...

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

2009-01-01

196

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

Directory of Open Access Journals (Sweden)

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

197

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

International Nuclear Information System (INIS)

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

198

Activation of P2Y2 purinoceptor inhibits the activity of the Na+/K+-ATPase in HeLa cells.  

Science.gov (United States)

The role of ATP on regulation of the Na(+)/K(+)-ATPase activity in the human cancerous HeLa cells was investigated. HeLa cells stimulated with increasing ATP concentrations showed a dose-dependent inhibition of the Na(+)/K(+)-ATPase activity. These effects were also obtained by UTP. ATP and UTP provoked a rise in intracellular calcium concentration ([Ca(2+)](i)) persisting for at least 4 min. The inhibitor of phospholipase C, U73122, blocked the elevation of [Ca(2+)](i) provoked by ATP/UTP. The expression of mRNA for P2Y2 and P2Y6 receptors was demonstrated by RT-PCR. ATP/UTP activated PKC-alpha, -betaI and -epsilon isoforms, but not PKC-delta and -zeta. The inhibition of the Na(+)/K(+)-ATPase activity by ATP/UTP was blocked by Gö6976, a specific inhibitor of the calcium-dependent PKCs. In conclusion, our results suggest that ATP/UTP modulate Na(+)/K(+)-ATPase activity in HeLa cells through the P2Y2 purinoceptor via calcium mobilisation and activation of calcium-dependent PKCs. PMID:12401526

Muscella, Antonella; Elia, Maria Giovanna; Greco, Simona; Storelli, Carlo; Marsigliante, Santo

2003-01-01

199

Branched dimerization of Tat peptide improves permeability to HeLa and hippocampal neuronal cells.  

Science.gov (United States)

A dimeric branched peptide TATp-D designed as an analogue of the HIV-Tat protein transduction domain (TATp), a prototypical cell penetrating peptide (CPP), demonstrates significantly enhanced cell uptake at 0.25 to 2.5 ?M. Live cell confocal laser scanning microscopy revealed that multivalency dramatically improved the permeation potency of TATp-D to HeLa and primary hippocampal neuronal cells. The observed enhanced ability of TATp-D to translocate through the membrane is highlighted by a non-linear dependence on concentration, exhibiting the greatest uptake at sub-micromolar concentrations as compared to TATp. Multimerization via bis-Fmoc Lysine offered a synthetically straightforward method to investigate the effects of multivalent CPPs while offering orthogonal handles for cargo attachment, increasing the utility of CPPs at significantly lower concentrations. PMID:25733181

Monreal, I Abrrey; Liu, Qian; Tyson, Katherine; Bland, Tyler; Dalisay, Doralyn S; Adams, Erin V; Wayman, Gary A; Aguilar, Hector C; Saludes, Jonel P

2015-03-12

200

Hyperthermia enhances the reactivation of irradiated adenovirus in HeLa cells  

International Nuclear Information System (INIS)

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

201

The Ubiquitin Ligase UBE3A Dampens ERK Pathway Signalling in HPV E6 Transformed HeLa Cells  

Science.gov (United States)

Signalling through the ERK MAP kinase pathway plays an important role in many biological processes and it is often deregulated in disease states such as cancer. One major effect of MAP kinase signalling is to promote gene expression through the phosphorylation and activation of transcription factors like ELK1. ELK1 in turn controls the activity of immediate-early genes such as FOS. Here we have used ELK1 activation in HeLa cells as a read out to conduct a genome-wide siRNA screen to identify negative regulators of ERK-mediated immediate-early gene activation. One of the candidates that we identified was the E3 ubiquitin ligase UBE3A/E6-AP. Reductions in UBE3A levels cause increased basal levels of ERK activity, a loss of growth factor-mediated ERK activation and concomitant defects in immediate-early gene expression. Thus, UBE3A acts to dampen down basal level ERK activation and to prime the pathway for growth factor-mediated activation. Mechanistically, we demonstrate that UBE3A functions in HeLa cells through its binding partner, HPV18 E6 protein and the E6 target protein p53. Loss of either E6 or p53 blocks the effect of UBE3A depletion on ERK pathway signalling, indicating that in the context of oncogenic viral protein expression, UBE3A plays an important role in negating the consequences of p53 activation on ERK pathway signalling. PMID:25815718

Aguilar-Martinez, Elisa; Morrisroe, Claire; Sharrocks, Andrew D.

2015-01-01

202

Evidence for a role of MRCK in mediating HeLa cell elongation induced by the C1 domain ligand HMI-1a3.  

Science.gov (United States)

Diacylglycerol (DAG) is a central mediator of signaling pathways that regulate cell proliferation, survival and apoptosis. Therefore, C1 domain, the DAG binding site within protein kinase C (PKC) and other DAG effector proteins, is considered a potential cancer drug target. Derivatives of 5-(hydroxymethyl)isophthalic acid are a novel group of C1 domain ligands with antiproliferative and differentiation-inducing effects. Our previous work showed that these isophthalate derivatives exhibit antiproliferative and elongation-inducing effects in HeLa human cervical cancer cells. In this study we further characterized the effects of bis(3-trifluoromethylbenzyl) 5-(hydroxymethyl)isophthalate (HMI-1a3) on HeLa cell proliferation and morphology. HMI-1a3-induced cell elongation was accompanied with loss of focal adhesions and actin stress fibers, and exposure to HMI-1a3 induced a prominent relocation of cofilin-1 into the nucleus regardless of cell phenotype. The antiproliferative and morphological responses to HMI-1a3 were not modified by pharmacological inhibition or activation of PKC, or by RNAi knock-down of specific PKC isoforms, suggesting that the effects of HMI-1a3 were not mediated by PKC. Genome-wide gene expression microarray and gene set enrichment analysis suggested that, among others, HMI-1a3 induces changes in small GTPase-mediated signaling pathways. Our experiments revealed that the isophthalates bind also to the C1 domains of ?2-chimaerin, protein kinase D (PKD) and myotonic dystrophy kinase-related Cdc42-binding kinase (MRCK), which are potential mediators of small GTPase signaling and cytoskeletal reorganization. Pharmacological inhibition of MRCK, but not that of PKD attenuated HMI-1a3-induced cell elongation, suggesting that MRCK participates in mediating the effects of HMI-1a3 on HeLa cell morphology. PMID:24486483

Talman, Virpi; Gateva, Gergana; Ahti, Marja; Ekokoski, Elina; Lappalainen, Pekka; Tuominen, Raimo K

2014-05-13

203

Growth of human rhinovirus in H1-HeLa cell suspension culture and purification of virions.  

Science.gov (United States)

HeLa cell culture is the most widely used system for in vitro studies of the basic biology of human rhinovirus (HRV). It is also useful for making sufficient quantities of virus for experiments that require highly concentrated and purified virus. This chapter describes the protocols for producing a large amount of HeLa cells in suspension culture, using these cells to grow a large quantity of virus of HeLa-adapted HRV-A and -B serotypes, and making highly concentrated virus stock and highly purified virions. These purified HRV virions are free of cellular components and suitable for experiments that are sensitive to cellular contaminations. PMID:25261306

Lee, Wai-Ming; Chen, Yin; Wang, Wensheng; Mosser, Anne

2015-01-01

204

Tat protein of human immunodeficiency virus type 1 represses expression of manganese superoxide dismutase in HeLa cells.  

OpenAIRE

Using a HeLa cell line stably transfected with the tat gene from human immunodeficiency virus type 1, we have found that the expression of the regulatory Tat protein suppresses the expression of cellular Mn-containing superoxide dismutase (Mn-SOD). This enzyme is one of the cell's primary defenses against oxygen-derived free radicals and is vital for maintaining a healthy balance between oxidants and antioxidants. The parental HeLa cells expressed nearly equivalent amounts of Cu,Zn- and Mn-SO...

Flores, S. C.; Marecki, J. C.; Harper, K. P.; Bose, S. K.; Nelson, S. K.; Mccord, J. M.

1993-01-01

205

PVA engineered microcapsules for targeted delivery of camptothecin to HeLa cells  

International Nuclear Information System (INIS)

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

206

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. PMID:24038446

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

2015-02-01

207

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)

208

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

International Nuclear Information System (INIS)

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

209

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

International Nuclear Information System (INIS)

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

210

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

OpenAIRE

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

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

2009-01-01

211

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

212

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

Directory of Open Access Journals (Sweden)

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

ANA NICIFOROVIC

2007-10-01

213

Abrus precatorius agglutinin-derived peptides induce ROS-dependent mitochondrial apoptosis through JNK and Akt/P38/P53 pathways in HeLa cells.  

Science.gov (United States)

10kDAGP, a tryptic digest of Abrus precatorius lectin 'Agglutinin' is known to induce apoptosis by mitochondria-dependent pathways in human cervical cancer (HeLa) cells. The present study was focused on deciphering the detailed molecular mechanism of apoptosis induction in vitro by 10kDAGP and also its in vivo therapeutic efficacy. For in vivo model, HeLa cell encapsulated hollow fiber was implanted in Swiss Albino mice and treated with 10kDAGP. Our results showed that 10kDAGP was able to enter the cell within a span of 20min and co-localized with mitochondria after 90min. of incubation. A drastic loss of mitochondrial membrane potential was noted within 6h of 10kDAGP administration along with an increase in ROS generation. ROS further led to symptoms of early apoptosis by deregulating Akt (Protein Kinase B) and activating c-Jun N-terminal Kinase (JNK), p38 Mitogen Activated Protein Kinase (MAPK), p53, and autophagy starting from ?8h of incubation. Besides in vitro conditions, 10kDAGP activated JNK to mediate cancer cell killing in vivo. Therefore, 10kDAGP can be an excellent therapeutic agent as it can act through different ways in the cellular system. Future studies are directed to screen out active peptides from the pool of peptides and to study whether the mode of action is in synergistic way or in individual forms. PMID:25305377

Behera, Birendra; Mishra, Debasish; Roy, Bibhas; Devi, K Sanjana P; Narayan, Rajan; Das, Joyjyoti; Ghosh, Sudip K; Maiti, Tapas K

2014-10-01

214

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

International Nuclear Information System (INIS)

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

215

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

International Nuclear Information System (INIS)

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

216

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

International Nuclear Information System (INIS)

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

217

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

International Nuclear Information System (INIS)

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

218

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

Science.gov (United States)

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

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

2014-01-01

219

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

220

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

221

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

Science.gov (United States)

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

Kishimoto, Takuji; Fukuzawa, Yoichiro; Tada, Manabu

1990-09-01

222

Melatonin protects against rotenone-induced cell injury via inhibition of Omi and Bax-mediated autophagy in Hela cells.  

Science.gov (United States)

Parkinson's disease is the second most common neurodegenerative disease, and environmental toxins such as rotenone play an important role in causing degeneration of dopaminergic neurons. Melatonin, a major secretory product of pineal, is recently reported to protect against rotenone-induced cell death in animal models. Yet, the mechanism involved in this protection needs to be elucidated. Here, we report that rotenone treatment (0-100 ?M) decreased cell survival of Hela cells in a dose-dependent manner. At concentrations ranging from 0.1 to 100 ?M, rotenone induced a dose-dependent increase in the expression of microtubule-associated protein 1 light chain 3 (LC3)-II, a protein associated with the autophagosomal membrane. Knockdown of Bax or Omi using shRNA inhibited 1 ?M rotenone-induced autophagy. To determine whether melatonin would protect cells against rotenone-induced cell death and autophagy, we pretreated Hela cells with 250 ?M melatonin for 24 hr in the presence of rotenone. Melatonin inhibited Bax expression and the release of the omi/HtrA2 into the cytoplasm induced by 1 ?M rotenone. Melatonin 250 ?M treatment also suppressed cell death induced by 0.1-100 ?M rotenone and protected against the formation of LC3-II in cells exposed to 1 ?M rotenone. This work demonstrates a novel role for melatonin as a neuroprotective agent against rotenone. PMID:21883444

Zhou, Hongyan; Chen, Jie; Lu, Xilin; Shen, Cunzhou; Zeng, Jinsheng; Chen, Ling; Pei, Zhong

2012-01-01

223

Lethal Effects of Radiation and Platinum Analogues on Multicellular Spheroids of HeLa Cells  

International Nuclear Information System (INIS)

Multicellul ar tumor spheroids of HeLa cells have been grown in a static culture system. Samples of spheroids were exposed for 2 h to graded concentration of sis-platinum and its analogue, carboplatin, and then response assayed by survival of clonogenic cells. The purpose of present experiment is to clarify the effectiveness of these platinum compounds and to evaluate intrinsic radiosensitivity of cells using spheroids of HeLa cells as an experimental in vitro model. Variations of the drug sensitivity of monolayers as well as spheroids were also evaluated in cell-survival curves. In cia-platinum concentration-survival cutie, there was a large shoulder extending as far as Cq=3.4 mM, after which there was exponential decrease in survival curve having a Co Value of 1,2 ?M in spheroids. While the Co for the spheroids was essentially no significant change, but Cq value was larger than that of monolayers. This suggest that the effect of cis-platinum is greater in the monolayer with actively proliferating cells than hypoxic one. In the carboplatin concentration-survival curves, the Co value of spheroids was 15.0 mM and the ratio with the Co from monolayer cell (32.5 mM) was 0.46, thus indicating that the spheroids had a greater sensitivity to carboplatin than monolayers. Therefore, the effect of carboplatin is mainly on the deeper layers of spheroids acting as hypoxic cell sensitizer. The enhanced effect was obtained for monolayer cells using combined X-ray and carboplatin lls using combined X-ray and carboplatin treatment 2 hours before irradiation. The result shown in isobologram analysis for the level of surviving fraction at 0.01 indicated that the effect of two agents was truly supra-additive. From this experimental data, carboplatin has excited much receipt interest as one of the most promising, since it is almost without nephrotoxicity and causes less gastrointestinal toxicity than cia-platinum. Interaction between carboplatin and radiation might play an important role for more effective local tumor control

224

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

225

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

Science.gov (United States)

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

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

2009-06-01

226

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

Directory of Open Access Journals (Sweden)

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

Bazowski Jessa

2010-10-01

227

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

Directory of Open Access Journals (Sweden)

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

Abdalla A. El Shereef

2011-01-01

228

Segregation of nucleolar components coincides with caspase-3 activation in cisplatin-treated HeLa cells.  

Czech Academy of Sciences Publication Activity Database

Ro?. 114, ?. 4 (2001), s. 663-670. ISSN 0021-9533 R&D Projects: GA ?R GV521/96/K117 Institutional research plan: CEZ:AV0Z5038910 Keywords : nucleolar components * caspase-3 activation * treated HeLa cells Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 6.213, year: 2001

Horký, M.; Wurzer, G.; Kotala, Vladimír; Anton, M.; Vojt?šek, B.; Vácha, J.; Wesierska-Gadek, J.

2001-01-01

229

DNA synthesis in generation 1 in x-irradiated HeLa cells  

International Nuclear Information System (INIS)

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

230

Acid polypeptides as inhibitors of the repair of double-strand DNA breaks induced by ?-irradiation of Hela cells  

International Nuclear Information System (INIS)

The effect of natural modificator's synthetic analogue -polyaspartylglytamate (AG) - on the repair of radiation-induced double-strand DNA breaks is studies. The radiation and modificator effects were determined by the criterion of the formation of chromosome recombinations and reproductive death of cells on Hela cell culture and in Chinese hamsters. It is shown that the incubation of Hela cells with AG doubles and triples the degradation effect of rdiation at 50 and 10 Gy doses. When radiation dose equals 1 Gy and repair time is G-22 h, 1.5 - 3 time - increased yield of chromotide and chromosome abberations is detected in Chinese hamster cells in the presence of the modificator during all periods of cell fixation. The effect of radiation mutagenic action enhancement by the modificator is not observed during the incubation of cells with AG 30-45 min after irradiation

231

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

Energy Technology Data Exchange (ETDEWEB)

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

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

2008-11-10

232

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

International Nuclear Information System (INIS)

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

233

Nitric oxide enhances extracellular ATP induced Ca²? oscillation in HeLa cells.  

Science.gov (United States)

Calcium (Ca(2+)) oscillations play a central role in varieties of cellular processes including fertilization and immune response, but controversy over the regulation mechanisms still exists. It has been known that nitric oxide (NO) dependently regulates Ca(2+) signaling in most physiopathological processes. Previous study indicated that eNOS translocation during some pathological process influences intracellular Ca(2+) homeostasis. In this study, we investigated the role and mechanism of NO on Ca(2+) release by overexpressing eNOS in cytoplasm (Cyto-eNOS) and endoplasmic reticulum (ER-eNOS) of HeLa cells. We found that the properties of Ca(2+) release were altered by the overexpression of eNOS. The amplitude and frequency of extracellular ATP (eATP)-induced Ca(2+) oscillation were enhanced in both Cyto-eNOS and ER-eNOS cells, respectively. Especially, the enhancement of the amplitude and frequency of the Ca(2+) oscillation was much more significant in the ER-eNOS cells than that of Cyto-eNOS cells. Further study indicated that this effect was abrogated by NO inhibitor, L-NAME, suggesting it was not an artificial result induced by ER stress. Furthermore, an up-regulated phosphorylation of phospholamban (PLB) was observed and the sarco-endoplasmic reticulum Ca(2+)-ATPase (SERCA) function was activated followed by the significant increase in the ER Ca(2+) load. Taken together, we revealed a novel regulatory mechanism of Ca(2+) oscillation. PMID:25461674

Tang, Yinglong; Yin, Yin; Miao, Lin; Wei, Bin; Zhai, Kui; Ji, Guangju

2015-01-01

234

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

235

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)

236

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

Science.gov (United States)

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

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

2014-09-01

237

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

Science.gov (United States)

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

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

2013-11-01

238

A novel dithiocarbamate derivative induces cell apoptosis through p53-dependent intrinsic pathway and suppresses the expression of the E6 oncogene of human papillomavirus 18 in HeLa cells.  

Science.gov (United States)

Dithiocarbamates (DTCs) exhibit a broad spectrum of antitumor activities, however, their molecular mechanisms of antitumor have not yet been elucidated. Previously, we have synthesized a series of novel dithiocarbamate derivatives. These DTCs were examined for cytotoxic activities against five human cancer cell lines. In this study, one of dithiocarbamate (DTC1) with higher potential for HeLa cells was chosen to investigate molecular mechanisms for its anti-tumor activities. DTC1 could inhibit proliferation, and highly induce apoptosis in HeLa cells by activating caspase-3, -6 and -9; moreover, activities of caspase-3, -6 and -9 were inhibited by pan-caspase inhibitor, Z-VAD-FMK. Furthermore, DTC1 decreased the levels of Bcl-2 and Bcl-xL, and increased expression of cytosol cytochrome c, Bak, Bax and p53 in a time-dependent manner but had no effect on the level of Rb. It was shown that DTC1 induced HeLa cells apoptosis through a p53-dependent pathway as tested by the wild type p53 inhibitor, pifithrin-?. Additionally, the relative expression of E6 and E7 were evaluated in HPV18-positive (HeLa cells) by real-time PCR and western blotting. The results firstly demonstrated that DTC1 suppressed both expression of E6 mRNA and E6 oncoprotein, but had no effect on the expression of E7 mRNA and protein in HPV18. Our results suggested that DTC1 may serve as novel chemotherapeutic agents in the treatment of cervical cancer and potential anti-HPV virus candidates that merit further studies. PMID:25772545

Li, Yanhong; Qi, Hongxue; Li, Xiaobo; Hou, Xueling; Lu, Xueying; Xiao, Xiangwen

2015-06-01

239

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  

Scientific Electronic Library Online (English)

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.

240

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)

241

Bioengineering functional copolymers. XII. Interaction of boron-containing and PEO branched derivatives of poly(MA-alt-MVE with HeLa cells  

Directory of Open Access Journals (Sweden)

Full Text Available Novel boron-containing bioengineering copoly- mer and its ?-hydoxy-?-methoxy-poly(ethy- lene oxide (PEO macrobranched derivatives were synthesized by (1 partially amidolysis of poly(maleic anhydride-alt-methyl vinyl ether w?th ethanolamine ester of diphenylboronic acid and (2 esterification of synthesized B-con- taining copolymers with PEO. They had a com- bination of hydrophilic/hydrophobic linkages, free carboxylic groups, positive charges and an ionized organoboron linkage as antitumor sites, along with an ability to interact with HeLa cells. The structure, composition and properties (cy- totoxicity and antitumor activity of synthe- sized copolymers were investigated. In vitro cytotoxicity results, obtained by the fluore scence microscopy measurements indicate that unlike the virgin copolymer, boron-containing and PEO macrobranched derivatives exhibit higher antitumor activity. It was found that organoboron copolymer exhibits the most apo- ptotic and necrotic effects against HeLa cells whereas a minor effect relative to cancer cells was observed on L929 Fibroblast cells.

Gülcihan Kurucu

2010-01-01

242

Gallic acid reduces cell viability, proliferation, invasion and angiogenesis in human cervical cancer cells  

OpenAIRE

Gallic acid is a trihydroxybenzoic acid, also known as 3,4,5-trihydroxybenzoic acid, which is present in plants worldwide, including Chinese medicinal herbs. Gallic acid has been shown to have cytotoxic effects in certain cancer cells, without damaging normal cells. The objective of the present study was to determine whether gallic acid is able to inhibit human cervical cancer cell viability, proliferation and invasion and suppress cervical cancer cell-mediated angiogenesis. Treatment of HeLa...

Zhao, Bing; Hu, Mengcai

2013-01-01

243

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

International Nuclear Information System (INIS)

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

244

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

International Nuclear Information System (INIS)

[3H]Ouabain specifically bound at sublethal concentrations to Na,K-ATPase on the surface of HeLa cells is taken up (internalized) by the cells at a rate of three membrane equivalents of labeled sites per generation. Immediately following a pulse label with the glycoside, codistribution of radioactivity with the surface marker 5'-nucleotidase is found in both conventional sucrose-gradient fractionation and in fractionation following a digitonin treatment. At appropriate concentrations digitonin increases the buoyant density of the HeLa surface membrane and solubilizes the lysosomal marker ?-hexosaminidase (Tulkens et al., 1974). After internalization, [3H]ouabain is also solubilized by digitonin. A shear analysis is described which shows internalized ouabain and ?-hexosaminidase to be codistributed in a particulate fraction that is homogeneous with respect to shear; extrapolation to zero-shear shows that little or none of either marker is found in the soluble fraction of the cytosol. Both markers are coreleased from the particulate fraction by osmotic shock. Although internalized ouabain is subsequently released from these cells with a half-time of about 70 hr, apparently by exocytosis, the shear sensitivity of the remaining cell-associated ouabain does not change for up to 72 hr. Thus ouabain (together with Na,K-ATPase.) appears to be taken up from the surface into a lysosomal compartment and, by at least one criterion, this compartment does no one criterion, this compartment does not change its physical properties with time, i.e., does not ''age.''

245

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

International Nuclear Information System (INIS)

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

246

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

Scientific Electronic Library Online (English)

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

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

2003-12-01

247

Opposite replication polarity of the germ line c-myc gene in HeLa cells compared with that of two Burkitt lymphoma cell lines.  

OpenAIRE

To study the cell type specificity of the direction of replication of the human c-myc genes and the relationship of replication polarity to transcriptional activity, we analyzed the directions of replication of the c-myc genes in two Burkitt lymphoma cell lines, CA46 and ST486, and in HeLa cells. On the basis of in vitro runoff replication of forks initiated in intact cells, we found that transcribed c-myc genes in the germ line configuration in HeLa cells were replicated in the direction of ...

Leffak, M.; James, C. D.

1989-01-01

248

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

OpenAIRE

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

249

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

OpenAIRE

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 molec...

1987-01-01

250

Triton X-100 concentration effects on membrane permeability of a single HeLa cell by scanning electrochemical microscopy (SECM)  

OpenAIRE

Changes in HeLa cell morphology, membrane permeability, and viability caused by the presence of Triton X-100 (TX100), a nonionic surfactant, were studied by scanning electrochemical microscopy (SECM). No change in membrane permeability was found at concentrations of 0.15 mM or lower during an experimental period of 30 to 60 min. Permeability of the cell membrane to the otherwise impermeable, highly charged hydrophilic molecule ferrocyanide was seen starting at concentrations of TX100 of abo...

Koley, Dipankar; Bard, Allen J.

2010-01-01

251

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

Science.gov (United States)

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 synthesized q-base to q-deficient cells. While the q-base was always inserted into tRNA under aerobic conditions, HeLa cells lost this ability under hypoxic conditions, however, only when serum factors became depleted from the culture medium. The inability to insert q into tRNA did not result from a lack of substrate, because the q-base accumulated within these cells against the concentration gradient, suggesting the presence of an active transport system for this base in HeLa cells. The activity of the TGT enzyme was restored after treatment of the cells with the protein kinase C activator, TPA, even in the presence of mRNA or protein synthesis inhibitors. The results indicate that the eukaryotic tRNA modifying enzyme, TGT, is a downstream target of activated protein kinase C. PMID:7630726

Langgut, W; Reisser, T

1995-07-11

252

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 ...

253

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

Science.gov (United States)

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 had a spatial resolution at or slightly below a micron (900nm), as defined by the distance between the 16 and 84% intensities in a line scan across the edge of the features. Additionally, important parameters influencing the quality of the HeLa cell 3D images were investigated. Atomic force microscopy measurements revealed that the HeLa cells were sputtered at a rate of approximately 4 nm per 1013 C60+ ions/cm2 at 10 keV and a 45° incident angle. Optimal 3D images were acquired using a Bi3+ liquid metal ion gun operating in the simultaneous high mass and spatial resolution mode. PMID:24131300

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

2013-01-01

254

Combination of cytokinin and auxin induces apoptosis, cell cycle progression arrest and blockage of the Akt pathway in HeLa cells.  

Science.gov (United States)

Plant cytokinins and auxins have recently been proposed as novel cancer therapies, which proceed via different mechanisms; however, their combined use has not been investigated. To the best of our knowledge, the present study was the first to show that the cytokinin ortho?methoxytopolin?riboside (MeoTR) strongly inhibited the proliferation of HeLa cells, the effect of which was synergistically enhanced by auxin indole?3?acetic acid (IAA), while IAA demonstrated to have no cytotoxic effects on cells. MeoTR was found to activate intrinsic and extrinsic caspase?dependent pathways, and IAA potentiated this activation. In addition, these effects were blocked by Z?Val?Ala?Asp?fluoromethylketone (Z?VAD?FMK), a pan?specific?caspase?inhibitor. IAA increased the MeoTR? induced inhibition of B cell lymphoma 2 (Bcl?2) and survivin, whereas IAA?only decreased Bcl?2 expression. MeoTR downregulated phosphorylated (p)?pyruvate dehydrogenase kinase 1, p?Akt and p?glycogen synthase kinase 3?, the effect of which was more potent in combination with IAA, despite the weak effect of IAA alone. LY294002, an Akt?inhibitor, was able to increase the inhibition of p?Akt through MeoTR and combination treatment. IAA and MeoTR increased the proportion of cells in S phase independently. However, the combination treatment induced a further increase. In addition, IAA and MeoTR treatment downregulated protein levels of cyclin A, cyclin?dependent kinase 2 (CDK2) and p?CDK2, and upregulated protein levels of p21 and p27. Furthermore, the combination treatment enhanced these effects, indicating that IAA potentiated the inhibitory effect of MeoTR on HeLa cells via cell cycle progression arrest and accumulation in S phase, coupled with the negative regulation of Bcl?2. In conclusion, the results of the present study suggested that treatment with these two phytohormones in combination, may offer a novel therapeutic strategy for the treatment of malignant cervical cancer. PMID:25738331

Zhao, Liwei; Liu, Peng; Guo, Guangqin; Wang, Li

2015-07-01

255

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

International Nuclear Information System (INIS)

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

256

Selective eradication of cancer cells in vitro  

International Nuclear Information System (INIS)

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

257

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

Directory of Open Access Journals (Sweden)

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

Halder Rashi

2012-03-01

258

Organellar proteome analyses of ricin toxin-treated HeLa cells.  

Science.gov (United States)

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

259

Identification of proteins associated with Aha1 in HeLa cells by quantitative proteomics.  

Science.gov (United States)

The identification of the activator of heat shock protein 90 (Hsp90) ATPase's (Aha1) protein-protein interaction (PPI) network will provide critical insights into the relationship of Aha1 with multi-molecular complexes and shed light onto Aha1's interconnections with Hsp90-regulated biological functions. Flag-tagged Aha1 was over-expressed in HeLa cells and isolated by anti-Flag affinity pull downs, followed by trypsin digestion and identification co-adsorbing proteins by liquid chromatography-tandem mass spectroscopy (LC-MS/MS). A probability-based identification of Aha1 PPIs was generated from the LC-MS/MS analysis by using a relative quantification strategy, spectral counting (SC). By comparing the SC-based protein levels between Aha1 pull-down samples and negative controls, 164 Aha1-interacting proteins were identified that were quantitatively enriched in the pull-down samples over the controls. The identified Aha1-interacting proteins are involved in a wide number of intracellular bioprocesses, including DNA maintenance, chromatin structure, RNA processing, translation, nucleocytoplasmic and vesicle transport, among others. The interactions of 33 of the identified proteins with Aha1 were further confirmed by Western blotting, demonstrating the reliability of our affinity-purification-coupled quantitative SC-MS strategy. Our proteomic data suggests that Aha1 may participate in diverse biological pathways to facilitate Hsp90 chaperone functions in response to stress. PMID:25614414

Sun, Liang; Hartson, Steven D; Matts, Robert L

2015-05-01

260

Interactions of Campylobacter jejuni cytolethal distending toxin subunits CdtA and CdtC with HeLa cells.  

Science.gov (United States)

Campylobacter jejuni produces a toxin, called cytolethal distending toxin (CDT), which causes direct DNA damage leading to invocation of DNA damage checkpoint pathways. The affected cells arrest in G(1) or G(2) and eventually die. CDT consists of three protein subunits, CdtA, CdtB, and CdtC, with CdtB recently identified as a nuclease. However, little is known about the functions of CdtA or CdtC. In this work, enzyme-linked immunosorbent assay-based experiments were used to show, for the first time, that both CdtA and CdtC bound with specificity to the surface of HeLa cells, whereas CdtB did not. Varying the order of the addition of subunits for reconstitution of the holotoxin had no effect on activity. In addition, mutants containing deletions of conserved regions of CdtA and CdtC were able to bind to the surface of HeLa cells but were not able to participate in holotoxin assembly. Finally, both Cdt mutant subunits were able to effectively compete with CDT holotoxin in the HeLa cell binding assay. PMID:12933829

Lee, Robert B; Hassane, Duane C; Cottle, Daniel L; Pickett, Carol L

2003-09-01

261

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

Directory of Open Access Journals (Sweden)

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

Ajay Amrendra K

2012-01-01

262

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

Energy Technology Data Exchange (ETDEWEB)

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

263

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

International Nuclear Information System (INIS)

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

264

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

OpenAIRE

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

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

1989-01-01

265

Single Hepatitis-B Virus Core Capsid Binding to Individual Nuclear Pore Complexes in HeLa Cells  

OpenAIRE

We investigate the interaction of hepatitis B virus capsids lacking a nuclear localization signal with nuclear pore complexes (NPCs) in permeabilized HeLa cells. Confocal and wide-field optical images of the nuclear envelope show well-spaced individual NPCs. Specific interactions of capsids with single NPCs are characterized by extended residence times of capsids in the focal volume which are characterized by fluorescence correlation spectroscopy. In addition, single-capsid-tracking experimen...

Lill, Yoriko; Lill, Markus A.; Fahrenkrog, Birthe; Schwarz-herion, Kyrill; Paulillo, Sara; Aebi, Ueli; Hecht, Bert

2006-01-01

266

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

Directory of Open Access Journals (Sweden)

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

267

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

International Nuclear Information System (INIS)

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

268

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

Science.gov (United States)

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.

Atif, M.; Firdous, S.; Khurshid, A.; Noreen, L.; Zaidi, S. S. Z.; Ikram, M.

2009-12-01

269

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

270

Squamous cell skin cancer  

Science.gov (United States)

... cell; NMSC - squamous cell; Squamous cell skin cancer; Squamous cell carcinoma of the skin ... squamous cell cancer is called Bowen disease (or squamous cell carcinoma in situ). This type does not spread to ...

271

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

272

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

273

Multi-color imaging of fluorescent nanodiamonds in living HeLa cells using direct electron-beam excitation.  

Science.gov (United States)

Multi-color, high spatial resolution imaging of fluorescent nanodiamonds (FNDs) in living HeLa cells has been performed with a direct electron-beam excitation-assisted fluorescence (D-EXA) microscope. In this technique, fluorescent materials are directly excited with a focused electron beam and the resulting cathodoluminescence (CL) is detected with nanoscale resolution. Green- and red-light-emitting FNDs were employed for two-color imaging, which were observed simultaneously in the cells with high spatial resolution. This technique could be applied generally for multi-color immunostaining to reveal various cell functions. PMID:24403210

Nawa, Yasunori; Inami, Wataru; Lin, Sheng; Kawata, Yoshimasa; Terakawa, Susumu; Fang, Chia-Yi; Chang, Huan-Cheng

2014-03-17

274

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

International Nuclear Information System (INIS)

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

275

A Stable HeLa Cell Line That Inducibly Expresses Poliovirus 2Apro: Effects on Cellular and Viral Gene Expression  

OpenAIRE

A HeLa cell clone (2A7d) that inducibly expresses the gene for poliovirus protease 2A (2Apro) under the control of tetracycline has been obtained. Synthesis of 2Apro induces severe morphological changes in 2A7d cells. One day after tetracycline removal, cells round up and a few hours later die. Poliovirus 2Apro cleaves both forms of initiation factor eIF4G, causing extensive inhibition of capped-mRNA translation a few hours after protease induction. Methoxysuccinyl-Ala-Ala-Pro-Val-chloromethy...

Barco, Angel; Feduchi, Elena; Carrasco, Luis

2000-01-01

276

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

277

Analysis of gene expression profiles in HeLa cells in response to overexpression or siRNA-mediated depletion of NASP  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background NASP (Nuclear Autoantigenic Sperm Protein is a linker histone chaperone required for normal cell division. Changes in NASP expression significantly affect cell growth and development; loss of gene function results in embryonic lethality. However, the mechanism by which NASP exerts its effects in the cell cycle is not understood. To understand the pathways and networks that may involve NASP function, we evaluated gene expression in HeLa cells in which NASP was either overexpressed or depleted by siRNA. Methods Total RNA from HeLa cells overexpressing NASP or depleted of NASP by siRNA treatment was converted to cRNA with incorporation of Cy5-CTP (experimental samples, or Cy3-CTP (control samples. The labeled cRNA samples were hybridized to whole human genome microarrays (Agilent Technologies, Wilmington, Delaware, USA. Various gene expression analysis techniques were employed: Significance Analysis of Microarrays (SAM, Expression Analysis Systematic Explorer (EASE, and Ingenuity Pathways Analysis (IPA. Results From approximately 36 thousand genes present in a total human genome microarray, we identified a set of 47 up-regulated and 7 down-regulated genes as a result of NASP overexpression. Similarly we identified a set of 56 up-regulated and 71 down-regulated genes as a result of NASP siRNA treatment. Gene ontology, molecular network and canonical pathway analysis of NASP overexpression demonstrated that the most significant changes were in proteins participating in organismal injury, immune response, and cellular growth and cancer pathways (major "hubs": TNF, FOS, EGR1, NF?B, IRF7, STAT1, IL6. Depletion of NASP elicited the changed expression of proteins involved in DNA replication, repair and development, followed by reproductive system disease, and cancer and cell cycle pathways (major "hubs": E2F8, TP53, FGF, FSH, FST, hCG, NF?B, TRAF6. Conclusion This study has demonstrated that NASP belongs to a network of genes and gene functions that are critical for cell survival. We have confirmed the previously reported interactions between NASP and HSP90, HSP70, histone H1, histone H3, and TRAF6. Overexpression and depletion of NASP identified overlapping networks that included TNF as a core protein, confirming that both high and low levels of NASP are detrimental to cell cycle progression. Networks with cancer-related functions had the highest significance, however reproductive networks containing follistatin and FSH were also significantly affected, which confirmed NASP's important role in reproductive tissues. This study revealed that, despite some overlap, each response was associated with a unique gene signature and placed NASP in important cell regulatory networks.

Alekseev Oleg

2009-05-01

278

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

279

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

280

Gallic acid reduces cell viability, proliferation, invasion and angiogenesis in human cervical cancer cells  

Science.gov (United States)

Gallic acid is a trihydroxybenzoic acid, also known as 3,4,5-trihydroxybenzoic acid, which is present in plants worldwide, including Chinese medicinal herbs. Gallic acid has been shown to have cytotoxic effects in certain cancer cells, without damaging normal cells. The objective of the present study was to determine whether gallic acid is able to inhibit human cervical cancer cell viability, proliferation and invasion and suppress cervical cancer cell-mediated angiogenesis. Treatment of HeLa and HTB-35 human cancer cells with gallic acid decreased cell viability in a dose-dependent manner. BrdU proliferation and tube formation assays indicated that gallic acid significantly decreased human cervical cancer cell proliferation and tube formation in human umbilical vein endothelial cells, respectively. Additionally, gallic acid decreased HeLa and HTB-35 cell invasion in vitro. Western blot analysis demonstrated that the expression of ADAM17, EGFR, p-Akt and p-Erk was suppressed by gallic acid in the HeLa and HTB-35 cell lines. These data indicate that the suppression of ADAM17 and the downregulation of the EGFR, Akt/p-Akt and Erk/p-Erk signaling pathways may contribute to the suppression of cancer progression by Gallic acid. Gallic acid may be a valuable candidate for the treatment of cervical cancer. PMID:24843386

ZHAO, BING; HU, MENGCAI

2013-01-01

281

Squamous cell cancer (image)  

Science.gov (United States)

Squamous cell cancer involves cancerous changes to the cells of the middle portion of the epidermal skin layer. It is ... malignant tumor, and is more aggressive than basal cell cancer, but still may be relatively slow-growing. It ...

282

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

Directory of Open Access Journals (Sweden)

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

283

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

284

Modulation of mitochondrial transcription in response to mtDNA depletion and repletion in HeLa cells  

OpenAIRE

The steady-state amounts of mitochondrial transcripts and transcription proteins were analyzed during mtDNA depletion and subsequent repletion to gain insight into the regulation of human mitochondrial gene expression. As documented previously, HeLa cells depleted of mtDNA via treatment with ethidium bromide (EB) were found to contain reduced steady-state levels of the mitochondrial transcription factor h-mtTFA. When partially mtDNA-depleted cells were cultured in the absence of EB, h-mtTFA r...

Seidel-rogol, Bonnie L.; Shadel, Gerald S.

2002-01-01

285

Analyses of differential sensitivities of synchronized Hela S3-cells to radiations and chemical carcinogens during the cell cycle  

International Nuclear Information System (INIS)

Radiation-induction and rejoining of single-strand breaks (SSBs) in the DNA of synchronized HeLa S3 cells were investigated by alkaline sucrose density gradients. We could not find any significant differences in the extent of SSBs induced in cellular DNA and in the extent ot their rejoining throughout the cell cycle, including mitosis. The cyclic variation curve of the content of non-protein sulfhydryls (NPSH) during the cell cycle is similar to that of x,ray survivals except in mitosis, although there was no close correlation between the content of apparent total sulfhydryls (APSH) and X-ray survivals. Radiation-induced mutants resistant to 8-azaguanine (8AG) occurred in higher frequency in the radio-senvitive G1-S boundary phase than in the radioresistant G1, S and early G2 phases. Further, the pre-irradiation treatment with 50 mM cysteamine prevented reproductive death and induction of 8AG-resistant mutants by X-rays throughout the cell cycle. These findings seem to indicate that there is a close correlation between the extent of lethal radiation damage to the cells and their mutability, and that sulfhydryls may play an important role as a factor governing cellular radio-sensitivity

286

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.

287

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

288

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

OpenAIRE

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 sta...

Yujiong Wang; Xiaoming Liu; Xiujing Hao; Yong Li; Xuezhang Zhou; Yuyan Zhang

2012-01-01

289

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

290

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

Directory of Open Access Journals (Sweden)

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

291

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

292

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

Science.gov (United States)

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

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

293

Roles for Endocytosis and Low pH in Herpes Simplex Virus Entry into HeLa and Chinese Hamster Ovary Cells  

OpenAIRE

Herpes simplex virus (HSV) infection of many cultured cells, e.g., Vero cells, can be initiated by receptor binding and pH-neutral fusion with the cell surface. Here we report that a major pathway for HSV entry into the HeLa and CHO-K1 cell lines is dependent on endocytosis and exposure to a low pH. Enveloped virions were readily detected in HeLa or receptor-expressing CHO cell vesicles by electron microscopy at

Nicola, Anthony V.; Mcevoy, Anna M.; Straus, Stephen E.

2003-01-01

294

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

OpenAIRE

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

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

1999-01-01

295

Establishing threshold toxicity for introducing magnetic nanoparticles into HeLa and HEK 293 cells  

Science.gov (United States)

Although iron oxide nanoparticles have been suggested as candidate in diverse applications such as drug delivery agents, contrast agents of magnetic resonance imaging, cancer treatment through hyperthermia, etc., the upper limit for safe dosage beyond which the toxicity sets in has never been studied. In this work we report quantitative studies on the percentage change in the number of cell as a function of concentration of magnetic nanoparticles. The incubation is at 37 C and lasted for 24 hours. We found that there is a critical value of particle volume fraction, above which appreciable number of cell death occurs. This critical value was found to differ in two different cell lines indicating that HEK cells are more robust against the magnetic nanoparticles.

Chen, Kezheng; Luo, Weili; Kolattukuty, Pappachan

2006-03-01

296

Silencing Bcl-2 Expression in Epithelial Cancer Cells Using “Smart” Particles  

OpenAIRE

Short interfering RNA (siRNA) targeted against anti-apoptotic Bcl-2 protein proved to knockdown its expression and trigger cancer cell death. We used degradable, pH-sensitive, comb-like [P(EAA-co-BMA)-b-PNASI-g-P(HMA-co-TMAEMA)] polymer to condense anti-Bcl-2 siRNA into “smart” particles, which proved to shuttle their cargo past the endosomal membrane and into the cytoplasm of HeLa and UM-SCC-17B cancer cells. HeLa and UM-SCC-17B cancer cells were treated with anti-Bcl-2 particles followe...

Yen-Ling Lin; Guohua Jiang; Zhaocheng Zhang; No?r, Jacques E.; Elsayed, Mohamed E. H.

2014-01-01

297

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

298

Identification of human immunodeficiency virus envelope gene sequences influencing viral entry into CD4-positive HeLa cells, T-leukemia cells, and macrophages.  

OpenAIRE

Infectious recombinant viruses were constructed from three molecularly cloned human immunodeficiency virus (HIV) strains varying in cell tropism. All recombinants showed a high infectivity titer on phytohemagglutinin-stimulated normal T lymphocytes. However, a 120-bp region of the envelope gene including the area of the V3 hypervariable loop was found to influence infectivity titer on both clone 1022 CD4-positive HeLa cells and CD4-positive CEM leukemia cells. Infectivity for macrophages was ...

Chesebro, B.; Nishio, J.; Perryman, S.; Cann, A.; O Brien, W.; Chen, I. S.; Wehrly, K.

1991-01-01

299

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

300

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

301

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

Science.gov (United States)

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

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

2012-06-01

302

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

Science.gov (United States)

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. PMID:22619531

Nazli, Caner; Ergenc, Tugba Ipek; Yar, Yasemin; Acar, Havva Yagci; Kizilel, Seda

2012-01-01

303

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

Science.gov (United States)

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

Kim, Young Mee; Cho, Moonjae

2014-04-01

304

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 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

305

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

International Nuclear Information System (INIS)

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

306

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

Science.gov (United States)

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

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

2014-01-01

307

NADH oxidase activity (NOX) and enlargement of HeLa cells oscillate with two different temperature-compensated period lengths of 22 and 24 minutes corresponding to different NOX forms  

Science.gov (United States)

NOX proteins are cell surface-associated and growth-related hydroquinone (NADH) oxidases with protein disulfide-thiol interchange activity. A defining characteristic of NOX proteins is that the two enzymatic activities alternate to generate a regular period length of about 24 min. HeLa cells exhibit at least two forms of NOX. One is tumor-associated (tNOX) and is inhibited by putative quinone site inhibitors (e.g., capsaicin or the antitumor sulfonylurea, LY181984). Another is constitutive (CNOX) and refractory to inhibition. The periodic alternation of activities and drug sensitivity of the NADH oxidase activity observed with intact HeLa cells was retained in isolated plasma membranes and with the solubilized and partially purified enzyme. At least two activities were present. One had a period length of 24 min and the other had a period length of 22 min. The lengths of both the 22 and the 24 min periods were temperature compensated (approximately the same when measured at 17, 27 or 37 degrees C) whereas the rate of NADH oxidation approximately doubled with each 10 degrees C rise in temperature. The rate of increase in cell area of HeLa cells when measured by video-enhanced light microscopy also exhibited a complex period of oscillations reflective of both 22 and 24 min period lengths. The findings demonstrate the presence of a novel oscillating NOX activity at the surface of cancer cells with a period length of 22 min in addition to the constitutive NOX of non-cancer cells and tissues with a period length of 24 min.

Wang, S.; Pogue, R.; Morre, D. M.; Morre, D. J.

2001-01-01

308

Dual effects of 2-deoxyglucose on synthesis of the glycoprotein hormone common alpha-subunit in butyrate-treated HeLa cells.  

OpenAIRE

Sodium butyrate (Btr) (3 mM) causes a 10-fold increase in production of the glycoprotein hormone alpha-subunit in HeLa cells. The following report demonstrates that this response could be inhibited about 95% by 5 mM 2-deoxy-D-glucose (dGlc), whereas alpha-subunit production in uninduced cells was affected little or not at all. Addition of D-mannose restored the Btr induction of Hela-alpha in cultures that had been treated with dGlc. When the alpha-subunits secreted by cells cultured in Btr pl...

Cox, G. S.; Mcclure, D. S.; Cosgrove, D. E.

1987-01-01

309

Cleavage of Eukaryotic Translation Initiation Factor 4G by Exogenously Added Hybrid Proteins Containing Poliovirus 2Apro in HeLa Cells: Effects on Gene Expression  

OpenAIRE

Efficient cleavage of both forms of eukaryotic initiation factor 4G (eIF4G-1 and eIF4G-2) has been achieved in HeLa cells by incubation with hybrid proteins containing poliovirus 2Apro. Entry of these proteins into cells is promoted by adenovirus particles. Substantial levels of ongoing translation on preexisting cellular mRNAs still continue for several hours after eIF4G degradation. Treatment of control HeLa cells with hypertonic medium causes an inhibition of translation that is reversed u...

Novoa, Isabel; Carrasco, Luis

1999-01-01

310

Evidence that a triplex-forming oligodeoxyribonucleotide binds to the c-myc promoter in HeLa cells, thereby reducing c-myc mRNA levels.  

OpenAIRE

A synthetic 27-base-long oligodeoxyribonucleotide, termed PU1, has been shown to bind to duplex DNA to form a triplex at a single site within the human c-myc P1 promoter. PU1 has been administered to HeLa cells in culture to examine the feasibility of influencing transcription of the c-myc gene in vivo. It is shown that uptake of PU1 into the nucleus of HeLa cells is efficient and that the compound remains intact for at least 4 hr. In nuclei extracted from PU1-treated cells, inhibition of DNa...

Postel, E. H.; Flint, S. J.; Kessler, D. J.; Hogan, M. E.

1991-01-01

311

Optical parameters measurement for diagnostic and photodynamic therapy of human cervical adenocarcinoma (HeLa) cell line  

Science.gov (United States)

The purpose of this study was to investigate the optical properties, absorption coefficient (? a ) scattering coefficient (? s ) and refractive indices, (n) of HeLa cell line in a suspension of 2% minimum essential medium (MEM) at two different (632.8 and 532.0 nm) wave lengths of laser light. Optical properties were determined with Kubelka Munk Model (KMM) and refractive index measurement was made through minimum angle of deviation method (MAD). We reported ? a = 8.643 ± 0.187 and 2.348 ± 0.249 cm-1 and ? s = 5.609 ± 0.287 and 88.166 ± 2.833 cm-1 at 632.8 and 532.0 nm, respectively. Refractive index was found to be 1.332 and 1.312 at 632.8 nm and 532.0 nm, respectively. The discussed results provide a route of information for clinical diagnosis, therapeutic application and dosimetry studies in HeLa and other cell lines.

Rehman, A.; Firdous, S.; Nawaz, M.; Ahmad, M.

2011-11-01

312

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

OpenAIRE

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

Ramer, Robert; Merkord, Jutta; Rohde, Helga; Hinz, Burkhard

2010-01-01

313

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

314

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

OpenAIRE

Alkylation chemotherapy has been a long-standing treatment protocol for human neoplasia. N-methyl-N’–nitro-N-nitrosoguanidine (MNNG) is a direct-acting monofunctional alkylator. Temozolomide is a clinical chemotherapeutic equivalent requiring metabolic breakdown to the alkylating agent. Both chemicals have similar mechanistic efficacy against DNA mismatch repair proficient tumor cells that lack expression of methylguanine methyltransferase (MGMT). Clinically relevant concentrations of bot...

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

2009-01-01

315

Comparative study of uptake and washout of 99TcmN(NOEt)2 with 99Tcm-MIBI in human cervical carcinoma cell line Hela  

International Nuclear Information System (INIS)

Objective: To investigate the cellular kinetics of 99Tcm-bis (N-ethoxy-N-ethyl dithiocarbamato)-nitrido [N(NOEt)2] in human cervical carcinoma Hela cells and to compare it with the cellular kinetics of 99Tcm-methoxyisobutylisonitrile (MIBI) in similar conditions; to define the possible clinical value of 99TcmN(NOEt)2 in tumor imaging. Methods: According to the principle of radionuclide tracer technique, 99TcmN(NOEt)2 or 99Tcm-MIBI were incubated with Hela cells at 37 degree C or at 22 degree C, at several time points after incubation, the uptake and washout characteristics of radiotracers in Hela cells were investigated and compared. Results: The maximum uptake of 99TcmN(NOEt)2 in Hela cells was 46.15%, that of 99Tcm-MIBI was 12.60%(P99TcmN(NOEt)2 after 5 min incubation in Hela cells was 65% of total uptake, uptake of 99Tcm-MIBI was 50% of total uptake (P99TcmN(NOEt)2 was retained in the Hela cells, 56.67% of 99Tcm-MIBI was retained (P99Tcm-MIBI, the cellular kinetics of 99TcmN(NOEt)2 was not temsup>mN(NOEt)2 was not temperature-dependent (the cellular kinetics was similar at 37 degree C and at 22 degree C, P > 0.05). Conclusion: In vitro data suggest that 99TcmN(NOEt)2 may be a tracer better than 99Tcm-MIBI for tumor imaging; 99TcmN(NOEt)2 has potential application in clinical use

316

Nonhepatic Cell Lines HeLa and 293 Support Efficient Replication of the Hepatitis C Virus Genotype 2a Subgenomic Replicon  

OpenAIRE

The hepatitis C virus (HCV) genotype 2a subgenomic replicon can replicate in two human non-hepatocyte-derived cell lines, HeLa and 293, with in vitro-transcribed replicon RNA. Sequencing analysis revealed that mutations in HCV-derived regions were not essential for replication in these cells, as some clones displayed no mutations.

Kato, Takanobu; Date, Tomoko; Miyamoto, Michiko; Zhao, Zijiang; Mizokami, Masashi; Wakita, Takaji

2005-01-01

317

Prevalence of Escherichia coli strains with localized, diffuse, and aggregative adherence to HeLa cells in infants with diarrhea and matched controls.  

OpenAIRE

To determine the possible role of Escherichia coli strains with three different patterns of adherence to HeLa cells in causing diarrhea in infants in São Paulo, Brazil, we studied stool specimens from 100 infants up to 1 year of age with acute diarrheal illnesses and 100 age-matched control infants without recent diarrhea. E. coli with localized adherence to HeLa cells was much more common in patients (23%) than in controls (2%) (P less than 0.0001) and was detected more frequently than rota...

Gomes, T. A.; Blake, P. A.; Trabulsi, L. R.

1989-01-01

318

Design and synthesis of new chacones substituted with azide/triazole groups and analysis of their cytotoxicity towards HeLa cells.  

Science.gov (United States)

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 IC?? value of 13.03 ?M, similar to the value of cisplatin (7.37 ?M). PMID:22932214

da Silva, Graziele D; da Silva, Marina G; Souza, Estrela M P V E; Barison, Andersson; Simões, Sarah C; Varotti, Fernando P; Barbosa, Leandro A; Viana, Gustavo H R; Villar, José A F P

2012-01-01

319

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

Directory of Open Access Journals (Sweden)

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

320

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.  

OpenAIRE

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 hig...

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

1994-01-01

321

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

Directory of Open Access Journals (Sweden)

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

2009-03-01

322

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

OpenAIRE

The aim of the study was to evaluate the concentrations of a soluble form of APO-1/Fas antigen (sFas, CD95) and a soluble Ligand for APO-1/Fas antigen (sCD95L, sFasL) in supernatants from CaSki and HeLa cell line cultures after the incubation with All-trans-retinoic acid. HPV-16 and HPV18 - positive cell lines were cultivated with All-trans-retinoic acid in concentrations of 1 x 10(-6) M/L and 1 x 10(-8) M/L. The cultures were incubated for 24 hours. Control culture with 3 microl of dimethyl-...

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

2010-01-01

323

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

324

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

325

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

326

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

International Nuclear Information System (INIS)

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

327

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  

Directory of Open Access Journals (Sweden)

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

328

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

329

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

330

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

331

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

332

Induced accumulation of polyubiquitin gene transcripts in HeLa cells after UV-irradiation or TPA-treatment  

International Nuclear Information System (INIS)

There are three species of ubiquitin gene transcripts in HeLa cells, termed UbA (? 0.65 kb), UbB (? 1.1 kb) and UbC (? 2.5 kb). In the present report, the UbC transcript was shown to be induced up to 2.5-fold after irradiation with UV light or treatment with phorbol ester (TPA). The kinetic analysis indicated the induction of the UbC gene was rapid and transient, as the maximal accumulation of the UbC transcript was induced at 2.5 h after UV irradiation or 3 h after TPA treatment. On the other hand, induction of UbA and UbB, in almost cases, was not observed. (author)

333

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 exposure ...

334

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

335

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

International Nuclear Information System (INIS)

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

336

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

International Nuclear Information System (INIS)

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

337

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

Directory of Open Access Journals (Sweden)

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

338

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 Cells were cultured as sph...

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

2012-01-01

339

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

340

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 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

341

Effect of inhibitors of poly(ADP-ribose)polymerase on the radiation response of HeLa S3 cells  

International Nuclear Information System (INIS)

The purpose of this study was to investigate possible involvement of poly(ADP-ribosyl)ation reactions in X-ray-induced cell killing, repair of potentially lethal damage (PLD), and formation and repair of radiation-induced DNA damage. As tools we used the inhibitors of poly(ADP-ribose)polymerase, 3-aminobenzamide (3AB), and 4-aminobenzamide (4AB). Both drugs inhibited PLD repair equally well but did not increase radiation-induced cell killing when cells were plated immediately after irradiation. 3AB affected repair of radiation-induced DNA damage, while 4AB had no effect. When 3AB was combined with aphidicolin (APC), it was found that the amount of DNA damage increased during the postirradiation incubation period. This means that the presence of 3AB stimulates the formation of DNA damage after X-irradiation. It is concluded that 3AB and 4AB sensitize HeLaS3 cells for radiation-induced cell killing by inhibiting repair of PLD. Because of the different effects of both inhibitors on repair of PLD and repair of radiation-induced DNA damage (a process known to be affected by inhibition of poly(ADP-ribosyl)ation), it is concluded that the observed inhibition of PLD repair is not caused by inhibition of poly(ADP-ribose)polymerase, and that the inhibitors affect repair of PLD and repair of DNA damage through independent mechanisms

342

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

343

Cancer Stem Cells  

OpenAIRE

Cancer stem cell theory gains increasingly greater significance in the world of medicine. Numerous findings of scientific research in vivo and in vitro indicate that it is the population of undifferentiated, self-renewing cells which is responsible for recurrence of cancer and metastasis. Similarly to normal stem cells, cancer stem cells (CSC) function in the environment of the other cells of the organism, called the niche, where they receive signals for differentiation and proliferation proc...

Aurelio Lorico; Eric Deutsch; Bo Lu; Shih-Hwa Chiou

2012-01-01

344

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

International Nuclear Information System (INIS)

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

345

RNA capping by HeLa cell RNA guanylyltransferase. Characterization of a covalent protein-guanylate intermediate.  

Science.gov (United States)

RNA capping by partially purified HeLa cell GTP:RNA guanylyltransferase has been shown to occur in the following sequence of two partial reactions involving a covalent protein-guanylate intermediate: (i) E(P68) + GTP in equilibrium E(P68-GMP) + PPi (ii) E(P68-GMP) + ppRNA in equilibrium GpppRNA + E(P68) Initially, the enzyme reacts with GTP in the absence of an RNA cap acceptor to form a covalent protein-guanylate complex. This complex consists of a GMP residue linked via a phosphoamide bond to a Mr = 68,000 protein. The enzyme then transfers the guanylate residue from the Mr = 68,000 polypeptide to the 5' end of diphosphate-terminated poly(a) to yield the capped derivative GpppA(pA)n. Both partial reactions have been shown to be reversible. In the reverse of Reaction i, E(P68--GMP) reacts with PPi to regenerate GTP. In the reverse of Reaction ii, the enzyme catalyzes the transfer of the 5'-GMP from capped RNA to the Mr = 68,000 protein to form protein-guanylate complex. A divalent cation is required for both partial reactions. The Mr = 68,000 protein is presumed to be a subunit of the HeLa guanylyltransferase. This interpretation is consistent with the sedimentation coefficient of 4.2 S of the native enzyme. Preliminary studies of RNA guanylyltransferase from mouse myeloma tumors suggest a similar mechanism of transguanylylation involving a Mr = 68,000 protein-guanylate complex. These data, in conjunction with previous studies of vaccinia virus guanylyltransferase (Shuman, S., and Hurwitz, J. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 187-191) suggests that covalent GMP-enzyme intermediates may be a general feature of the RNA capping reaction. PMID:6282835

Shuman, S

1982-06-25

346

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

OpenAIRE

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

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

2013-01-01

347

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

348

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

349

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)

350

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

Directory of Open Access Journals (Sweden)

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

351

Multidentate Small-Molecule Inhibitors of Vaccinia H1-related (VHR) Phosphatase Decrease Proliferation of Cervix Cancer Cells  

OpenAIRE

Loss of VHR phosphatase causes cell cycle arrest in HeLa carcinoma cells, suggesting that VHR inhibition may be a useful approach to halt the growth of cancer cells. We recently reported that VHR is upregulated in several cervix cancer cell lines as well as in carcinomas of the uterine cervix. Here we report the development of multidentate small-molecule inhibitors of VHR that inhibit its enzymatic activity at nanomolar concentrations and exhibit antiproliferative effects on cervix cancer cel...

Wu, Shuangding; Vossius, Sofie; Rahmouni, Souad; Miletic, Ana V.; Vang, Torkel; Vazquez-rodriguez, Jesus; Cerignoli, Fabio; Arimura, Yutaka; Williams, Scott; Hayes, Tikva; Moutschen, Michel; Vasile, Stefan; Pellecchia, Maurizio; Mustelin, Tomas; Tautz, Lutz

2009-01-01

352

The relationship between cell killing effect and number of lanthanide atoms bound to DNA molecules in cultured HeLa cells treated with rare earth elements  

International Nuclear Information System (INIS)

HeLa S-3 cells were treated with several rare earth elements, Ce, La, and Sm, at 37 deg C for 1 hour and the relationship between the lethal effects of these elements and the number of the atoms bound to DNA, RNA, and proteins was examined. Among the 3 elements, only Ce gave an exponential dose-survival relationship from which the value of mean lethal concentration (D0) was determined to be 6.75 mM. By using identically treated cells, the number of Ce atoms combined with DNA, RNA and protein molecules in HeLa cells were determined after fractionation of the cells using neutron activation analysis. In this way, the D0 value given as the Ce concentration was replaced by the number of Ce atoms combined with each fraction, then the target volumes, viz., cell killing efficiency, expressed as the reciprocal of D0 value was calculated for each fraction. The results suggested that DNA and RNA could stochiometrically be the primary target molecule for cell killing by Ce. (author)

353

Adenovirus proteins associated with mRNA and hnRNA in infected HeLa cells  

International Nuclear Information System (INIS)

The proteins that interact with cytoplasmic and nuclear polyadenylated RNA in adenovirus type 5 (Ad5) infection of HeLa cells were examined by UV-induced RNA-protein cross-linking in intact cells. The Ad5 100-kilodalton late nonvirion protein (100K protein) was cross-linked to both host and viral polyadenylated cytoplasmic RNA (mRNA). The cross-linking of the 100K protein to mRNA appears to correlate with productive infection, because the protein is not cross-linked to mRNA in abortive infection of wild-type Ad5 in monkey cells (CV-1) even though normal amounts of it are produced. However, when CV-1 cells are infected with Ad5 hr404, and Ad5 mutant which overcomes the host restriction to wild-type Ad5 infection in these cells, the 100K protein is cross-linked to mRNA. To identify and obtain antibodies to RNA-contacting proteins, a mouse was immunized with oligo(dT)-selected cross-linked RNA-protein complexes from Ad5-infected cells and the serum was used for immunoblotting experiments. It was found that in addition to the 100K protein, the Ad5 72K DNA-binding protein is also associated with RNA in the infected cells. The 72K DNA-binding protein is cross-linked to polyadenylated nuclear RNA sequences. These findings indicate that adenovirus proteins interact with RNAs in the infected cell and suggest possible mechanisms for the effects of the virus on mRNA metabolism

354

A CdtA-CdtC complex can block killing of HeLa cells by Haemophilus ducreyi cytolethal distending toxin.  

Science.gov (United States)

The cytolethal distending toxin (CDT) of Haemophilus ducreyi is comprised of the CdtA, CdtB, and CdtC proteins, with the CdtB protein having demonstrated enzymatic (i.e., DNase) activity. Using a single recombinant Escherichia coli strain with two plasmids individually containing the H. ducreyi cdtA and cdtC genes, we purified a noncovalent CdtA-CdtC complex. Incubation of this CdtA-CdtC complex with HeLa cells blocked killing of these cells by CDT holotoxin. Furthermore, the addition of purified recombinant CdtB to HeLa cells preincubated with the CdtA-CdtC complex resulted in the killing of these human epithelial cells. PMID:14573688

Deng, Kaiping; Hansen, Eric J

2003-11-01

355

Characterization of Chrysin Glucuronidation in UGT1A1-Overexpressing HeLa Cells: Elucidating the Transporters Responsible for Efflux of Glucuronide.  

Science.gov (United States)

Active transport of glucuronide out of cells is a critical process in elimination of drugs via the glucuronidation pathway. Here, HeLa cells were stably transfected with UGT1A1 and the contributions of BCRP and MRP family transporters to the cellular efflux of chrysin glucuronide (CG) were determined. The cDNA of UGT1A1 was introduced into HeLa cells using the lentiviral transfection method. The modified cells were functional in generation of the glucuronide from chrysin. Ko143 at 10-20 ?M (a dual inhibitor of BCRP and UGT1A1) caused a marked decrease (51.3%-59.7%, P resulted in a significant reduction in the excretion rate (18.2%-64.0%, P results suggested that BCRP, MRP1, MRP3, and MRP4 were significant contributors to excretion of CG. PMID:25595598

Quan, Enxi; Wang, Huailing; Dong, Dong; Zhang, Xingwang; Wu, Baojian

2015-04-01

356

Direct characterization of influenza viral NS1 mRNA and related sequences from infected HeLa cells and a cell-free transcription system.  

Science.gov (United States)

The NS1 mRNA of the influenza A virus WSN (H0N1) strain was isolated from a cell-free transcription system, and from the cytoplasm of virus-infected HeLa cells. The 32P-labeled NS1 mRNA derived from the infected cell cytoplasm was characterized by the secondary enzymatic analysis of sixteen of its large or distinct RNAase T1-resistant oligonucleotides. Several WSN strain-specific nucleotide differences from the previously-determined sequence of NS1 mRNA from the PR8 (H0N1) strain of influenza A virus, were located within these sequences. The RNAase T1-resistant oligonucleotides were placed within the primary sequence of NS1 mRNA, using the PR8 strain sequence data. The resulting linear map was then used to identify NS2 mRNA isolated from the infected cell cytoplasm, and an NS-related RNA species generated from NS1 mRNA incubated in a HeLa cell-free extract. PMID:3094582

Nicholson, A W; Frankfort, H M; Davis, N G; Ferrari, S; Lamb, R A; Robertson, H D

1986-11-13

357

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

Directory of Open Access Journals (Sweden)

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

358

Compatibility of cancer cells with nanostructured oxidized porous silicon substrates  

Energy Technology Data Exchange (ETDEWEB)

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

359

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

Scientific Electronic Library Online (English)

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

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

1036-10-01

360

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

OpenAIRE

Milky sap isolated from Chelidonium majus L. (Greater Celandine) serves as a rich source of various biologically active substances such as alkaloids, flavonoids and phenolic acids. Previous research showed that the activity of Ch. majus milky sap may depend also on the presence of biologically active proteins. The goal of this study was to evaluate the biological effect of two nucleases isolated from Ch. majus milk sap, CMN1 of 20 kDa and CMN2 of 36 kDa, on HeLa and CHO tumour cell lines. Bot...

Maria Wo?u?-Cholewa; Robert Nawrot; Anna Go?dzicka-Józefiak

2008-01-01

361

pEgr-sTRAIL transfer in combination with 60Co ? ray irradiation to induce the apoptosis on HeLa cells and activation of Caspase-3  

International Nuclear Information System (INIS)

In order to approach the radiosensitivity of TRAIL expression controlled by Egr-1 promotor, the recombinant vector pEgr-sTRAIL was tranfected into HeLa cells, the early apoptosis and Caspase-3 activity were detected after different doses of 60Co ?-ray irradiation. The results showed that pEgr-sTRAIL transfected in combination with ?-ray irradiation could significantly induce the apoptosis of HeLa cells in a dose-dependent manner. The higher activity of Capase-3 was also found in pEgr-sTRAIL irradiated group by using western blotting and spectrophotometry. Our result demonstrated that the activity of Caspase-3, as the apoptosis executor, play an important role in TRAIL-induced apoptosis and the enhanced TRAIL-induced apoptosis in transfected cells after irradiation can be controlled by radio-sensitive promoter Egr-1. (authors)

362

Gastric Cancer Stem Cells  

OpenAIRE

Cancer stem cells are defined as the unique subpopulation in the tumors that possess the ability to initiate tumor growth and sustain self-renewal as well as metastatic potential. Accumulating evidence in recent years strongly indicate the existence of cancer stem cells in solid tumors of a wide variety of organs. In this review, we will discuss the possible existence of a gastric cancer stem cell. Our recent data suggest that a subpopulation with a defined marker shows spheroid colony format...

Takaishi, Shigeo; Okumura, Tomoyuki; Wang, Timothy C.

2008-01-01

363

Liver Cancer Stem Cells  

OpenAIRE

Hepatocellular carcinoma is the most common primary malignancy of the liver in adults. It is also the fifth most common solid cancer worldwide and the third leading cause of cancer-related death. Recent research supports that liver cancer is a disease of adult stem cells. From the models of experimental hepatocarcinogenesis, there may be at least three distinct cell lineages with progenitor properties susceptible to neoplastic transformation. Identification of specific cell surface markers fo...

Mikhail, Sameh; He, Aiwu Ruth

2008-01-01

364

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

365

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

366

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

367

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.

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

2013-02-01

368

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

International Nuclear Information System (INIS)

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

369

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.

370

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

371

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

372

Suppression of the NF-kappaB signalling pathway by ergolide, sesquiterpene lactone, in HeLa cells.  

Science.gov (United States)

We have previously reported that ergolide, a sesquiterpene lactone isolated from Inula britannica, suppresses inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression by inhibiting nuclear factor-kappaB (NF-kappaB) in RAW 264.7 macrophages. In this study, we show that ergolide suppresses the DNA binding activity of NF-kappaB and nuclear translocation of NF-kappaB p65 subunit, leading to the inhibition of NF-kappaB-dependent gene transcription in 12-O-tetradecanoylphorbol 13acetate (TPA)-stimulated HeLa cells. We also show that ergolide decreases the degradation and phosphorylation of IkappaB, an inhibitory protein of NF-kappaB, and this effect is accompanied by a simultaneous reduction of IkappaB kinase (IKK) activity. However, ergolide does not inhibit in-vitro IKK activity directly, suggesting the possible involvement of upstream IKK kinases in the regulation of NF-kappaB activation. Furthermore, ergolide-mediated protein kinase Calpha (PKCalpha) inhibition is involved in reduction of NF-kappaB inhibition, as demonstrated by the observation that dominant negative PKCalpha, but not p44/42 MAPK and p38 MAPK, inhibits TPA-stimulated reporter gene expression. Taken together, our results suggest that ergolide suppresses NF-kappaB activation through the inhibition of PKCalpha-IKK activity, providing insight for PKCalpha as a molecular target for anti-inflammatory drugs. PMID:17430640

Chun, Jae Kwang; Seo, Dong-Wan; Ahn, Seong Hoon; Park, Jae Hyun; You, Jueng-Soo; Lee, Chang-Hee; Lee, Jae Cheol; Kim, Yong Kee; Han, Jeung-Whan

2007-04-01

373

Synthesis, analysis and cytotoxic evaluation of some hydroxypyridinone derivatives on HeLa and K562 cell lines.  

Science.gov (United States)

A range of iron bidentae ligands containing the chelating moiety 3- hydroxypyridin-4-ones (HPOs) have been synthesized via a single or a three-step synthetic pathway. In the single-step reaction, maltol was directly reacted by suitable primary amine and in the second synthetic method; benzylated maltol was reacted with related amines to give 1-substuted-2-methyl-3-benzyloxypyridin-4-one derivatives. Finally, removal of the benzyl group under acidic conditions was performed by catalytic hydrogenation to yield the favored bidentate chelators as HCl salt. The partition coefficient of the free ligands and their iron (III) complexes between an aqueous phase buffered at pH 7.4 and 1-octanol were also determined. The cytotoxic effects of these iron chelators against HeLa and K562 cell lines were evaluated using MTT assay and the results showed that cytotoxicity was closely related to the lipophilicity of compounds so that the most lipophilic compound (4g) revealed the highest activity and compound 4e as a more hydrophilic agent (Kpart; 0.05) showed the lowest cytotoxic effect. PMID:24019828

Saghaie, L; Sadeghi-Aliabadi, H; Ashaehshoar, M

2013-07-01

374

A novel chemosensor with visible light excitability for sensing Zn2+ in physiological medium and in HeLa cells.  

Science.gov (United States)

In the present study a novel imine-hydrazone based fluorescent chemosensor () for efficient and selective sensing of Zn(2+) over other biologically important metal ions under physiological conditions is reported. An enhancement in fluorescence emission intensity of the developed probe with a red shift of ?25 nm was observed for Zn(2+), whereas other metal ions failed to reveal any significant change in the emission spectra. Interestingly, the receptor functioned under completely physiological conditions (99.7% HEPES buffer) and has visible light excitability. Sensing of Zn(2+) was investigated in detail by absorption spectroscopy, emission spectroscopy, DFT calculation, (1)H-NMR titration experiment and ESI-MS experiment. The association constant between and Zn(2+) was found to be 5.58 × 10(5) M(-1). The receptor could detect as low as 69 ppb Zn(2+). Sensing of Zn(2+) is proposed through switch-on of intramolecular charge transfer (ICT) and chelation enhanced fluorescence (CHEF) processes after the introduction of Zn(2+) into the free ligand. The developed receptor was non-toxic and rendered intracellular sensing of Zn(2+) in HeLa cells through fluorescence imaging studies. PMID:24879606

Datta, Barun Kumar; Thiyagarajan, Durairaj; Samanta, Soham; Ramesh, Aiyagari; Das, Gopal

2014-07-21

375

Upregulation of endogenous farnesyl diphosphate synthase overcomes the inhibitory effect of bisphosphonate on protein prenylation in Hela cells.  

Science.gov (United States)

Nitrogen-containing bisphosphonates (N-BPs) such as zoledronic acid (ZOL) are the gold standard treatment for diseases of excessive bone resorption. N-BPs inactivate osteoclasts via inhibition of farnesyl diphosphate synthase (FPPS), thereby preventing the prenylation of essential small GTPases. Not all patients respond to N-BP therapy to the same extent, and some patients, for example with tumour-associated bone disease or Paget's disease, appear to develop resistance to N-BPs. The extent to which upregulation of FPPS might contribute to these phenomena is not clear. Using quantitative PCR and western blot analysis we show that levels of FPPS mRNA and protein can be upregulated in HeLa cells by culturing in lipoprotein deficient serum (LDS) or by over-expression of SREBP-1a. Upregulated, endogenous FPPS was predominantly localised to the cytosol and did not co-localise with peroxisomal or mitochondrial markers. Upregulation of endogenous FPPS conferred resistance to the inhibitory effect of low concentrations of ZOL on the prenylation of the small GTPase Rap1a. These observations suggest that an increase in the expression of endogenous FPPS could confer at least partial resistance to the pharmacological effect of N-BP drugs such as ZOL in vivo. PMID:24369118

Das, Subhajit; Edwards, Peter A; Crockett, Julie C; Rogers, Michael J

2014-04-01

376

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

International Nuclear Information System (INIS)

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

377

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

International Nuclear Information System (INIS)

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

378

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

379

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

380

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

OpenAIRE

Heat Shock Proteins 60 & 70 (HSP60 & HSP70) are intracellular protein that has been shown to be present at elevated levels in systemic circulation in Type 2 Diabetes mellitus (T2DM) patients. Conditions that lead to its secretion, and the mechanism of its translocation from cells, have not yet been defined. The aim of this study was to determine if specific cell stressors associated with T2DM, namely hyperglycaemia and oxidative stress, result in the upregulation of HSP60 in human cells in vi...

Hall, Luke; Martinus, Ryan D.

2013-01-01

381

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.)

382

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

OpenAIRE

FOXL2 is a transcription factor that is essential for ovarian function and maintenance, the germline mutations of which give rise to the blepharophimosis ptosis epicanthus inversus syndrome (BPES), often associated with premature ovarian failure. Recently, its mutations have been found in ovarian granulosa cell tumors (OGCTs). In this study, we measured the expression of FOXL2 in cervical cancer by immunohistochemistry and its mRNA level in cervical cancer cell lines Hela and Siha by RT-PCR. ...

Liu, Xing-long; Meng, Yu-han; Wang, Jian-li; Yang, Biao-bing; Zhang, Fan; Tang, Sheng-jian

2014-01-01

383

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

OpenAIRE

We report Static Magnetic Field (SMF) induced altered sub-cellular streaming, which retains even after withdrawal of the field. The observation is statistically validated by differential fluorescence recovery after photo-bleaching (FRAP) studies in presence and absence of SMF, recovery rate being higher in presence of SMF. This instant magneto-sensing by live cells can be explained by inherent diamagnetic susceptibility of cells and alternatively by spin recombination, e.g.,...

Raja, Sufi O.; Dasgupta, Anjan Kr

2014-01-01

384

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

385

Azithromycin synergistically enhances anti-proliferative activity of vincristine in cervical and gastric cancer cells.  

Science.gov (United States)

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-cancer effect on cancer over transformed cells in vitro. These results imply that AZM may be a potential anticancer agent for use in chemotherapy regimens, and it may minimize side effects via reduction of dosage and enhancing the effectiveness common chemotherapeutic drugs. PMID:24213508

Zhou, Xuezhang; Zhang, Yuyan; Li, Yong; Hao, Xiujing; Liu, Xiaoming; Wang, Yujiong

2012-01-01

386

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

Directory of Open Access Journals (Sweden)

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-cancer effect on cancer over transformed cells in vitro. These results imply that AZM may be a potential anticancer agent for use in chemotherapy regimens, and it may minimize side effects via reduction of dosage and enhancing the effectiveness common chemotherapeutic drugs.

Yujiong Wang

2012-12-01

387

Visualization and characterization of cancer stem-like cells in cervical cancer.  

Science.gov (United States)

Cancer stem cells (CSCs), defined by their differentiation capacity, self-renewal capacity, and maintenance of proliferation, have been identified in many tumors, including cervical cancer. Current studies identify CSCs by several specific biomarkers; however, it is difficult to monitor cervical CSCs in real-time in vitro and in vivo. Recent research reported the visualization of CSCs in breast cancer and gliomas using green fluorescent protein, ZsGreen, fused to a degron motif ornithine decarboxylase (ODC), which is destroyed by proteasomes. Accordingly, CSCs have low 26S proteasome activity, whereas non-CSCs have high 26S proteasome activity. Therefore, it is possible to observe CSCs by their accumulation of the fluorescent ZsGreen protein. In this study, we investigated optical imaging parameters to evaluate CSCs using two human cervical cancer cell lines: CaSki and HeLa. We defined populations as cell types having high- and low ZsGreen-cODC (high- and low-Zs, respectively) expression levels. The results of a sphere-forming assay revealed that the self-renewal ability of the high-Zs population was significantly higher than that of the low-Zs population. A tumorigenicity assay confirmed that the high-Zs population exhibited higher tumorigenic potential than the low-Zs population. The radioresistance of the high-Zs population of both HeLa and CaSki cells and the chemoresistance of the high-Zs population of CaSki cells were confirmed by a clonogenic survival assay and the tetrazolium dye assay, respectively. These results indicate that high-Zs populations of both the HeLa and CaSki cell lines possess CSC-like properties and therapeutic resistance. In conclusion, we successfully visualized CSC-like cells using a fluorescent protein system. PMID:25269542

Hayashi, Kazuhiko; Tamari, Keisuke; Ishii, Hideshi; Konno, Masamitsu; Nishida, Naohiro; Kawamoto, Koichi; Koseki, Jun; Fukusumi, Takahito; Kano, Yoshihiro; Nishikawa, Shimpei; Miyo, Masaaki; Noguchi, Kozo; Ogawa, Hisataka; Hamabe, Atsushi; Seo, Yuji; Doki, Yuichiro; Mori, Masaki; Ogawa, Kazuhiko

2014-12-01

388

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  

Directory of Open Access Journals (Sweden)

Full Text Available 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 test 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 6