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Objective To investigate the biological features of epidermal growth factor receptor inhibitor AG1478 on cervical carcinoma cell. Methods The proliferation of HeLa cells under AG1478 stimulation was determined by CCK8 assay. The expression of EGFR downstream signaling protein and apoptotic relative protein were examined by Western blot and transcription of apotosis?related genes were measured by RT?qPCR in AG 1478 treated HeLa cells. Nuclear transport of phosphorylated ERK were measured through ICC assay. TUNEL assay was used to determine early stage of apoptosis. Results CCK8 assay showed that AG1478 inhibit the proliferation of HeLa cells and also block phosphorylated level of EGFR ,ERK and AKT. Furthermore ,nuclear transport of phosphorylated ERK upon EGF stimulation were blocked and pro?apoptotic proteins were up?regluated with activat ed cleaved Caspases. Conclusion AG1478 inhibited the proliferation and induced apoptosis in HeLa cells and it could be a potential therapeutic drug for cervical carcinoma.
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Objective To study the combined effect of interleukin-21 gene transfer and ionizing radiation on the growth of cervical carcinoma HeLa cells.Methods Previously constructed Ad-IL-21 gene was amplified by infecting 293A cells and the titer was measured by TCID50 method. HeLa cells were transfected with Ad-1L-21 and then irradiated with 6 Gy 137Cs γ-rays.The cells were divided into 5 groups,including blank control,Ad-LaeZ group,Ad-IL-21 group,radiation group and Ad-IL-21 combined with radiation group (combination group).The cell growth,cell cycle,apoptosis,and the expressions of IL-21 gene and protein in HeLa cells were detected.Results Ad-IL-21 was successfully amplified and the titer of Ad-11.-21 was 9 × 1010 pfu/ml.Compared with Ad-IL-21 group and radiation group,the cell growth of combination group was significantly inhibited at 96 h after transfection ( F =85.26,72.98,P ＜ 0.05 ).The cells in combination group were arrested in G1 phase and decreased at S phase( F =36.69,34.83,P ＜ 0.05),while the cellular apoptosis increased markedly ( F =28.23,25.57,P ＜ O.05 ). The gene expression of 1L-21 in the combination group was 1.54- and 2.43-fold of Ad-IL-21 group and blank control group,respectively (F=22.31,36.65, P ＜ 0.05 ), while the protein expression of IL-21 in the combination group was 1.62-fold and 2.31-fold of Ad-IL-21 group and blank control group,respectively ( F =27.36,35.86,P ＜ 0.05 ).Conclusions Ad-IL-21 gene transfection combined with radiation has synergic effect on the inhibition of cervical carcinoma cell growth.
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Objective:To explore the effect of inhibiting c-raf-1 by antisense technology on radiosensitivity in human cervical carcinoma cell line HeLa.Methods:There were four groups in the study:control group,lipofectin group,sense oligodexynucleotides(S-ODN) group,antisense oligodexynucleotides(AS-ODN) group.The expression of c-raf-1 was detected by means of RT-PCR,Cellular response to irradiation was evaluated by the colony forming test,Apoptosis was observed by fluorescent staining.Bcl-2 protein expression was determined by flow cytometry.Results:AS-ODN group could significantly decreased the proliferation rate and increasing the apoptosis rate,significantly downregulating the expression of Bcl-2 of irradiated human cervical carcinoma cells,(vs lipofectin group or S-ODN group,P
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OBJECTIVE: The purpose of this study is to determine the time-dependant effects of Glutathione on the Cisplatin-induced cytotoxicity of human cervical carcinoma cells. METHODS: Two human cervical carcinoma cells, SiHa (squamous cell carcinoma cell), and CaSki (epidermoid metastatic carcinoma cell) were cultivated with RPMI1640 media. Reduced glutathione (GSH) and 2-oxo 4-thiazolidine carboxylic acid (OTC) were added one hour before and after Cisplatin (2-50 micro M/ml) was applied. The cells were incubated an additional 24 hours and viable cells were examined using a 3-[4,5- dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. RESULTS: The MTT reduction rate of cisplatin-treated cervical carcinoma cells was increased significantly by the addition of glutathione (5 mM) or OTC (5 mM) both one hour before and after Cisplatin. A difference between MTT reduction rates one hour before and after cisplatin were not observed in either GSH or OTC. CONCLUSION: These results suggest that GSH and OTC have a protective effect on cisplatin-induced toxicity, and that this effect is about the same whether the agents were applied before or after the Cisplatin.
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Humans , Cisplatin , GlutathioneABSTRACT
Objective To investigate the mechanism of small-dose cisplatin in enhancing photodynamic therapy sensitivity and apoptosis in human cervical carcinoma cell lines.Methods The Hela and Siha cells were divided into four groups(blank control group,photodynamic therapy only,cisplatin only,photodynamic therapy and cisplatin).The effects on proliferation of Hela and Siha cells in the four groups were examined by MTT assay.The expression of P185c-erbB-2 in Hela and Siha cells affected was detected by immunocytochemistry.Results The degree of apoptosis caused by the joint application of cisplatin and photodynamic therapy was greater than that caused by cisplatin or photodynamic therapy alone.The expression of P185c-erbB-2 in cells declined correspondingly.However,the order of cisplatin and photodynamic therapy did not cause obvious differences in the effect.Conclusion The in vitro proliferation of Hela and Siha cells is restrained obviously by cisplatin combined with photodynamic therapy;the effect is greater than that of cisplatin or photodynamic therapy only.The mechanism of cispatin's enhancement of photodynamic therapy sensitivity may be related to inhibiting P185c-erbB-2 expression.
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OBJECTIVE: The purpose of this study is to determine the effects of glutathione on cisplatin-induced cytotoxicity of human cervical carcinoma cell lines (SiHa: squamous cell carcinoma cell, CaSki: epidermoid metastatic carcinoma cell). METHODS: Human cervical carcinoma cells (SiHa, CaSki) were incubated with culture media (RPMI1640) in the presence of cisplatin and/or buthionine sulfoximine (BSO), as a inhibitor of gamma-glutamyl- cysteine synthetase, and/or glutathione (GSH) and/or 2-oxo 4-thiazolidine carboxylic acid (OTC). The viable cells were examined by using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) and was determined by spectrophotometer at 570 nm. RESULTS: The incubation of cervical cells with cisplatin resulted in an decreasing cells viability by dose response. The MTT reduction rate were not different by BSO (5 mM) treatment in cervical cell lines. The viable cells were increased significantly by glutathione (5 mM) or OTC (5 mM) in cisplatin-treated cell lines. CONCLUSION: gamma-glutamylcysteine synthetase inhibitor had no effect on cisplatin toxicity. GSH and OTC had effect on cisplatin cytotoxicity. So, These result suggested that cervical cancer line cells were more susceptive to protective effects of glutathione and OTC than BSO on cisplatin induced-toxicity.
Subject(s)
Humans , Buthionine Sulfoximine , Carcinoma, Squamous Cell , Cell Line , Cisplatin , Culture Media , Cysteine , Glutathione , Ligases , Uterine Cervical NeoplasmsABSTRACT
Objective:To study the effects of celioscope carbon dioxide pneumoperitoneum on the growth of cervical carcinoma cell line in vitro. Methods:Cervical carcinoma cells CASKI were cultured in vitro,and then treated with 14mmHg,100% carbon dioxide for 2h. The proliferation,cell cycle and expressions of CDK4 and Cyclin D1 were examined by MTT, FCM and immunocytochemistry respectively. Results:After the treatment of carbon dioxide, the proliferation of cervical carcinoma cells CASKI was significantly increased at the 3th day, the proportion of proliferative phase in the cell cycle was significantly increased, and the expressions of CDK4 and Cyclin D1 were also significantly increased. Conclusion:carbon dioxide may promote the proliferation of cervical carcinoma cell line.
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Objective To investigate the correlation between methylation status and death-associated protein kinase 1 (DAPK1) in SiHa and Hela cell line of cervical carcinoma and intervention of DNA methyltransferase inhibitor 5-azacytidine on the expression of DAPK1 and the proliferation of the cells. Methods DAPK1 methylation status was analyzed using methylation-specific PCR methods. The expression of mRNA and protein of DAPK1 were analyzed by RT-PCR and SABC methods after the treatment with 5-azacytidine. MTT assay was used to observe the changes of proliferation activity of the cells after 5-azacytidine treatment. Results DAPK1 genes were methylated and did not express in SiHa cells in the cervical carcinoma. Its expression could be restored by 5-azacytidine. MTT assay showed 5-azacytidine could weaken the proliferation of cancerous cells. Conclusion DAPK1 methylation plays an important role in the carcinogenesis of cervical cells and can reexpress after the treatment with 5-azacytidine which also restored its inhibitory function on carcinoma.