雷帕霉素增强Ishikawa细胞化疗敏感性实验研究

目的:研究雷帕霉素(RA)单独或联合阿霉素、顺铂及紫杉醇对子宫内膜癌细胞增殖和凋亡的影响,为临床应用RA治疗子宫内膜癌提供理论依据。方法:应用10μmol/L RA与各1/2的半数抑制浓度(IC50)量的阿霉素、顺铂及紫杉醇联合,细胞克隆形成法和流式法检测细胞存活率和凋亡率;RT-PCR法检测RA、阿霉素、顺铂及紫杉醇对细胞哺乳动物雷帕霉素靶蛋白(mTOR)mRNA表达影响;Western blot法检测10μmol/L RA作用6,12,24h后细胞Bcl-2蛋白表达水平。结果:RA联合化疗可显著提高各组化疗药的作用效果,降低细胞存活率,诱导细胞凋亡,与单纯化疗组相比有统计学差异(P<0.05);RA可显著降低宫颈癌细胞mTOR基因mRNA水平,抑制Bcl-2蛋白表达。结论:RA通过抑制mTOR通路,降低抗凋亡蛋白Bcl-2表达,提高化疗药物对子宫内膜癌细胞的杀伤作用。

Rapamycin Improves the Chemosensitivity of Ishikawa Cells in Vitro

Objective: To investigate the effect of Rapamycin(RA) on the proliferation apoptosis and drug sensitivity of human endometrial carcinoma line Ishikawa cells in vitro.Methods: Cell survival rate and cell apoptosis rate of Ishikawa cells were evaluated by clone formation test and flow cytometry after treatment of 10 μmol/L RA alone or combined respectively with 50% of half maximal inhibitory concentration(IC50) of the ADR,DDP,and PTX for 24 hours.The mRNA levels of mTOR after treatment was detected by RT-PCR.Western blot was used to measure Bcl-2 protein abundance after treatment with 10 μmol/L RA for 6,12,and 24 hours respectively.Results: Rapamycin significantly decreased the mRNA expressions of mTOR and the abundance of Bcl-2 protein.Compared with the treatment with RA alone,combined treatment significantly increased cell apoptosis rate,decreased cell survival rate and improved the cytotoxicity of the chemotherapeutic agents(P<0.05).Conclusion: Rapamycin can block the mTOR pathway and decrease Bcl-2 protein to inhibit the proliferation,induce the apoptosis and enhance the chemosensitivity of Ishikawa cells.