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目的:体外研究5-氟尿嘧啶(5-fluororacil,5-Fu)联合米非司酮(mifepristone,MIF)对人宫颈癌SiHa细胞株的增殖抑制作用及可能的作用机制。方法:体外培养人宫颈鳞癌SiHa细胞株,采用MTT法检测不同浓度MIF、5-Fu单药以及两药联合作用对SiHa细胞增殖率的影响;采用FCM分析两药合用对SiHa细胞凋亡率及细胞周期分布影响;蛋白质印迹法检测药物处理后SiHa细胞中p53、Bax和Bcl-2蛋白表达的变化。结果:不同浓度5-Fu和MIF单药对SiHa细胞增殖均有抑制作用(P<0.05),且呈浓度和时间依赖性。5-Fu联合MIF作用于SiHa细胞可增强5-Fu对SiHa细胞的增殖抑制(P<0.05),且对MIF的作用浓度呈剂量依赖作用;蛋白质印迹法检测结果显示,联合用药组能使p53和Bax蛋白表达上调,Bcl-2蛋白表达下调,呈浓度依赖性。结论:MIF能增强5-Fu对SiHa细胞的增殖抑制和诱导凋亡作用,其机制可能与上调p53和Bax蛋白,下调Bcl-2蛋白表达有关。
OBJECTIVE: To study the inhibitory effect of 5-fluorouracil (5-Fu) and mifepristone (MIF) on the proliferation of human cervical carcinoma SiHa cell line in vitro and its possible mechanism. Methods: The human cervical squamous cell carcinoma SiHa cell line was cultured in vitro. MTT assay was used to detect the effect of different concentrations of MIF and 5-Fu on the proliferation of SiHa cells. FCM was used to analyze the apoptosis rate of SiHa cells And cell cycle distribution. Western blotting was used to detect the protein expression of p53, Bax and Bcl-2 in SiHa cells after drug treatment. Results: Different concentrations of 5-Fu and MIF alone inhibited the proliferation of SiHa cells (P <0.05) in a concentration-and time-dependent manner. The effect of 5-Fu combined with MIF on SiHa cells enhanced the inhibition of proliferation of SiHa cells by 5-Fu (P <0.05), and the dose-dependent effect of MIF was observed. Western blotting results showed that p53 And Bax protein expression, Bcl-2 protein expression was down-regulated, in a concentration-dependent manner. Conclusion: MIF can enhance 5-Fu on SiHa cell proliferation and apoptosis induced by the mechanism may be related to the up-regulation of p53 and Bax protein, down-regulation of Bcl-2 protein expression.