米非司酮对宫颈癌Caski细胞增殖的影响

Effect of mifepristone on proliferation of cervical carcinoma Caski cells

目的:探讨米非司酮对人宫颈鳞癌Caski细胞增殖的影响和机制。为临床应用米非司酮治疗宫颈鳞癌提供实验依据。方法:体外培养人宫颈鳞癌Caski细胞,应用不同浓度的米非司酮处理Caski细胞,采用四甲基偶氮唑蓝(MTT)比色法,测定米非司酮对Caski细胞增殖活性的作用;流式细胞术分析细胞周期的变化;异硫氰酸荧光素(FITC)荧光标记流式细胞术(FCM)法,测定米非司酮对Caski细胞HPV-E6,p53的表达和活性变化。结果:①MTT比色法结果显示:12.5mg/L以上浓度米非司酮可抑制体外培养的Caski细胞生长,且呈明显的浓度-时间依赖方式,1.25mg/L的米非司酮对Caski细胞无显著的抑制作用(IR<5%)。②流式细胞术结果显示:12.5mg/L以上浓度的米非司酮能使Caski细胞周期阻滞于G1期,呈明显的浓度-时间依赖方式。③FITC荧光标记FCM法结果显示:米非司酮作用于Caski细胞后,HPV-E6蛋白表达下调,p53蛋白表达上调,呈浓度依赖方式(P<0.05)。结论:①大剂量(12.5～20mg/L)米非司酮具有抑制Caski细胞生长作用,能使其周期阻滞于G1期。②米非司酮对Caski细胞增殖有抑制作用,与下调HPV16-E6蛋白表达,上调p53蛋白表达有关。

Objective： To explore the effect of mifepristone on proliferation of human cervical carcinoma Caski cells and the mechanism, provide a experimental basis for treatment of cervical squamous carcinoma with mifepristone. Methods： Human cervical squamous carcinoma cell line Caski cells were cuhured in vitro, then Caski ceils were treated with various concentrations of mifepristone, the effect of mifepristone on proliferation of human cervical carcinoma Caski cells was detected by 4 methyl thiazolyl tetrazolium （MTT） assay; the changes of cell cycle was analyzed using flow cytometry ; the changes of expression levels and activity of HPV - E6 and p53 in Caski cells were investigated by FITC staining flow eytometry. Results： MTT assay indicated that mifepristone （more than 12.5 mg/L） could suppress the pro- liferation of Caski cells in vitro, showing a obvious concentration - time trend, 1.25 mg/L mifeprlstone had no significant effect on proliferation of Caski cells （IR 〈 5% ） . The results of flow cytometry indicated that mifepristone more than 12.5 mg/L could inhibit the cell cycle of Caski cells at G1 phase, showing a obvious concentration -time trend. FITC staining flow cytometry indicated that the expression of HPV16 - E6 protein decreased and the expression of p53 protein increased in Caski cells treated with mifepristone, showing a obvious concentration de- pendent trend （P 〈 0. 05） . Conclusion： Large dose of mifepfistone （12. 5 -20. 0 mg/L） can suppress the growth of human cervical squamous carcinoma Caski cells significantly, and arrest cell cycle at G1 phase. Mifepristone can suppress the proliferation of Caski cells, the mechanisms are related to down - regulation of HPV16 - E6 protein and up - regulation of p53 protein.