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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 investigate the effect and mechanism of mifepristone on the proliferation of human cervical squamous cell carcinoma Caski cells. To provide experimental evidence for the clinical application of mifepristone in the treatment of cervical squamous cell carcinoma. Methods: Human cervical squamous cell carcinoma Caski cells were cultured in vitro. Caski cells were treated with different concentrations of mifepristone. MTT assay was used to determine the effect of mifepristone on the proliferation of Caski cells The changes of cell cycle were analyzed by flow cytometry. Fluorescein isothiocyanate (FITC) fluorescence labeling flow cytometry (FCM) was used to detect the expression and activity of HPV-E6 and p53 in Caski cells treated with mifepristone. Results: ① MTT colorimetric assay showed that: Mifepristone at a concentration of 12.5 mg / L could inhibit the growth of Caski cells cultured in vitro in a concentration-time-dependent manner. Mifepristone 1.25 mg / L had no significant effect on Caski cells No significant inhibition (IR <5%). Flow cytometry showed that the concentration of 12.5 mg / L of mifepristone could block the Caski cell cycle in G1 phase in a concentration-time-dependent manner. ③ FITC fluorescence labeling FCM results showed that: After mifepristone on Caski cells, HPV-E6 protein expression was downregulated, p53 protein upregulation, in a concentration-dependent manner (P <0.05). Conclusion: (1) High dose (12.5-20 mg / L) of mifepristone can inhibit the growth of Caski cells and arrest the cell cycle in G1 phase. ② mifepristone can inhibit the proliferation of Caski cells, which is related to the down-regulation of HPV16-E6 protein expression and up-regulation of p53 protein expression.