三氧化二砷在诱导非小细胞肺癌NCI-H157细胞凋亡过程中对Survivin基因表达Caspase-3的影响

The Impact of As_2O_3 on Expression of Survivin and Caspase-3 in the Process of Inducing NCI-H157 cell Apoptosis in Non-Small Cell Lung Cancer

目的:研究三氧化二砷(As2O3)在诱导非小细胞肺癌NCI-H157凋亡过程中对Survivin基因表达及Caspase-3的影响,探讨As2O3诱导肿瘤细胞凋亡的作用机理研究。方法:首先对NCI-H157细胞培养和增殖测定,然后采用RT-PCR法检测Survivin mRNA表达;Western Blot检测Survivin及Caspase-3蛋白。结果:RT-PCR结果显示,10μmol/L As2O3作用于NCI-H157细胞12h,24h,48h同对照组相比,Survivin mRNA表达分别降至85.7%,59.4%,22.5%,处理72h后几乎检测不到Survivin mRNA的表达。Western Blot解析结果显示,Survivin蛋白表达在5μmol/L As2O3处理后分别下调至75.2%(12h),54.3%(24h),37.5%(48h)及20.3%(72h)。分别用不同浓度的As2O35～20μmol/L作用NCI-H157细胞24h,Survivin蛋白表达随着As2O3浓度的增加逐渐下降。As2O3作用12h之内只能检测到非激活状态32KD的proCaspase-3,处理24h后出现17KD的活性亚基,直至72h。结论:As2O3以时间、剂量依赖的方式抑制NCI-H157细胞增殖,诱导细胞凋亡过程中下调Survivin mRNA及蛋白表达,并活化Caspases-3,提示As2O3诱导的NCI-H157细胞凋亡机制可能与下调Survivin基因表达及激活Caspase-3活性有关。

To examine the expression of survivin and casepase-3 in AszO3-induced apoptosis.and investigate the mechanisms of apoptosis-inducing effect of As2O3 on NCI-H157 cells. Methods： Survivin mRNA expression was detected by semi-quantitative RT-PCR. Survivin and caspase-3 were analyzed by Western blot. Results： Apoptosis of NCI-H157 cells was induced by As2O3 at concentrations of 10μmol/L and above. Compared with the control group, the survivin RNA expression rate in 101μmol/L As2O3 treatment group ＇after 12h, 24h and 48h was 85.7%, 59.4%, and 22.5%, respectively, and could hardly be detected after 72h. Western }）lot detection showed that the survivin protein expression rate was 75.2%（12h）, 54.3%（24h）, 37.5% （48h） and 20.3%（72h）, respectively. The cleaved active subunits of caspase-3 were observed within 24h to 72h after As2O3 treatment. Conclusion： As2O3 induced NCI-H157 cell apoptosis accompanied with survivin downr egulation and activation of caspase-3.