三氧化二砷诱导宫颈癌细胞凋亡与Fas/FasL表达的研究

Study on the Apoptosis of Cervical Cancer Cell by Arsenic Trioxide and the Expression of Fas/FasL

目的:研究三氧化砷(Arsenic trioxide,As2O3)体外诱导宫颈癌Hela和Si Ha细胞株凋亡作用及凋亡相关机制。方法:采用凝胶电泳、流式细胞术等方法进行凋亡检测,免疫组织化学方法检测As2O3诱导对宫颈癌Hela和Si Ha细胞株细胞Fas和FasL表达的影响。结果:2.5μmol/L和5μmol/L浓度的As2O3作用Si Ha和Hela细胞株48h可以观察到细胞出现典型的凋亡变化。琼脂糖凝胶电泳DNA片段观察到DNA梯形条带。流式细胞仪分析细胞凋亡率显示,As2O3诱导两种细胞株凋亡具有剂量和时间依赖性(P<0.05)。而且细胞免疫组化显示As2O3诱导两种细胞株凋亡同时上调Fas蛋白表达(P<0.05)。5μmol/L浓度的As2O3诱导Hela细胞株凋亡过程同时伴FasL蛋白表达明显上调(P<0.05)。结论:As2O3能诱导Si Ha和Hela细胞株凋亡,其机制可能与Fas基因的表达上调有关。

Objective：To study the mechanism of apoptosis on cervical cancer cell lines（Hela and SiHa）by As2O3.Methods：The DNA fragmentation analysis was determined by the gel electrophoresis.The cell apoptosis ration was measured by flow cytometry（FCM）.Cell immunohistochemical method was given to detect the expression of apoptosis-related protein Fas and Fas ligand.Results：We can observe the typical apoptotic cells in the concentrations of 2.5 μmol/L and 5 μmol/L As2O3 after 48 h treatment.DNA Ladder was seen.The result of FCM assay demonstrated：the apoptosis induced by As2O3 was time dose-dependent（P〈0.05）.As2O3（2.5 μmol/L,5 μmol/L）could significantly upregulate the expression of Fas on the two cell lines（P〈0.05）,5 μmol/L As2O3 also up regulated the expression of FasL on Hela cell lines（P〈0.05）.Conclusions：Apoptosis does occur on cervical carcinoma cell lines induced by As2O3.The upregulation of Fas maybe one of the main mechanisms of apoptosis on cervical cancer cell lines by As2O3.