【摘 要】
：
目的 研究大黄素对人膀胱癌细胞BIU87凋亡及凋亡诱导因子(apoptosis inducing factor,AIF)和核酸内切酶G( Endonuclease G,Endo G)的mRNA和蛋白表达的影响,探讨大黄素的抗癌
【机 构】
：
宁波大学医学院附属宁波市第一医院泌尿外科
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目的 研究大黄素对人膀胱癌细胞BIU87凋亡及凋亡诱导因子(apoptosis inducing factor,AIF)和核酸内切酶G( Endonuclease G,Endo G)的mRNA和蛋白表达的影响,探讨大黄素的抗癌机制. 方法 取对数生长期的人膀胱癌细胞株BIU87,分别加入大黄素和Caspase抑制剂Z-VAD-FMK共同培养.实验分为4组:空白对照组、Z-VAD-FMK组、大黄素组、大黄素+Z-VAD-FMK组.采用甲基噻唑(MTT)比色法测定不同浓度大黄素在不同作用时间内对体外培养的BIU87细胞增殖的影响,应用流式细胞仪检测细胞凋亡,RT-PCR检测各组细胞AIF和Endo G mRNA的表达,蛋白质印迹法检测各组细胞AIF和Endo G蛋白的表达. 结果 MTT检测显示,大黄素的浓度越高、作用时间越长,对肿瘤细胞生长的抑制作用越强.根据IC50值,选择80 μmol/L的大黄素干预72 h作为干预条件.大黄素组细胞凋亡率为(44.57±1.52)％,大黄素+Z-VAD-FMK组为(35.58±1.61)％,差异有统计学意义(P＜0.05).RT-PCR和蛋白质印迹结果显示,大黄素+Z-VAD-FMK组中AIF mRNA和蛋白为1.74±0.11、2.59±0.13,大黄素组为1.36±0.08、1.89±0.14,空白对照组为0.37±0.02、0.53±0.11,Z-VAD-FMK组为0.42±0.06、0.44 ±0.07,大黄素+Z-VAD-FMK组与其他各组比较,差异有统计学意义(P＜0.05)；大黄素+ Z-VAD-FMK组中Endo G mRNA和蛋白为2.28±0.15、3.31±0.36,大黄素组为1.85±0.13、2.15 ±0.27,空白对照组为0.53±0.07、0.71 0.16,Z-VAD-FMK组为0.61±0.04、0.67±0.22,大黄素+Z-VAD-FMK组与其他各组比较,差异有统计学意义(P＜0.05). 结论 大黄素可上调膀胱癌细胞BIU87中AIF和Endo G的表达,通过启动非Caspase依赖途径诱导细胞凋亡.
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