靶向Survivin RNAi诱导人黏液表皮样癌细胞凋亡的实验研究

Study of apoptosis of human mucoepidermoid carcinoma cells induced by target Survivin RNAi

目的:应用RNA干扰技术靶向阻断Survivin基因表达,探讨其对黏液表皮样癌细胞的诱导凋亡作用。方法:将RNA干扰表达载体Pgenesil-sur通过脂质体介导转染人黏液表皮样癌细胞,采用琼脂糖凝胶电泳检测"梯状条带";流式细胞仪检测细胞凋亡率和凋亡峰。结果:转染48h后琼脂糖凝胶电泳可见180-200bp凋亡带;对照组、空载体组未见凋亡带。流式细胞仪检测细胞凋亡率结果显示,靶向SurvivinRNA干扰组24h、48h、72h分别为26.32%、34.56%、32.41%;与转染空载体(1.26%、1.28%、1.27%)和空白对照组(1.22%、1.25%、1.24%)比较显著增高(P<0.01);空载体组与空白对照组比较无统计学差异(P>0.05)。靶向SurvivinRNA干扰组出现凋亡峰。结论:靶向SurvivinRNA干扰能诱导体外培养的人黏液表皮样癌细胞发生凋亡。

Objective Target blocking the expression of Survivin gene by RNA interference skill to investigate its induction of apoptosis for human mucoepidermoid carcinoma cells. Metheds Transfect RNA interference expression vector to human mucoepidermoid carcinoma cells mediated by lipidosome, detect ＂ladder＂ by agarose gel electrophoresis, detect the apoptosis rate and apoptotic peak by flow cytometer. Results Apoptosis zone of 180- 200bp can be seen in agarose gel electrophoresis 48h after transfection; non apoptosis zone can be seen in the control group and pgenesil-1 group. The result of detect by flow cytometer showed the separate apoptosis rate in 24h,48h, 72h ： 26.32%,34.56%,32.41% for interference group; 1.26%, 1.28%,1.27% for Pgenesil-1 group; 1.22%,1.25%, 1.24% for control group; a significant increase can be seen between interference group and Pgenesil-1 group or control group （P〈0.01）; but there is no significant difference between pgenesil-1 group and control group （P〉0.05）. Apoptotic peakappeared in target RNA interference group. Conclusion Target survivin RNA intervention can induce the apoptosis of human mucoepidermoid carcinoma cells in vitro.