RNA干扰survivin基因靶向治疗大肠癌

Effects and mechanisms of survivin-siRNA interference on colon adenocarcinoma cells SW620

目的:通过RNA干扰观察其对suivivin及PTEN在大肠腺癌中表达的差异及对大肠腺癌细胞增殖和凋亡的影响.方法:将survivin siRNA用脂质体2000转染SW620细胞,以荧光定量PCR仪检测survivin和PTEN mRNA的变化,Western蛋白印迹法半定量检测两者的蛋白变化,同时用MTT法检测细胞增殖,流式细胞仪检测细胞的凋亡和细胞周期分布的变化.结果:结肠腺癌SW620中免疫组化染色显示survivin呈阳性表达.siRNA作用于SW620细胞,survivin mRNA及蛋白的表达随着作用时间的延长而下调,PTEN mRNA及蛋白的表达随着作用时间的延长而呈现递增趋势,在12 h,24 h,48 h,survivin mRNA分别下调为对照组的75%,93.75%,97.8%,PTEN mRNA较对照组上调41%,100%,128%,均与对照组有显著性差异.MTT法检测siRNA干扰对细胞增殖的影响呈现出时间依赖性,与对照组相比有显著性差异(P<0.05),且作用各时间点之间比较亦有显著性差异(P<0.05).流式细胞仪检测细胞的凋亡率随时间的延长而增加.结论:survivin siRNA作用于结肠腺癌SW620可使survivin mRNA和蛋白的表达下调,同时使PTEN的表达上调,两者呈负相关,同时抑制细胞的增殖及促进细胞发生凋亡.

AIM： To investigate the effects and mechanisms of survivin-siRNA inteference on adenocarcinoma cells SW620. METHODS： SW620 cells were transfected with survivin-siRNA using Lipofectamine 2000. The mRNA expression levels of survivin and PTEN were detected by RT-PCR and protein expression levels were detected by Western blotting. MTT and flow cytometry were used to analyze proliferation and apoptosis. RESULTS： Compared with control ceils, the mRNA and protein levels of survivin were reduced in siRNA-transfected cells, while PTEN was increased, At 12, 24, 48 hours, the expression of survivin mRNA was downregulated 75%, 93.75% and 97.8%, respectivly, compared with that in the controls, and the expression of PTEN mRNA was upregulated 41%, 100%, 128%. The growth of transfected cells was inhibited, while apopotosis was increased. CONCLUSION： Survivin-siRNA has preferential effects on adenocarcinoma cells.