靶向survivin的microRNA和siRNA抑制前列腺癌细胞survivin基因表达和凋亡作用的比较  

Comparison of inhibitory effects of microRNA and siRNA on survivin gene expression and apoptosis in prostate cancer

作　　者：骆晓梅[1] 刘家云[2] 苏明权[2] 郝晓柯[2] 

机构地区：[1]解放军第97医院检验科,江苏徐州221004 [2]第四军医大学西京医院全军临床检验医学中心,陕西西安710033

出　　处：《第四军医大学学报》2009年第4期331-335,共5页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30500433)

摘　　要：目的:构建靶向survivin的microRNA表达载体,观察其与siRNA表达载体对survivin基因表达的抑制作用.方法:以survivin为靶基因,根据pPRIME设计针对survivin的microRNA序列,获得目的片段克隆到相应载体中,序列正确的载体脂质体转染前列腺癌PC-3细胞,RT-PCR和Western Blot检测其对survivin基因表达的抑制作用,流式细胞术和电镜观察其对细胞凋亡的影响.结果:双酶切鉴定、DNA测序证实构建的microRNA表达载体序列正确,序列正确的microR-NA和siRNA表达载体在mRNA和蛋白水平上均能明显抑制PC-3细胞中survivin的表达,流式细胞术和电镜结果显示其可促进PC-3细胞的凋亡,且microRNA表达载体的作用效果较siRNA表达载体强.结论:Survivin靶向microRNA和siRNA均能抑制靶基因的表达,诱导前列腺癌细胞凋亡,与siRNA相比,microRNA作用效果更强.AIM ：To design and construct microRNA expression vectors, and to compare microRNA and siRNA inhibitive effects on survivin gene expression. METHODS：Based on survivin gene sequence, targeted pre-microRNA oligonucleotides were designed and synthesized respectively, and cloned into established microR-NA expression vector pPRIME. PC-3 cells were transiently trans-fected with correct constructs of microRNA and siRNA and the inhibitory effects were determined by PT-PCR and Western blot. The apoptosis index of PC-3 cells was detected by flow cytometry and electron microscope. RESULTS： Double enzyme digestion analysis and DNA sequencing results confirmed that the sequence of survivin specific microRNA expression vector was correct. Sur- vivin mRNA and protein expression level in PC-3 cells were specifically suppressed after transfection with recombinant plasmids. The results of flow cytometry indicated that the apoptosis index increased obviously in pPRIME-SV and pSilencer3.1-SW2 recom-binant plasmids. The results of cellular structure and cell organ in electron microscope displayed corresponding changes. The inhibitive effects of microRNA expression vector were better than those of siRNA expression vector. CONCLUSION： Both microRNA and siRNA expression vectors inhibit gene expression and induce apoptosis of prostate cells, but microRNA expression vector achieves better effect than siRNA expression vector.

关 键 词：靶向治疗 survivin 微RNAs RNA 小分子干扰 

分 类 号：R730.54[医药卫生—肿瘤]