survivin基因的RNA干涉抑制人乳腺癌SKBr-3细胞体外增殖的作用  

Inhibitory effect on human breast cancer SKBr-3 cells by RNA interference of survivin gene

作　　者：李庆霞[1] 黄红艳[1] 刘家云[2] 许彦鸣[1] 贾林涛[1] 赵晶[1] 张勇[1] 王成济[1] 杨安钢[1] 

机构地区：[1]第四军医大学生物化学与分子生物学教研室 [2]第四军医大学免疫学教研室,陕西西安710032

出　　处：《中国癌症杂志》2005年第2期101-104,共4页China Oncology

基　　金：国家自然科学基金(No.30200274);国家重点基础研究发展基金(No.2004CB518805)。

摘　　要：目的:研究RNA干涉(RNAinterference,RNAi)抑制survivin基因表达后对人乳腺癌SKBr3细胞体外增殖能力的影响。方法:通过RTPCR和间接免疫荧光检测转染survivin靶向的小干涉RNA表达载体pSUPERS1后SKBr3细胞中survivin基因的表达;集落形成实验、MTT比色法检测细胞体外增殖能力;流式细胞仪检测细胞周期。结果:转染pSUPERS1后,SKBr3细胞中survivin的mRNA和蛋白表达水平明显降低;克隆形成率(38±16.70)%比对照组(90.3±4.04)%显著降低;细胞增殖减慢,主要阻滞于G1期(74.03±8.91)%。结论:RNA干涉沉寂survivin基因表达后明显抑制了人乳腺癌SKBr3细胞的体外增殖能力。Purpose:To study the RNA interference-mediated inhibition of survivin gene on the proliferation of human breast cancer SKBr-3 cells. Methods:SKBr-3 cells were transfected with a pSUPER-S1 vector plasmid that expressed survivin-targeted small interfering RNA, and the mRNA and protein levels of survivin gene were measured with RT-PCR and indirect immunofluorescence, respectively. The proliferation of transfected SKBr-3 cells was investigated through colony forming assay and MTT assay. The cell cycle phases were determined by flow cytometry(FCM). Results:The mRNA and protein levels of survivin declined markedly in pSUPER-S1-transfected SKBr-3 cells. And the colony forming rate of those cells(38±16.70)% was significantly lower than that of the control cells(90.3±4.04)%.The growth of the pSUPER-S1-transfected cells was decelerated and the cell cycle was mainly blocked at G_(1) phase(74.03±8.91)%. Conclusions:survivin gene silencing by RNA interference contributed to a distinctive inhibition of the proliferation of human breast cancer SKBr-3 cells in vitro.

关 键 词：survivin RNA干涉 乳腺癌细胞 

分 类 号：R737.9[医药卫生—肿瘤]