RNA干扰PLCε诱导人膀胱癌BIU-87细胞凋亡的实验研究  

RNAi PLCε INDUCES APOPTOSIS IN HUMAN BLADDER CANCER CELL LINES BIU-87

作　　者：赵懿[1] 罗春丽[1] 郭永灿[1] 欧俐苹[1] 

机构地区：[1]重庆医科大学医学检验系临床检验诊断学省部共建教育部重点实验室重庆市重点实验室,重庆400016

出　　处：《中国组织化学与细胞化学杂志》2010年第2期135-139,共5页Chinese Journal of Histochemistry and Cytochemistry

基　　金：重庆市教委2008年科学技术研究项目(A类)(KJ080306)

摘　　要：目的探讨以RNA干扰沉默人源磷脂酶Ce(phospholipase Cepsilon,PLCε)基因表达后诱导人膀胱癌细胞株BIU-87细胞凋亡的作用和机制。方法脂质体介导重组阳性质粒pGenesil-PLCε(以下简称P)和阴性质粒pGenesil-NP(以下简称NP)转染BIU-87细胞48h后,用RT-PCR和Western blot检测转染前后PLCεmRNA和蛋白表达,流式细胞术检测细胞周期和细胞凋亡率,电镜观察细胞形态改变。结果转染P质粒后可明显抑制PLCεmRNA和蛋白表达水平,抑制率分别为78.7%和76.6%;流式细胞术显示P质粒转染组细胞周期发生改变呈明显G0/G1期阻滞,并出现亚二倍体凋亡峰,细胞凋亡率增加(P<0.05);电镜观察P质粒转染组细胞可见凋亡小体。结论:RNA干扰沉默PLCε基因表达可诱导膀胱癌BIU-87细胞凋亡,其作用机制与细胞周期分布的改变有关。Objective To investigate the induced apoptotic effect of shRNA（short hairpin RNA）-mediated silence of PLCε gene expression on bladder cancer BIU-87 cells and its mechanism.Methods PLCε shRNA expression vector P and negative plasmid NP were transfected to BIU-87 cells by liposome.After 48 hours,RT-PCR and Western blot were used to detect PLCε expression in experimental and control groups.Flow cytometry was used to detect the cell cycle and cell apoptosis.Electron microscope was used to observe the morphological changes.Results The expression of PLCε mRNA and protein decreased in BIU-87 cells by the transfection of PLCε shRNA recombinant plasmid,and the inhibitory rates were 78.7% and 76.6%.Flow cytometric profiles revealed that positive plasmid transfection led to the increase of the cell percentage of G0/G1 phase and the cell percentage of apoptosis（P〈0.05）.Distinct morphological changes of cell apoptosis were observed by electron microscopy.Conclusion Transfection of PLCε shRNA can induce apoptosis of bladder cancer cells.The mechanism may be changing the tumor cell cycle.

关 键 词：TCCB 凋亡 PLCε SHRNA 

分 类 号：R737.11[医药卫生—肿瘤]