Livin靶向小干扰RNA对前列腺癌PC3细胞生物学特性的影响  

Effect of siRNA targeting Livin gene on biological characteristics of human prostate cancer PC3 cells

作　　者：谢晓强[1] 章振保[1] 杨恩明[1] 李显文[1] 李宗金[2] 徐勇[3] 

机构地区：[1]厦门市第二医院泌尿外科,361021 [2]南开大学生物活性材料教育部重点实验室 [3]天津医科大学第二医院泌尿外科天津市泌尿外科研究所

出　　处：《中华泌尿外科杂志》2013年第11期859-863,共5页Chinese Journal of Urology

基　　金：福建省自然科学基金（2008-59-14）

摘　　要：目的 构建Livin靶向小干扰RNA（siRNA）重组表达载体,观察其对前列腺癌PC3细胞凋亡和增殖等生物学特性的影响. 方法 2009年1月至2012年1月,构建针对Livin基因的siRNA真核表达载体U6/GFP/Neo-Livin并转染前列腺癌PC3细胞.实时荧光定量PCR和蛋白质印迹法分别检测转染后Livin基因mRNA和蛋白的表达情况,MTT法检测对细胞增殖的影响,流式细胞仪检测转染后细胞的凋亡效应以及对细胞周期的影响,克隆形成实验观察对细胞体外克隆形成能力的改变,体内成瘤实验比较细胞在裸鼠体内成瘤性的变化. 结果 Livin靶向siRNA重组表达载体转染PC3细胞后,与对照组相比,实验组细胞内Livin mRNA和蛋白的表达水平分别下调至（31.55±3.92）％、（0.37±0.02）％（P＜0.01）；与对照组相比,PC3细胞增殖受到显著抑制（P＜0.01）；细胞凋亡率增到（26.5±3.3）％（P＜0.01）；Caspase3活性增至0.079±0.017 （P＜0.05）,体外克隆形成率降至（34.8±3.5）％（P＜0.01）.实验组和对照组荷瘤裸鼠的成瘤体积分别为（1.79±0.07）、（4.40±0.06） cm3（p＜0.01）. 结论 靶向Livin基因的siRNA干扰了前列腺癌PC3细胞中Livin基因的表达,抑制了细胞增殖并诱导其凋亡,延缓了肿瘤细胞的生长.Objective To observe the effect of RNAi targeting Livin gene on biology characteristics such as apoptosis and proliferation in human prostate cancer cells. Methods siRNA expression vector tar- geting Livin gene was constructed and transfected into human prostate cancer cell line PC3. The expressions of Livin mRNA and protein were detected by real-time PCR and Western-blot, cell apoptosis and cell cycle were assayed by flow cytometry, proliferation and colony formation were detected by MTT and colony forma- tion assay, and the tumor growth in vivo was observed in nude mice. Results After transfection, down- regulation of Livin mRNA and protein expression in PC3 cells was observed （P〈0.01）. Compared with the control group, the proliferation of cancer cells was inhibited significantly （P〈0.01） and the apoptotic ratio was （26.5±3.3）% （P〈0.01）. The Caspase3 activity increased obviously （P〈0.05） , and the experimental group showed a decreased colony formation rate （ P〈0.01 ）. The tumor volume of xenografts in nude mouse in experimental and control group was （ 1. 79 ± 0. 07 ） and （ 4.40 ± 0.06） cm3 respectively （ P 〈 0.01 ）. Conclusions The siRNA recombinant expression vector targeting Livin gene was constructed and can knockdown the expression of Livin mRNA and protein. It can inhibit PC3 cell proliferation, induce apoptosis and inhibit tumor growth in vivo.

关 键 词：凋亡抑制蛋白LIVIN RNA干扰 前列腺肿瘤 细胞凋亡 基因疗法 

分 类 号：R737.25[医药卫生—肿瘤]