RNAi沉默整合素连接激酶基因对人膀胱癌BIU-87细胞体内成瘤的影响  

RNA interference silences ILK gene and affects carcinoma tumorigenesis in vivo in Bladder Cancer BIU-87 cells

作　　者：何江[1] 陈亮[1] 王德林[1] 黄飚[1] 

机构地区：[1]重庆医科大学附属第一医院泌尿外科,重庆400016

出　　处：《激光杂志》2012年第2期69-70,共2页Laser Journal

基　　金：国家自然科学基金资助项目(30972999);重庆市自然科学基金资助项目(CSTC2005BB5033)

摘　　要：目的:研究RNAi沉默整合素连接激酶(integrin-linked kinase,ILK)基因对人膀胱癌BIU-87细胞裸鼠皮下成瘤的影响。方法:构建4条针对ILK基因的特异性miRNA干扰载体和1条阴性对照载体,并利用脂质体转染法将其转染BIU-87细胞,并筛选获得稳定转染细胞株。利用RT-PCR和Western blot分别从mRNA和蛋白水平检测抑制效果。将10只裸鼠随机分为2组,皮下分别接种转染组细胞和未转染组细胞,观察瘤体的生长情况,并于接种4周后处死裸鼠,测量其瘤体体积和重量,并将瘤体做HE染色,观察瘤体病理情况。结果:转染组细胞ILK的表达明显受到抑制,以miR-3组的抑制效率最高。裸鼠皮下成瘤实验检测发现:两组均有瘤体形成,未转染组较转染组瘤体生长快,未转染组体积平均为:289.56±36.49mm3,转染组为:56.67±4.32mm3。瘤体重量分别为:1.265±0.02 g和0.518±0.03g。转染组与未转染组相比差异具有统计学意义(P<0.05)。结论:利用RNAi技术能有效抑制靶基因ILK的表达,从而降低膀胱癌细胞的体内的增殖能力。Objective：To evaluate the inhibition effect of ILK gene silencing on tumor growth of human bladder cancer cell lines BIU-87 in vivo.Methods：Four specific miRNA RNAi vectors pcDNATM6.2-GW/EmGFP-ILK-miR（miR-1,miR-2,miR-3,miR-4） targeting human ILK and a negative control group were synthesized and transfected into the BIU-87 cells by liposome.The stable expression cell strain were obtained by continuing pressure screening of blasticidin and limiting dilution assay cultivation.The suppression effect targeting ILK was by Western-blot.10 nude mice were divided into 2 groups.One group subcutaneous implant transfected bladder cancer cells and another group subcutaneous implant non-transfected bladder cancer cells BIU-87.Morphological,histopathology and tumor inhibition rate were evaluated after 4 weeks.Results：The expression of human ILK gene was suppressed specific and effectively by transfection miRNA plasmid into cells.The inhibition efficiency of miR-3 is the strongest of 4 sequence.Tumor formation could be found in every group.The tumor in non-transfected group nude mice growth was faster than transfected group.In non-transfected group tumor tissue average volum was 289.56±36.49 mm3 and in transfected group was 56.67±4.32 mm3.Tumor tissue average weight of these two groups were 1.265±0.02g and 0.518±0.03g,and tumor growth was inhibited significantly（P0.05）.Conclusions：The expression of ILK in human bladder cancer cell lines BIU-87 can be inhibited specific and effectively by RNA interference that can result to tumor formation and growth suppressed obviously.

关 键 词：ILK BIU-87 裸鼠 

分 类 号：R735.35[医药卫生—肿瘤]