siRNA靶向抑制GHR对人结肠癌SW480细胞增殖的影响  

Effect of siRNA targeting inhibition of growth hormone receptor on proliferation of human colon cancer cell line SW480

作　　者：周东[1,2] 张毅[2] 梁道明[2] 袁勇[1,2] 李思齐[2] 曾德妙[1] 陈嘉勇[2] 

机构地区：[1]昆明医学院第二临床学院,云南昆明650101 [2]昆明医学院第二附属医院急诊科,云南昆明650101

出　　处：《中国普通外科杂志》2011年第9期945-950,共6页China Journal of General Surgery

基　　金：云南省应用基础面上项目(2009ZC106M)

摘　　要：目的利用小分子干扰RNA（siRNA）技术抑制人结肠癌SW480细胞人生长激素受体（GHR）基因的表达,观察GHR基因沉默对SW480细胞增殖的影响。方法设计特异性siRNA,构建针对人GHR真核质粒的表达载体（pcDNATM6.2-GW/EmGFP-siRNA-GHR）,用电转染法转染人结肠癌SW480细胞,培养48~72 h后,用四甲基偶氮唑蓝（MTT）法检测细胞增殖活性;Western Blot检测GHR蛋白表达;实时荧光定量PCR（qPCR）方法检测GHRmRNA水平变化。结果转染siRNA后,与对照组相比,转染组细胞增殖速率、GHR的蛋白表达量、GHRmRNA水平均明显降低（均P〈0.05）。结论构建的pcDNATM6.2-GW/EmGFP-GHRsiRNA能有效地下调SW480细胞中GHR的表达,从而抑制SW480细胞的增殖。Objective To inhibit growth hormone receptor（GHR） gene expression in human colon cell line SW480 by small interfering RNA（siRNA） technique,and thereby to observe the effect of GHR gene silencing on proliferation of SW480 cells.Methods After the GHR gene specific siRNA design,the eukaryotic plasmid expression vector targeting human GHR（pcDNATM6.2-GW/EmGFP-siRNA-GHR） was constructed,which was then transfected into SW480 cells by electroporation.The transfected cells were cultured for 48 to 72 hours,and then the cell proliferative activity was assayed by tetrazolium bromide（MTT）,GHR protein expression was detected by Western Blot,and GHR mRNA was determined by fluorescence real time quantitative reverse transcription-polymerase chain reaction,respectively.Results Compared with the control group,the proliferative speed,GHR protein and mRNA expression of SW480 cells were all significantly decreased after transfection（all P0.05）.Conclusions The pcDNATM6.2-GW/EmGFP-GHR-siRNA constructed can effectively down-regulate GHR expression in SW480 cells,and thereby inhibit the proliferation of SW480 cells.

关 键 词：结肠肿瘤 RNA 小分子干扰 受体 人生长激素 

分 类 号：R735.3[医药卫生—肿瘤]