siRNA干扰沉默KLK12基因抑制胃癌细胞生长的研究  

Experimental study on silencing the KLK12 expression by siRNA to inhibit the growth of gastric cancer cells

作　　者：沈志勇[1] 赵恩昊[1] 刘骅[1] 金鑫[1] 曹晖[1] 

机构地区：[1]上海交通大学医学院附属仁济医院普通外科,上海200127

出　　处：《中国普通外科杂志》2010年第10期1071-1075,共5页China Journal of General Surgery

基　　金：上海市科学技术委员会科研计划项目(08411964200)

摘　　要：目的利用RNA干扰(RNAi)技术,构建KLK12-siRNA表达载体,探讨其在体外对胃癌细胞生长的影响。方法合成KLK12-siRNA,以脂质体法转染至胃癌MKN-45细胞中建立稳定细胞系,并分成3组,即空白对照组、阴性对照组和KLK12siRNA组。采用实时定量PCR和Western blot方法观察KLK12siRNA转染后胃癌细胞KLK12基因和相应蛋白表达的变化;用MTT法,细胞迁移和侵袭试剂盒分别检测转染细胞的增殖、迁移和侵袭能力的变化。结果 KLK12siRNA组的KLK12mRNA和蛋白表达水平较两个对照组明显下调(P<0.05)。KLK12基因沉默显著抑制胃癌细胞的增殖,并降低体外胃癌细胞的迁移能力。结论 KLK12-siRNA表达载体能抑制胃癌MKN-45细胞的增殖和迁移,为胃癌的基因治疗提供了有益的实验依据。Objective To determine the inhibitory effect of synthetic KLK12 short interfering RNA on the expression of KLK12 gene in human gastric cancer line MKN-45 cells,and to elucidate its effect on proliferation of MKN-45 cells in vitro.Methods The constructed KLK12-siRNA plasmid was stably transfected into MKN-45 cells by lipofectamine,then,MKN-45 cells were divided into negative control group,blank control group and KLK12siRNA group.KLK12 expression was determined by real time PCR and Western blot.The proliferation,migration and invasion of transfected MKN-45 cells were evaluated by MTT assay,cell Migration and Invasion Assay Combo Kit.Results KLK12siRNA significantly inhibited the expression of KLK12 in gastric cancer cells at both mRNA and protein level.The proliferation of MKN-45 cells in KLK12siRNA group was significantly suppressed and the ability of migration was also significantly decreased in vitro.Conclusions The KLK12-siRNA expression vector can effectively suppress the proliferation and migration of MKN-45 cells,which may provide a novel gene therapy for gastric cancer.

关 键 词：胃癌 RNA干扰 细胞增殖 

分 类 号：R735.2[医药卫生—肿瘤]