蛋白激酶B(PKB/AKT1)小干扰RNA通过WNT/β-Catenin通路抑制人胃癌细胞株BGC-823的增殖与迁移  

siRNA of PKB/AKT1 inhibits proliferation and migration of BGC-823 cells via WNT/β-catenin signaling pathway

作　　者：李树静 张志成 林佳 高超 王忠利 Shu-jing Li Zhi-cheng Zhang Jia Lin Chao Gao Zhong-li Wang(Department of General Surgery, the First Affiliated Hospital of Jinzhou Medical University, Jinzhou, Liaoning 121001, China)

机构地区：[1]锦州医科大学附属一院外科,辽宁锦州121001

出　　处：《中国现代医学杂志》2017年第2期31-35,共5页China Journal of Modern Medicine

基　　金：2014年省级大学生创新创业训练计划项目(No:201410160032)

摘　　要：目的观察蛋白激酶B(PKB/AKT1)小干扰RNA(si RNA)对人胃癌细胞株BGC-823增殖与迁移的影响,探讨PKB/AKT1在胃癌发生中的可能作用机制。方法体外培养BGC-823细胞,分为空白对照组、阴性对照组和PKB/AKT1 si RNA转染组。实时荧光定量聚合酶链反应(q RT-PCR)和Western blot法分别检测BGC-823细胞PKB/AKT1的m RNA和蛋白表达;噻唑蓝法测定细胞增殖抑制率;划痕实验测定细胞迁移。Western blot法检测β-Catenin,侵袭转移相关的上皮性钙黏附蛋白(E-Cadherin)、神经性钙黏附蛋白(N-Cadherin)和基质金属蛋白酶(MMP-9)的表达。结果与两组对照组比较,PKB/AKT1 si RNA转染组PKB/AKT1的m RNA和蛋白表达降低,并抑制BGC-823细胞的增殖和迁移,β-Catenin,N-Cadherin及MMP-9的蛋白表达下降,E-Cadherin表达增加。结论 PKB/AKT1 si RNA抑制胃癌细胞株BGC-823的增殖和迁移,其机制可能与si RNA下调PKB/AKT1表达进而抑制WNT/β-Catenin信号通路有关。Objective To observe the effect of double-stranded small interfering RNA （siRNA） of protein kinase B （PKB/AKT1） on gastric cancer cell line BGC-823 cell proliferation and migration, and to explore the possible underlying mechanism. Methods BGC-823 cells were cultured in vitro and then randomly divided into blank group, negative control group and AKT1 siRNA-transfected group. AKT1 siRNA efficiency was examined through qRT-PCR and Western blot analysis. The proliferation rate of BGC-823 cells was determined through a methyl thiazolyl tetrazolium assay, and the cell migration of BGC-823 cells was examined by scratch assay. The β-catenin, E- Cadherin, N-Cadherin and MMP-9 protein expression levels were determined using Western blot. Results Compared with those of the blank and negative control groups, the expression levels of AKT1 mRNA and protein significantly decreased in the AKT1 siRNA-transfected cells. In addition, the proliferation rate and migration of BGC-823 cells were markedly decreased. The expression level of E-Cadherin increased, whereas β-catenin, N-Cadherin and MMP-9 protein levels were inhibited in the AKT1 siRNA-transfeeted BGC-823 ceils. Conclusions siRNA of AKT1 inhibits BGC-823 cell proliferation and migration via WNT/β-catenin signaling pathway.

关 键 词：蛋白激酶B 胃癌 BGC-823 WNT／β-Catenin 

分 类 号：R581.3[医药卫生—内分泌]