siRNA干扰Gab1表达对胆管癌细胞株HUCCA-1功能的影响及其与PI3K/Akt信号通路的关系  

Effect of silencing Gab1 expression on cholangiocarcinoma cell line HUCCA-1 and its correlation with PI3K/Akt pathway

作　　者：姚旭[1] 田忠[1] 刘源[1] 

机构地区：[1]中国医科大学附属盛京医院第十普外科,沈阳110042

出　　处：《天津医药》2015年第6期590-594,I0003,共6页Tianjin Medical Journal

基　　金：辽宁省科技攻关计划项目(2013408001)

摘　　要：目的：检测下调Gab1表达后人胆管癌细胞株HUCCA-1细胞功能、PI3KCA及Akt1的蛋白及mRNA表达水平改变，探讨Gab1、PI3KCA及Akt1表达在胆管癌恶性行为中的作用机制。方法 Gab1siRNA转染HUCCA-1细胞，采用qRT-PCR法及Western blot法检测转染效率及PI3KCA、Akt1表达水平，MTT检测转染后细胞增殖变化，流式细胞术检测转染后细胞凋亡变化，Transwell检测细胞迁移及侵袭能力。结果 Gab1siRNA在HUCCA-1细胞中的转染效率约为65%～70%，转染效率佳；Gab1siRNA转染HUCCA-1细胞后，PI3KCA、Akt1的蛋白及mRNA表达量下调；Gab1siRNA组HUCCA-1细胞增殖能力在48 h、72 h及96 h均低于siRNA control组和Mock组（P〈0.05）；Gab1siRNA组的HUCCA-1细胞凋亡率高于Mock及siRNA control组（P〈0.05）；Gab1siRNA组HUCCA-1细胞迁移减少百分比及侵袭能力低于siRNA control组及Mock组（P＜0.05）。结论 Gab1可能通过激活PI3K/Akt信号通路表达促进肿瘤细胞恶性行为，Gab1可作为胆管癌治疗新的靶向标志物及候选基因。Objective To explore the functionalternation of human cholangiocarcinoma cell line HUCCA-1 by silenc-ing Gab1 expression;to detect its effect on PI3KCA and Akt1 expression at protein and mRNA levels and to explore the role of Gab1,PI3KCA and Akt1 expressions in the malignant behavior of cholangiocarcinoma. Methods Gab1 siRNA was trans- fected into cholangiocarcinoma cell line HUCCA-1. Proliferation, apoptosis, migration/invasion of cells were examined by MTT, flow cytometry and Transwell assay respectively after transfection. PI3KCA and Akt1 expressions were detected by Western blotting and qPCR. Results Transfection efficiency was satisfactory and reaches 65%~70%;PI3KCA and Akt1 expressions were down-regulated at both protein and mRNA levels upon silencing of Gab1. Meanwhile, proliferations of HUCCA-1 cells were inhibited （P〈0.01）;apoptosis rate increases（P〈0.05）;and migration and invasion were both inhibit-ed upon Gab1 silencing（P〈0.05）. Conclusion The proliferation, migration and invasion are all inhibited while apoptosis is attenuated after Gab1 was down-regulated by Gab1 siRNA transfection. PI3KCA and Akt1 expressions are up-regulated with down-regulation of Gab1. Therefore, Gab1 may enhance the maglinant behavior of cholangiocarcinoma cells through up-regulating PI3KCA and Akt1 expression and it can be used as a candidate for cholangiocarcinoma therapy.

关 键 词：胆管癌 增殖 凋亡 Gab1 HUCCA-1 PI3K/AKT信号通路 

分 类 号：R735.8[医药卫生—肿瘤]