eEF-2K基因沉默对肝癌细胞增殖、侵袭和凋亡的影响  

EFFECTS OF eEF-2K GENE SILENCING ON PROLIFERATION,INVASION AND APOPTOSIS OF HEPATOCELLULAR CARCINOMA CELLS

作　　者：肖瑞雪 路帅 杨鹏辉 康毅敏[1] XIAO Ruixue;LU Shuai;YANG Penghui;KANG Yimin(Key Laboratory of Molecular Pathology,Inner Mongolia Medical University,Hohhot 010059,China;Beijing Shijitan Hospital of Capital Medical University,Beijing 100038,China;The First Medical Center,PLA General Hospital,Beijing 100853,China)

机构地区：[1]内蒙古医科大学分子病理学重点实验室,内蒙古呼和浩特010059 [2]首都医科大学附属北京世纪坛医院,北京100038 [3]解放军总医院第一医学中心,北京100853

出　　处：《内蒙古医科大学学报》2023年第3期240-245,共6页Journal of Inner Mongolia Medical University

基　　金：内蒙古医科大学致远人才计划治学人才团队项目(ZY0130011)。

摘　　要：目的检测eEF-2K基因在肝细胞癌(HCC)组织内的表达并分析其表达水平对HCC细胞增殖、侵袭和凋亡的影响。方法用RT-qPCR和Western Blot检测了64例肝癌组织及癌旁肝组织,5种HCC细胞系LM3、Hep3B、Huh7、97H、HepG2及人正常肝细胞系LO2中eEF-2K基因在mRNA及蛋白水平的表达。用eEF-2K特异性siRNA沉默LM3及Hep3B细胞性中eEF-2K基因后,经qPCR检测确认。随后用CCK-8、Transwell方法及流式细胞技术检测eEF-2K基因沉默对增殖、侵袭与凋亡的影响。结果相较癌旁组织,HCC组织内eEF-2K mRNA与蛋白表达显著上调,差异有统计学意义(P<0.05);与正常人肝细胞系LO2相比,肝癌细胞系LM3、Hep3B、97H、HepG2、Huh7中的eEF-2K mRNA和蛋白表达分别升高,差异有统计学意义(P<0.05)。在LM3及Hep3B细胞系中用特异性siRNA沉默eEF-2K后,与非特异性siRNA对照组相比,细胞增殖活性和侵袭能力显著降低(P<0.05),凋亡率显著升高,差异有统计学意义(P<0.05)。TCGA数据库分析结果表明eEF-2K高表达与肝癌预后不良相关,差异有统计学意义(P<0.05)。结论eEF-2K蛋白在肝癌组织和细胞中高表达且与预后不良相关;沉默eEF-2K后肝癌细胞增殖和侵袭能力降低并诱发凋亡,提示eEF-2K可用作HCC潜在的预后标志物和治疗靶点。Objective To detect the expression level of eEF-2K in hepatocellular carcinoma(HCC)and the effect of its silencing on proliferation,invasion and apoptosis of HCC cells.Methods RT-qPCR and Western Blot were used to examine the expression of eEF-2K gene at mRNA and protein levels in HCC and their adjacent liver tissues resected from 64 HCC pa-tients,as well as in five HCC lines(LM3,Hep3B,Huh7,97H,HepG2)and normal human liver cell line LO2.LM3 and Hep3b cells were transfected with eEF-2K specific and non-specific siRNAs respectively.The silencing of eEF-2K gene after siRNA trasfection in LM3 and Hep3B cell lines was confirmed by RT-qPCR.The effect of silencing of eEF-2K gene on proliferation,invasion and apoptosis of HCC were detected by CCK-8 assay,Transwell assay and flow-cytometry.Results The expression of eEF-2K was significantly up-regulated both at mRNA and protein levels in HCC tissues as well as in LM3,HEP3B,97H,Hepg2,Huh7 lines compared to the expressions in adjacent tissues(P<0.05)and in normal LO2 line(P<0.05).The prolifera-tion rates of eEF-2K-silenced groups were decreased compared to control group(P<0.05),and the percentages of transmem-brane cells in eEF-2K-silenced groups groups were decreased compared to control group(P<0.05).Silencing of eEF-2K by siRNA also led to increased apoptosis in HCC(P<0.05).The analysis of TCGA database showed that the high expression of eEF-2K was associated with poor prognosis of HCC(P<0.05).Conclusions eEF-2K was highly expressed in HCC tissues and cell lines,which is associated with poor prognosis.Knock-down of eEF-2K gene by siRNA inhibited the proliferation and invasion ability,and induced apoptosis in HCC cells.This study provided evidence that eEF-2K is potential prognostic marker and therapeutic target for HCC.

关 键 词：肝细胞癌 eEF-2K SIRNA 增殖 侵袭 凋亡 

分 类 号：R363.1[医药卫生—病理学]