Shc3高表达抑制人肝癌细胞系凋亡并参与肝癌耐药  

Shc3 over-expression inhibitsapoptosis of HCC cells and is involved in drug resistance

作　　者：张新然 张弋[1] 高慧儿 强兆艳 李咏梅 ZHANG Xin-ran;ZHANG Yi;GAO Hui-er;QIANG Zhao-yan;LI Yong-mei(Department of Pharmacy, Tianjin First Central Hospital, Tianjin 300192;Department of Pharmacology, Basic Medicine College, Tianjin Medical University, Tianjin 300070;Department of Pathogen Biology, Basic Medicine College, Tianjin Medical University, Tianjin 300070, China)

机构地区：[1]天津市第一中心医院药学部,天津300192 [2]天津医科大学基础医学院药理学系,天津300070 [3]天津医科大学基础医学院病原生物学系,天津300070

出　　处：《基础医学与临床》2020年第4期473-478,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金青年科学基金(81800165)。

摘　　要：目的探讨Shc3对人肝细胞癌HCC细胞凋亡及耐药机制。方法选取稳定下调Shc3的人肝细胞癌系HCCLM3和HepG2细胞及其对照组scramble细胞,稳定上调Huh7和HepG2细胞及其对照组pCDH细胞。Western blot检测Shc3、MEK、p-MEK、ERK和p-ERK蛋白表达;real-time PCR检测Shc3 mRNA表达;凋亡实验检测细胞凋亡;CCK-8法检测细胞增殖。结果成功验证Shc3降表达及过表达细胞系;与scramble组比较,Shc3降表达组细胞MEK/ERK通路激活程度明显降低(P<0.05),细胞凋亡比例增加(P<0.05);与pCDH组比较,Shc3表达上调增加细胞对索拉菲尼耐受性(P<0.05),并且MEK/ERK通路激活程度明显增强(P<0.05)。结论Shc3可通过干扰HCC细胞凋亡,激活MEK/ERK通路,增加HCC细胞对索拉菲尼的耐药性,本研究结果为靶向Shc3治疗肝细胞癌提供了理论依据和实验室基础。Objective To investigate the apoptosis and drug resistance mechanism of Src homolog and collagen homolog 3(Shc3)in human hepatocellular carcinoma(HCC).Methods Human hepatocellular carcinoma cell lines HCCLM3 and HepG2 with stable Shc3 downregulation and their scramble cell control groups were selected,Huh7 and HepG2 cells with stable Shc3 up-regulation and their pCDH control groups were selected.Protein expression levels of Shc3,MEK,p-MEK,ERK and p-ERK were determined by Western blot analysis.The mRNA level of Shc3 were determined by real-time PCR.Apoptotic experiment was used to observe cell apoptosis.CCK-8 assays were used to observe cell proliferation.Results Shc3 down-regulation and up-regulation cell lines were proved.Compared with scramble cells,Shc3 down-regulation significantly decreased activity of MEK/ERK pathway(P<0.05),promoted cell apoptosis significantly(P<0.05).Compared with pCDH cells,Shc3 up-regulation increased celltolerance to sorafenib(P<0.05),Moreover,significantly enhanced activity of MEK/ERK pathway(P<0.05).ConclusionsShc3 may increase the resistance of HCC cells to sorafenib by interfering with apoptosis of HCC cells and activating MEK/ERK pathway.The results of this study provide theoretical basis and laboratory basis for targeting at Shc3 in the treatment of hepatocellular carcinoma.

关 键 词：人肝细胞癌 Shc3 细胞凋亡 细胞耐药 

分 类 号：R739.41[医药卫生—肿瘤]