组织蛋白酶S表达降低致肝癌细胞凋亡的机制研究  

Detection of apoptosis mechanisms in hepatocellular carcinoma regulated by the knockdown of cathepsin S

作　　者：董小锋[1] 王付海[2] 李涛[2] 钟敬涛[2] 修鹏[2] 刘锋[2] 徐宗珍[2] 李杰[2] 

机构地区：[1]广西壮族自治区人民医院肝胆外科,广西南宁530021 [2]山东大学附属千佛山医院肝胆外科,山东济南250014

出　　处：《中国现代普通外科进展》2014年第11期841-845,共5页Chinese Journal of Current Advances in General Surgery

基　　金：山东省医药卫生科技发展计划(2009HZ070);山东省科技攻关项目(2010GSF10230)

摘　　要：目的:探讨组织蛋白酶S(Cathepsin S,Cat S)表达降低后对肝癌细胞凋亡的影响及其机制。方法:选择2种肝癌细胞系MHCC97-H、Bel-7402,将肝癌细胞分为对照组、Con sh RNA组、Cat S sh RNA组。慢病毒介导的Cat S sh RNA转染肝癌细胞48 h后,Western blot检测Cat S表达变化;Annexin V/PI双染流式测细胞凋亡;Western blot检测内源性凋亡相关蛋白Bax、Bcl-2表达变化;Western blot检测外源性凋亡相关蛋白Fas、Fas-L表达变化。结果:在肝癌细胞MHCC97-H、Bel-7402中,Cat S sh RNA可显著降低肝癌细胞中Cat S表达(P<0.01);Cat S表达降低后,凋亡细胞显著增多(P<0.01);内源性促凋亡蛋白Bax表达升高(P<0.01),抗凋亡蛋白Bcl-2表达降低(P<0.01);外源性促凋亡蛋白Fas-L表达明显增强(P<0.01),Fas表达无明显变化(P>0.05)。结论:Cat S可同时通过内源性及外源性凋亡途径对肝癌细胞的凋亡进行调控。Objective： To detect the apoptosis effects and possible mechanisms mediated by the knockdown of cathepsin S in hepatocellular carcinoma cell line. Methods： HCC cell line MHCC97-H, Bel-7402 were selected and divided into three group： Control group; Con shRNA group; Cat S shRNA group. Lentivirus mediated Cat S shRNA was transfected into HCC cells, 48 h later, the cells were collected. The expression of Cat S, intrinsic apoptosis pathway associated pro- tein Bax, Bcl-2 and extrinsic apoptosis pathway associated protein Fas-L, Fas was detected by Western blot. The cell apoptosis rate was detected by annexin V/PI double staining via flow cy- tometer. Results： Compared with the control group and con shRNA group,Cat S shRNA signifi- cantly decreased Cat S expression in both HCC cell lines （P〈0.01）; there were more apoptosis cells in Cat S shRNA group compared with the other two groups （P〈0.01）;the expression of intrinsic pro-apoptotic protein Bax and extrinsic pro-apoptotic Fas-L was significantly up-regulated in Cat S shRNA group （P〈0.01）; The expression of intrinsic anti-apoptotic protein Bcl-2 was significantly down-regulated in Cat S shRNA group（P〈0.01）; there was no significant difference on the expres- sion of extrinsic pro-apoptotic protein Fas-L in shRNA group and the other two groups（ P〉0.05）. Conelusion： The knockdown of Cat S could regulate HCC cell apoptosis through both intrinsic and extrinsic apoptosis signal dependent pathways.

关 键 词：肝细胞肝癌 组织蛋白酶S 内源性凋亡通路 外源性凋亡通路 SHRNA 

分 类 号：R735.7[医药卫生—肿瘤]