SIRT6基因沉默对人肝癌细胞凋亡的影响及其机制研究  

Effects of SIRT6 Gene Silencing on the Apoptosis in Human Hepatocellular Carcinoma Cells and Its Potential Mechanisms

作　　者：周洪钟[1] 陶娜娜[1] 陈祥[1] 任吉华[1] 李宛蔚[1] 刘波[1] 陈娟[1] 

机构地区：[1]重庆医科大学感染性疾病分子生物学教育部重点实验室,重庆400016

出　　处：《中国细胞生物学学报》2016年第4期389-396,共8页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81472271;81270559)资助的课题~~

摘　　要：该文旨在探讨慢病毒介导的沉默信息调节因子6(silent information regulator 6,SIRT6)基因沉默对人肝癌细胞凋亡的影响及其机制。逆转录PCR(RT-PCR)和Western blot分别检测人肝癌细胞系(SK-Hep-1、Huh-7、PLC/PRF/5、Hep G2)和永生化肝细胞系(MIHA)中SIRT6基因的表达水平;利用慢病毒介导的sh RNA干扰技术靶向沉默SIRT6的表达,并通过RT-PCR和Western blot验证其沉默效率;流式细胞术检测SIRT6基因沉默对人肝癌细胞凋亡的影响,进一步应用RT-PCR和Western blot检测SIRT6基因沉默对凋亡抑制蛋白基因(inhibitor of apoptosis proteins,IAPs)家族m RNA和蛋白质水平的影响;最后,应用流式细胞术分析X连锁凋亡抑制蛋白基因(X-linked inhibitor of apoptosis protein gene,XIAP)在SIRT6基因沉默诱导的肝癌细胞凋亡中的作用。结果显示,SIRT6基因在人肝癌细胞系中表达上调;慢病毒介导的sh RNA能抑制人肝癌细胞中SIRT6基因的表达;沉默SIRT6基因的表达能诱导人肝癌细胞凋亡,并降低XIAP的m RNA和蛋白质水平;过表达XIAP能逆转SIRT6基因沉默所诱导的人肝癌细胞凋亡。该研究结果提示,SIRT6基因沉默可能通过调节XIAP的表达从而诱导人肝癌细胞凋亡。This study investigated the effects of lentivirus-mediated silent information regulator 6（SIRT6） knock-down on cell apoptosis in hepatocellular carcinoma（HCC） cells and its potential mechanisms. The expression of SIRT6 gene in HCC cell lines（SK-Hep-1, Huh-7, PLC/PRF/5, Hep G2） and immortalized human liver cell line（MIHA） was detected by reverse-transcription polymerase chain reaction（RT-PCR） and Western blot, respectively. SIRT6 gene was silenced by lentivirus-mediated sh RNA interference technology and its efficiency of SIRT6 gene silencing was detected by RT-PCR and Western blot. Effects of SIRT6 gene silencing on apoptosis in SK-Hep-1 and Huh-7 cells were analyzed by flow cytometry. The m RNA and protein levels of IAPs（inhibitor of apoptosis proteins） family were analyzed by RT-PCR and Western blot, respectively. Effects of XIAP（X-linked inhibitor of apoptosis protein gene） on the induction of apoptosis in SIRT6-depleted SK-Hep-1 cells were analyzed by flow cytometry. Our results showed that SIRT6 was over-expressed in HCC cells. Lentivirus-mediated sh RNA interference down-regulated the protein level of SIRT6 in the HCC cells. Knockdown of SIRT6 gene induced the apoptosis of HCC cells and down-regulated the m RNA and protein levels of XIAP. XIAP over-expression significantly abolished the induction of apoptosis in SIRT6-depleted cells. Together, these data indicated that SIRT6 gene silencing might induce cell apoptosis in human hepatocellular carcinoma cells by regulating XIAP protein level.

关 键 词：肝细胞癌 SIRT6 XIAP 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]