sirt6基因沉默对裸鼠异位移植人肝癌细胞增殖和凋亡的影响.pdfVIP

中国细胞生物学学报 Chinese Journal of Cell Biology 2016, 38(7): 82 1–828 DOI: 10.11844/cj cb.2016.07.0066 SIRT6基因沉默对裸鼠异位移植人肝癌细胞 增殖和凋亡的影响 周洪钟 陶娜娜 陈 祥 任吉华 李宛蔚 刘 波 陈 娟* (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) 摘要 该文旨在探讨沉默信息调节因子6(silent information regulator 6, SIRT6)基因沉默对裸 鼠异位移植人肝癌细胞增殖和凋亡的影响。构建稳定细胞系SIRT6-shRNA-SK-Hep-1和shCont-SK- Hep-1, 并应用定量逆转录PCR(qRT-PCR)和Western blot检测SIRT6基因的表达水平; 将稳定转染细 胞注入裸鼠皮下, 实时监测裸鼠移植瘤的生长, 7周后剥离出裸鼠移植瘤并称重; 免疫组织化学分析 SIRT6、Ki-67的蛋白质水平。进一步应用qRT-PCR检测SIRT6基因沉默对下游靶向分子凋亡抑制 蛋白(inhibitor of apoptosis proteins, IAPs) 家族的影响; Western blot检测X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein gene, XIAP)和多腺苷二磷酸核糖聚合酶(poly ADP-ribose polymerase, PARP)蛋白质水平。结果显示, 成功构建了沉默SIRT6基因的SK-Hep-1稳定细胞系; SIRT6-shRNA- SK-Hep-1组裸鼠移植瘤体积和质量较shCont-SK-Hep-1组均减少(P0.01); 免疫组织化学发现, SIRT6-shRNA-SK-Hep-1组Ki-67水平下调; 沉默SIRT6基因下调XIAP mRNA和蛋白质水平, 并增加 PARP蛋白质的剪切水平。该研究结果提示, SIRT6基因沉默可能通过下调XIAP的表达抑制裸鼠异 位移植人肝癌细胞的生长。 关键词 肝细胞癌; SIRT6; XIAP; 移植瘤; 生长 Effects of SIRT6 Gene Silencing on Human Hepatocellular Carcinoma Cell Proliferation and Apoptosis in Ectopic Xenograft Nude Mouse Model Zhou Hongzhong, Tao Nana, Chen Xiang, Ren Jihua, Li Wanyu, Liu Bo, Chen Juan* (Key Laboratory of Molecular Biology on Infection Diseases of Ministry of Education, Chongqing Medical University, Chongqing 400016, China) Abstract This study investigated the effects of SIRT6 gene silencing on human hepatocellular carcinoma cell proliferation and apoptosis in ectopic xenograft nude mouse model. SIRT6-shRNA-SK-Hep-1 and shCont-SK- Hep-1 stable cell lines were established. The mRNA and protein levels of SIRT6 gene were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR) and Western blot, respectively. The stab