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

发布时间：2018-03-25 11:52

本文选题：肝细胞癌　切入点：SIRT　出处：《中国细胞生物学学报》2016年07期

【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-sh RNA-SK-Hep-1和sh Cont-SKHep-1,并应用定量逆转录PCR(q RT-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q RT-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-sh RNASK-Hep-1组裸鼠移植瘤体积和质量较sh Cont-SK-Hep-1组均减少(P0.01);免疫组织化学发现,SIRT6-sh RNA-SK-Hep-1组Ki-67水平下调;沉默SIRT6基因下调XIAP m RNA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。
[Abstract]:The aim of this study was to investigate the effect of silencing of 6(silent information regulator 6 sir6) gene silencing on proliferation and apoptosis of heterotopic transplanted human hepatoma cells in nude mice. Stable cell lines SIRT6-sh RNA-SK-Hep-1 and sh Cont-SKHep-1 were constructed and detected by quantitative reverse transcription PCR(q RT-PCR and Western blot. The expression level of SIRT6 gene; The stable transfected cells were injected subcutaneously into nude mice, and the growth of xenografts in nude mice was monitored in real time. After 7 weeks, the xenografts were stripped out and weighed. The protein level of SIRT6KI-67 was analyzed by immunohistochemistry. The effect of SIRT6 gene silencing on the downstream inhibitor of apoptosis proteinsi IAPs was further detected by Q RT-PCR. Western blot was used to detect X-linked inhibitor of apoptosis protein. Protein levels of gene XIP and poly ADP-ribose polymerase of polyadenosine diphosphate. SK-Hep-1 stable cell line with silencing SIRT6 gene was successfully constructed. The volume and mass of xenografts in the RNASK-Hep-1 group were lower than those in the sh Cont-SK-Hep-1 group, and the Ki-67 level was down-regulated in the SIRT6-sh RNA-SK-Hep-1 group, and the XIAP m RNA and protein levels were down-regulated by silencing SIRT6 gene. The results suggested that the silencing of SIRT6 gene might inhibit the growth of xenotransplantation of human hepatoma cells by down-regulating the expression of XIAP in nude mice.
【作者单位】：重庆医科大学感染性疾病分子生物学教育部重点实验室;
【基金】：国家自然科学基金(批准号:81472271、81270559)资助的课题~~
【分类号】：R735.7

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