沉默MCM7基因后肝癌SMMC-7721细胞差异表达基因的筛选  

Screening of differentially expressed genes after silencing MCM7 in liver cancer cell line SMMC-7721

作　　者：罗旺[1,2] 曹骥[1] 杨香娣 邓玲[1,2] 王谷洋 杨春[1] 李科志[1] 李瑗[1] 

机构地区：[1]广西医科大学附属肿瘤医院实验研究部,广西壮族自治区南宁市530021 [2]广西医科大学研究生学院,广西壮族自治区南宁市530021

出　　处：《世界华人消化杂志》2016年第10期1492-1500,共9页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.81260080;广西自然科学基金资助项目;No.2013GXNSFAA019210~~

摘　　要：目的:探究微小染色体维持蛋白7(minichromosome maintenance protein 7,MCM7)基因在调节肝癌细胞生长的相关机制.方法:通过s i R N A干扰技术沉默人肝癌SMMC-7721细胞中的MCM7基因表达,采用人全基因组表达谱芯片,筛查MCM7沉默后差异表达基因,进行生物信息学分析,并对部分差异表达基因进行蛋白免疫印迹法(Western blot)验证.结果:芯片筛选出在人肝癌SMMC-7721细胞中沉默MCM7基因后差异表达基因1010个,其中上调基因391个,下调基因619个.这些基因主要涉及生物大分子代谢、细胞周期调控、细胞增殖、细胞凋亡等方面,参与胞吞、肿瘤相关通路、P53信号通路、mTOR信号通路等通路的改变.Western blot对部分差异基因表达验证的结果显示CCND1、SKP2和JUP蛋白表达均下调,与芯片检测结果一致.结论:应用人基因表达谱芯片,成功筛选出沉默MCM7基因后人肝癌细胞SMMC-7721差异表达基因,为探究MCM7基因影响肝癌细胞生长提供了有效线索.AIM： To explore the mechanisms of minichromosome maintenance protein 7（MCM7） for regulating the growth of liver cancer cells.METHODS： The expression of MCM7 gene in SMMC-7721 cells was silenced with small interfering RNA（si RNA）. Human genomewide expression profilechip was then employed to screen the differentially expressed genes, and bioinformatics analysis of the differentially expressed genes was performed. Finally, part of these differentially expressed genes were confirmed by Western blot assay. RESULTS： In total there were 1010 genes that were differentially expressed in SMMC-7721 cells after the expression of MCM7 was silenced, including 391 up-regulated and 619 downregulated ones. Bioinformatics analysis showed that these differentially expressed genes are involved in many cellular biological processes such as macromolecular metabolism, cell cycle regulation, cell proliferation regulation,apoptosis, endocytosis, P53 and mTOR signaling pathways. The down-regulation of CCND1, SKP2 and JUP was confirmed by Western blot, which was consistent with the results of the genome-wide expression profile chip.CONCLUSION： The differentially expressed genes after silencing the gene MCM7 in liver cancer cells SMMC-7721 might provide some clues for understanding the mechanism by which MCM7 affects the growth of liver cancer cells.

关 键 词：肝癌 MCM7 RNA干扰 人基因表达谱芯片 

分 类 号：R735.7[医药卫生—肿瘤]