维生素K2联合磷脂酰胆碱抑制人肝癌细胞作用机制的生物信息学分析  

Bioinformatics analysis of inhibitory mechanism of vitamin K2 combined with phosphatidylcholine on human hepatocellular carcinoma cells

作　　者：闫瑞承[1] 罗建飞[1] 沈世强[2] Yan Ruicheng;Luo Jianfei;Shen Shiqiang(Department of Gastrointestinal Surgery East Section,Renmin Hospital of Wuhan University,Wuhan,430205,China;Department of Hepatobilitary Surgery,Renmin Hospital of Wuhan University,Wuhan,430060,China)

机构地区：[1]武汉大学人民医院东院胃肠外科,430205 [2]武汉大学人民医院肝胆外科,430060

出　　处：《中华实验外科杂志》2019年第10期1738-1740,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨维生素K2联合磷脂酰胆碱抑制人肝癌细胞的分子机制.方法维生素K2联合磷脂酰胆碱处理人肝癌细胞(SMMC-7721)48 h,使用微小RNA(miRNA,miR)芯片检测处理后的人肝癌细胞miRNA表达水平,采用生物信息学预测miRNA的靶基因及靶基因所参与的功能分类、细胞信号通路,最后通过免疫蛋白印迹实验验证细胞信号通路的关键蛋白表达.结果共有65个miRNA表达水平被显著上调或下调2倍以上,miR-16的表达水平变异最大,对比对照组上调了8 171.07倍(t=6.238,P<0.01).65个miRNA分别有8~326个靶向基因,主要分布于上调部分和生物过程,其中miR-16有164个靶向基因.将靶向基因映射到信号通路后,Wnt信号通路富集靶基因最显著,有58个靶向基因(t=8.753,P<0.01).免疫蛋白印迹实验中WNT3A和磷酸化β-连环蛋白(β-catenin)的低表达证实了经典Wnt信号通路参与了维生素K2联合磷脂酰胆碱抑制人肝癌细胞的过程(t=5.354,P<0.01).结论维生素K2联合磷脂酰胆碱通过上调miR-16表达,抑制Wnt信号通路,达到抑制人肝癌细胞的效果.Objective To explore the molecular mechanism of vitamin K2 (vk2) combined with phosphatidylcholine (pc) inhibiting human hepatocellular carcinoma cells. Methods Vitamin K2 combined with phosphatidylcholine was used to treat human hepatocellular carcinoma cells (SMMC-7721) for 48 hours. The expression of microRNA (miRNA, miR) in the treated human hepatocellular carcinoma cells was detected by microarray. Bioinformatics was used to predict the functional classification and cell signaling pathway and target genes of miRNA. Finally, the key proteins of cell signaling pathway were verified by Western blotting. Results A total of 65 miRNAs were significantly up-regulated or down-regulated by more than two folds. The variation of the expression level of miRNA16 was the greatest, up-regulated by 8 171.07 folds compared with the control group (t=6.238, P<0.01). There were 8 to 326 targeting genes in 65 miRNAs, which were mainly distributed in up-regulation and biological processes. Among them, there were 164 targeting genes in miR-16. After mapping the target genes to the signaling pathways, Wnt signaling pathway enriched the target genes most significantly, with 58 target genes (t=8.753, P<0.01). The low expression of WNT3A and phosphorylated β-catenin in Western blotting confirmed that the classical Wnt signaling pathway was involved significantly in the human hepatocellular carcinoma cells inhibited by vitamin K2 combined with phosphatidylcholine (t=5.354, P<0.01). Conclusion Vitamin K2 combined with phosphatidylcholine can inhibit the Wnt signaling pathway by up-regulating the expression of miR-16 in human hepatocellular carcinoma cells.

关 键 词：维生素K2 磷脂酰胆碱 肝癌细胞 生物信息学 WNT3A 微小RNA-16 

分 类 号：R735.7[医药卫生—肿瘤]