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

Abstract

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. Key words: Vitamin K2; Phosphatidylcholine; Hepatocellular carcinoma cell; Bioinformatics; WNT3A; MicroRNA-16