Mechanism of Apaf-1 for regulating liver cancer by wnt/β-catenin signaling pathway / 重庆医学

ABSTRACT

Objective To study the rule of Apaf-1 in wnt/β-catenin signaling pathway and its regulatory expression mechanism in hepatocellular carcinoma cells.Methods The regulatory mechanism of Apaf-1 gene in Wnt/β-catenin signaling pathway was verified by the TOPflash experiment;real-time PCR was used to detect the expression amounts of Apaf-1 in various hepatoma cell lines (HepG2,H HCC,HB611) and normal liver cell line(LO2);the RNAi interference plasmid of Apaf-1 was constructed and transfected into the HepG2 cell line.Then the transfection efficiency and expression of related genes and proteins were detected.Results The TOPFlash experiment found that Apaf-1 gene could inhibit the wnt/β-catenin signal pathway in a dose dependent manner;the Apaf-1 expression level in hepatoma cells was decreased compared with the normal liver cells,moreover its expression level was lowest in HepG2 cell line;RNA interfering Apaf-1 gene in HepG2 cell line,the expression level of Apaf-1 gene was significantly decreased,and the expression levels of downstream genes and protein(β-catenin,Cyclin A,CDK2,wnt5a,STAT3,EGFR,APC) in wnt signal pathway were significantly increased or decreased,the difference was statistically significant(P＜0.05).Conclusion Apaf-1 gene plays an important role in the formation process of hepatocarcinoma cells by wnt/β-catenin signaling pathway.