Effect of miR-4698 mediated GALNT4 expression on proliferation and migration of hepatocellular carcinoma cells / 中国医师杂志

ABSTRACT

Objective:The relative expression of miR-4698 in liver cancer tissues and cell lines was detected, and its effect on the proliferation and migration of liver cancer cells and its molecular mechanism were analyzed.Methods:Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to analyze the relative expression of miR-4698 in liver cancer tissues and liver cancer cell lines. Among the lowest-expressing hepatocellular carcinoma cell lines, miR-4698 mimic and control mimic were transfected with liposome transfection method, and named as experimental group and control group. qRT-PCR was used to detect transfection efficiency. Cell counting kit-8 (CCK-8) and transwell migration experiments were used to detect the effects of overexpressing miR-4698 on the proliferation and migration ability of liver cancer cells. Bioinformatics and dual luciferase reporter gene experiments were used to predict and verify the binding of miR-4698 to target gene. qRT-PCR and Western blot were used to detect the relative expression of target gene at mRNA and protein levels, respectively.Results:Compared with the adjacent tissues, miR-4698 was significantly lower in the liver cancer ( P<0.01). Compared with normal hepatocytes, miR-4698 was significantly lower in hepatoma cell lines ( P<0.05), and the lowest in Huh7 cells ( P<0.01). After transfection, the expression of miR-4698 in Huh7 cells in experimental group was significantly increased compared with that in the control group ( P<0.01). Overexpression of the miR-4698 can inhibit the proliferation and migration of Huh7 cells ( P<0.05). Bioinformatics showed that the target gene of miR-4698 was polypeptide-n-acetylgalactosamine transferase 4 (GALNT4), and the double luciferase reporter gene confirmed that miR-4698 could bind to GALNT4 ( P<0.01). qRT-PCR showed that overexpression of miR-4698 could inhibit the expression of GALNT4 gene ( P<0.01). Western blot results showed that overexpression of miR-4698 decreased the protein expression of GALNT4, cyclin B and CDK1, and increased the protein expression of N-cadherin and ZEB-2. Conclusions:The expression of miR-4698 in liver cancer tissues and cell lines is significantly reduced. Overexpression of miR-4698 can inhibit the expression of GALNT4 gene and reduce the proliferation and migration ability of liver cancer Huh7 cells.