TIMP-3 regulated by miR-181b as a target gene on invasion and migration of hepatocellular carcinoma cells / 中华肝胆外科杂志

ABSTRACT

Objective To explore the impact of TIMP 3 regulated by miR-181b as a target gene on invasion and migration of hepatocellular carcinoma (HCC) in vitro.Methods The expressions of miR-181b were detected using SYBR Green real-time fluorescence quantitative polymerase chain reaction on liver cancer specimens and on HCC cell lines.The protein expression of TIMP 3 in HCC was detected using westen blot,and SKHep-1 as a cell line expressing high miR-181b was chosen through reporter gene experiment.TIMP-3 as a target gene regulated by miR-181b and its effect on invasion and migration treated by anti-miR-181 b were studied using transwell and cell scarification test,respectively.Results The expression of miR-181b in HCC was higher than cancer-adjacent tissues and normal liver tissues.The differences among them were significant.There was a correlation between the high expression of miR-181b and invasiveness and metastasis in HCC.The protein expression of TIMP-3 in HCC was significantly lower than normal liver tissues and cancer-adjacent tissues.Expression of miR-181b mRNA was detected in various HCC cell lines such as Hep3B,HepG2,Huh 7,SKHep-1,SNU182,SNU449 and hepatocyte,with the expression of miR-181b in SKHep-1 being the highest (P＜0.01).TIMP3-3UTR was low when the expression of miR-181b was high (P＜0.05).The invasion and migration abilities of SKHep-1 were significantly inhibited by anti-miR-181b (P＜0.05).Conclusion The data suggested that miR-181b promoted invasion and migration of SKHep-1 by down-regulating TIMP-3 in HCC.