关键词: 肝细胞癌, miR?203a?3p, LASP1, 增殖, 迁移, 侵袭, 凋亡, Akt/GSK?3β/Snail信号通路

Abstract: Objective To investigate the effect of miR?203a?3p on the biological behavior of hepatocellular carcinoma (HCC) cells and its related molecular mechanism. Methods The miR?203a?3p mimics (miR?203a?3p mimics), negative control (miR?NC mimics), LIM and SH3 protein 1 (LASP1) overexpression plasmid (pcDNS?LASP1) and negative control plasmid (pcDNA?NC) were transfected into HepG2 cells. The expressions of miR?203a?3p and LASP1 mRNA were detected by qRT?PCR; cell proliferation was detected by the CCK?8 method and the allogeneic transplantation experiment; cell migration ability was detected by Wound healing assay; cell invasion ability was detected by Transwell; cell apoptosis was detected by Annexin V?FITC / PI method; Double luciferase reporter was used to detect the targeting relationship between miR?203a?3p and LASP1; Western blot was used to detect the protein expressions of LASP1, protein kinase B (Akt), phosphorylated Akt (p?Akt), glycogen synthase kinase?3β (GSK?3β), phosphorylated GSK?3β (p?GSK?3β) and Snail. Results Compared with those in the miR?NC group, the proliferation activity, migration rate, invasion number, LASP1 mRNA and protein expressions, the ratio of p?Akt/Akt and p?GSK?3β/GSK?3β, and the protein expression of Snail of HepG2 cells in the miR?203a?3p group were significantly decreased; the volume and mass of transplanted tumor in mice were significantly decreased;  the apoptosis rate was significantly increased (all P<0.01). Targetscan software prediction showed that miR?203a?3p had a targeting relationship with LASP1. Compared with that of LASP1?Wt+miR?NC group, the relative luciferase activity of LASP1?Wt+miR?203a?3p group was significantly decreased (P<0.001). Compared with those of the miR?203a?3p+pcDNA?NC group, the proliferation activity, migration rate, invasion number, p?Akt/Akt and p?GSK?3 β/GSK?3 β ratio, the protein expression of Snail protein of HepG2 cells in the miR?203a?3p+LASP1 group were significantly increased; the volume and mass of transplanted tumor in mice were also significantly increased; the apoptosis rate was significantly decreased (all P<0.01). Conclusions The miR?203a?3p may regulate the activity of Akt/GSK?3β/Snail signaling pathway by targeting the inhibition of LASP1 expression, thereby regulating the biological behavior of HCC cells.

Key words: Hepatocellular carcinoma, miR?203a?3p, LASP1, Proliferation, Migration, Invasion, Apoptosis, Akt/GSK?3β/Snail signaling pathway