MiR-101Inhibits Human Hepatocellular Carcinoma Cell Proliferation And Invasion By Regulating Girdin

Objective:Our previous studies have confirmed that the Girdin is highly expressed in hepatocellular carcinoma tissues, this study aims to explore why Girdin is highly expressed in hepatocellular carcinoma, and confirm that miR-101can specificity regulate the expression of Girdin and inhibit the ability of cell proliferation and invasion and metastasis of hepatocellular carcinoma.Methods:(1) qRT-PCR was used to detect the expression of Girdin mRNA in5HCC tissues and corresponding pericarcinomatous tissue;(2) The bioinformatics softwares for predicting miRNAs:miRanda and Targetscan were utilized to predict the interaction between miRNA and the3’-UTR of Girdin mRNA;(3) qRT-PCR was used to detect the expression of miR-101in HCC tissues and corresponding pericarcinomatous tissue;(4) qRT-PCR was used to detect the expression levels of Girdin mRNA in the HEPG2cells after transfection with miR-101;Western blot assay was used to detect the expression levels of Girdin protein in the HEPG2cells after transfection with miR-101;(5) We first build Girdin-3’UTR plasmid,then the plasmid and miR-101mimics transfection into HEPG2cells;Dual luciferase experiment was used to verify Girdin is a direct target gene of miR-101;(6) CCK was used to examine the effects of miR-101on the HEPG2cells proliferation;(7) Wound Healing assay was used to examine the effects of miR-101on the HEPG2cells migration;(8) Transwell was used to examine the effects of miR-101on the HEPG2cells invasion.Results:(1) The results showed that Girdin mRNA was significantly upregulated in HCC compared with adjacent normal tissues;(2) Targetscan and miRanda predicted Girdin mRNA was a target mRNA of miR-101;(3) The results showed that miR-101mRNA was significantly downregulated in HCC compared with adjacent normal tissues;(4) miR-101inhibits the expression levels of Girdin mRNA and protein in HEPG2cells;(5) Dual luciferase experiments show that miR-101transfection into HepG2cells can inhibit Girdin-3’UTR luciferase activity;(6) miR-101inhibits HEPG2cells proliferation;(7) miR-101inhibits HEPG2cells migration;(8) miR-101inhibits HEPG2cells invasion.Conclusion:(1) Girdin is highly expressed in hepatocellular carcinoma.The partly reason is that miR-101can directly regulate Girdin and miR-101is lowly expressed in hepatocellular carcinoma.(2)miR-101in hepatocellular carcinoma may by directly targeting Girdin to inhibit the ability of cell proliferation and invasion and metastasis of hepatocellular carcinoma.