摘要
Objective To explore the role and regulation of guanine nucleotide-binding protein G（i）, a-1 subunit （GNAI1） in hepatocellular carcinoma （HCC）. Methods Expression of GNAI1 in HCC samples was determined by qRT-PCR and immunohistochemical （IHC） staining. Huh-7 and SNU-387 cells stably expressing GNAI1 were established by the infection of lentivirus transducing unit containing GNAI1. siRNA against GNAI1 was transfected into SMMC-7721 cells to knock down the GNAI1 expression in HCC cells. Mir-320a/c/d mimics were transfected into SMMC-7721 and SK-Hep-1 cells and the expression of GNAll was determined by Western blot. The migration and invasion of Huh-7, SNU-387, SK-Hep-1 and SMMC-7721 cells were investigated by Transwell assays. Results The GNAI1 protein was significantly downregulated in HCC samples without changes in its mRNA levels. GNAI1 could inhibit the migration and invasion of HCC cells in vitro. Further investigations indicated that GNAI1 was a target of miR-320a/c/d in HCC cells. Transwell assays demonstrated that these microRNAs could promote the migratory ability and invasivesess of HCC cells in vitro. Conclusions GNAII is downregulated in HCC and inhibits the migration and invasion of HCC cells. This study is the first to investigate the role of GNAI1 in cancer. Regulation of GNAI1 by miR-320a/c/d indicates new therapeutic avenues for targeting HCC metastasis.
Objective To explore the role and regulation of guanine nucleotide-binding protein G(i),α-1 subunit(GNAI1) in hepatocellular carcinoma(HCC). Methods Expression of GNAI1 in HCC samples was determined by qRT-PCR and immunohistochemical(IHC) staining.Huh-7 and SNU-387 cells stably expressing GNAI1 were established by the infection of lentivirus transducing unit containing GNAI1.siRNA against GNAI1 was transfected into SMMC-7721 cells to knock down the GNAI1 expression in HCC cells.Mir-320a/c/d mimics were transfected into SMMC-7721 and SK-Hep-1 cells and the expression of GNAI1 was determined by Western blot.The migration and invasion of Huh-7,SNU-387,SK-Hep-1 and SMMC-7721 cells were investigated by Transwell assays. Results The GNAI1 protein was significantly downregulated in HCC samples without changes in its mRNA levels.GNAI1 could inhibit the migration and invasion of HCC cells in vitro.Further investigations indicated that GNAI1 was a target of miR-320a/c/d in HCC cells.Transwell assays demonstrated that these microRNAs could promote the migratory ability and invasivesess of HCC cells in vitro. Conclusions GNAI1 is downregulated in HCC and inhibits the migration and invasion of HCC cells.This study is the first to investigate the role of GNAI1 in cancer.Regulation of GNAI1 by miR-320a/c/d indicates new therapeutic avenues for targeting HCC metastasis.
基金
supported by grants from the National 973 Key Basic Research Program(No.2011CB933100)
National Natural Science Foundation of China(No.81125016 and 81101481)
Science and Technology Commission of Shanghai Municipality(No.11XD1404500 and 10JC1414200)
Shanghai Health Bureau(No.XYQ2011047 and XBR2011039)
