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. 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 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. Mer-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 a ssays demonstrated that these microRNAs could promote the migratory ability and invasive sess of HCC cells in vitro. Conclusions GNAI1 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.