Limits...
GNAI1 Suppresses Tumor Cell Migration and Invasion and is Post-Transcriptionally Regulated by Mir-320a/c/d in Hepatocellular Carcinoma.

Yao J, Liang LH, Zhang Y, Ding J, Tian Q, Li JJ, He XH - Cancer Biol Med (2012)

Bottom Line: The GNAI1 protein was significantly downregulated in HCC samples without changes in its mRNA levels.GNAI1 is downregulated in HCC and inhibits the migration and invasion of HCC cells.Regulation of GNAI1 by miR-320a/c/d indicates new therapeutic avenues for targeting HCC metastasis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200032, China ; State Key Laboratory for Diagnosis and Treatment for Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus

GNAI1 can significantly inhibit the migration and metastasis of HCC cells. A: Western blot of GNAI1 protein levels in Huh-7 and SNU-387 cells infected with lentiviral constructs containing GNAI1 or lentiviral constructs; B: Transwell migration assays of Huh-7 and SNU-387 cells stably overexpressing GNAI1 or the mock control. The representative images and quantification of the five randomly selected fields are shown. Values are shown as mean±SEM, along with the statistical significance; C: Transwell invasion assays of Huh-7 or SNU-387 cells expressing GNAI1 or the mock control; D: Western blot of GNAI1 expression levels in SMMC-7721 cells transfected with GNAI1-siRNAs or negative control-siRNA; E: Transwell migration and invasion assays of SMMC-7721 transfected with GNAI1-siRNA or negative control-siRNA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3643671&req=5

f2: GNAI1 can significantly inhibit the migration and metastasis of HCC cells. A: Western blot of GNAI1 protein levels in Huh-7 and SNU-387 cells infected with lentiviral constructs containing GNAI1 or lentiviral constructs; B: Transwell migration assays of Huh-7 and SNU-387 cells stably overexpressing GNAI1 or the mock control. The representative images and quantification of the five randomly selected fields are shown. Values are shown as mean±SEM, along with the statistical significance; C: Transwell invasion assays of Huh-7 or SNU-387 cells expressing GNAI1 or the mock control; D: Western blot of GNAI1 expression levels in SMMC-7721 cells transfected with GNAI1-siRNAs or negative control-siRNA; E: Transwell migration and invasion assays of SMMC-7721 transfected with GNAI1-siRNA or negative control-siRNA.

Mentions: We had previously reported that GNAI2 functions as an inhibitor of HCC migration and invasion. GNAI1 may possess biological functions similar to GNAI2 in HCC cells because both GNAI1 and GNAI2 are members of the Gαi family. To explore these functions, we established stable HCC cell lines by infecting Huh-7 and SNU-387 cells with a lentiviral construct that contained either the complete ORF of the GNAI1 gene or the vector alone (Figure 2A). The enhanced GNAI1 expression did not affect the growth of HCC cells. However, transwell assays without Matrigel indicated that GNAI1 could significantly suppress the migration of HCC cells compared with the vector groups (Figure 2B). Furthermore, transwell assays with Matrigel demonstrated that the invasiveness of the HCC cells was dramatically inhibited by GNAI1 compared with that by the vector groups (Figure 2C). To confirm whether GNAI1 can inhibit the migration and invasion of HCC cells, we knocked down the endogenous GNAI1 expression in SMMC-7721 cells with siRNA (Figure 2D). Consistent with the overexpression transwell assays, the knockdown of the endogenous GNAI1 significantly enhanced HCC cell migration and invasion (Figure 2E). Taken together, these observations indicate that GNAI1 is a negative regulator of HCC migration and invasion, similar to its family member GNAI2.


GNAI1 Suppresses Tumor Cell Migration and Invasion and is Post-Transcriptionally Regulated by Mir-320a/c/d in Hepatocellular Carcinoma.

Yao J, Liang LH, Zhang Y, Ding J, Tian Q, Li JJ, He XH - Cancer Biol Med (2012)

GNAI1 can significantly inhibit the migration and metastasis of HCC cells. A: Western blot of GNAI1 protein levels in Huh-7 and SNU-387 cells infected with lentiviral constructs containing GNAI1 or lentiviral constructs; B: Transwell migration assays of Huh-7 and SNU-387 cells stably overexpressing GNAI1 or the mock control. The representative images and quantification of the five randomly selected fields are shown. Values are shown as mean±SEM, along with the statistical significance; C: Transwell invasion assays of Huh-7 or SNU-387 cells expressing GNAI1 or the mock control; D: Western blot of GNAI1 expression levels in SMMC-7721 cells transfected with GNAI1-siRNAs or negative control-siRNA; E: Transwell migration and invasion assays of SMMC-7721 transfected with GNAI1-siRNA or negative control-siRNA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3643671&req=5

f2: GNAI1 can significantly inhibit the migration and metastasis of HCC cells. A: Western blot of GNAI1 protein levels in Huh-7 and SNU-387 cells infected with lentiviral constructs containing GNAI1 or lentiviral constructs; B: Transwell migration assays of Huh-7 and SNU-387 cells stably overexpressing GNAI1 or the mock control. The representative images and quantification of the five randomly selected fields are shown. Values are shown as mean±SEM, along with the statistical significance; C: Transwell invasion assays of Huh-7 or SNU-387 cells expressing GNAI1 or the mock control; D: Western blot of GNAI1 expression levels in SMMC-7721 cells transfected with GNAI1-siRNAs or negative control-siRNA; E: Transwell migration and invasion assays of SMMC-7721 transfected with GNAI1-siRNA or negative control-siRNA.
Mentions: We had previously reported that GNAI2 functions as an inhibitor of HCC migration and invasion. GNAI1 may possess biological functions similar to GNAI2 in HCC cells because both GNAI1 and GNAI2 are members of the Gαi family. To explore these functions, we established stable HCC cell lines by infecting Huh-7 and SNU-387 cells with a lentiviral construct that contained either the complete ORF of the GNAI1 gene or the vector alone (Figure 2A). The enhanced GNAI1 expression did not affect the growth of HCC cells. However, transwell assays without Matrigel indicated that GNAI1 could significantly suppress the migration of HCC cells compared with the vector groups (Figure 2B). Furthermore, transwell assays with Matrigel demonstrated that the invasiveness of the HCC cells was dramatically inhibited by GNAI1 compared with that by the vector groups (Figure 2C). To confirm whether GNAI1 can inhibit the migration and invasion of HCC cells, we knocked down the endogenous GNAI1 expression in SMMC-7721 cells with siRNA (Figure 2D). Consistent with the overexpression transwell assays, the knockdown of the endogenous GNAI1 significantly enhanced HCC cell migration and invasion (Figure 2E). Taken together, these observations indicate that GNAI1 is a negative regulator of HCC migration and invasion, similar to its family member GNAI2.

Bottom Line: The GNAI1 protein was significantly downregulated in HCC samples without changes in its mRNA levels.GNAI1 is downregulated in HCC and inhibits the migration and invasion of HCC cells.Regulation of GNAI1 by miR-320a/c/d indicates new therapeutic avenues for targeting HCC metastasis.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200032, China ; State Key Laboratory for Diagnosis and Treatment for Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China.

ABSTRACT

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.

No MeSH data available.


Related in: MedlinePlus