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

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          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.

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          Most cited references17

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          Diagnostic and prognostic microRNAs in stage II colon cancer.

          Troels Schepeler, Jørgen T Reinert, Marie Ostenfeld … (2008)
          MicroRNAs (miRNA) are a class of small noncoding RNAs with important posttranscriptional regulatory functions. Recent data suggest that miRNAs are aberrantly expressed in many human cancers and that they may play significant roles in carcinogenesis. Here, we used microarrays to profile the expression of 315 human miRNAs in 10 normal mucosa samples and 49 stage II colon cancers differing with regard to microsatellite status and recurrence of disease. Several miRNAs were differentially expressed between normal tissue and tumor microsatellite subtypes, with miR-145 showing the lowest expression in cancer relative to normal tissue. Microsatellite status for the majority of cancers could be correctly predicted based on miRNA expression profiles. Furthermore, a biomarker based on miRNA expression profiles could predict recurrence of disease with an overall performance accuracy of 81%, indicating a potential role of miRNAs in determining tumor aggressiveness. The expression levels of miR-320 and miR-498, both included in the predictive biomarker, correlated with the probability of recurrence-free survival by multivariate analysis. We successfully verified the expression of selected miRNAs using real-time reverse transcription-PCR assays for mature miRNAs, whereas in situ hybridization was used to detect the accumulation of miR-145 and miR-320 in normal epithelial cells and adenocarcinoma cells. Functional studies showed that miR-145 potently suppressed growth of three different colon carcinoma cell lines. In conclusion, our results suggest that perturbed expression of numerous miRNAs in colon cancer may have a functional effect on tumor cell behavior, and, furthermore, that some miRNAs with prognostic potential could be of clinical importance.
          Article 0 comments Cited 153 times – based on 0 reviews     Review now
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            Multiple microRNAs modulate p21Cip1/Waf1 expression by directly targeting its 3' untranslated region.

            Lu T. Liu, L Liang, Justin Huang … (2010)
            Cyclin-dependent kinase inhibitor 1A (CDKN1A), also known as p21Cip1/Waf1, is a master downstream effector of tumor suppressors. In this study, we experimentally demonstrate through a high-throughput luciferase reporter screen that p21Cip1/Waf1 can be directly targeted by nearly 28 microRNAs (miRNAs). The results were further confirmed by a series of mutational analyses and luciferase reporter assays. These 28 miRNAs can substantially inhibit p21Cip1/Waf1 expression, predominantly at translational level. Many of these miRNAs were upregulated in cancers and might serve as modulators of oncogenesis. Furthermore, 8 of these 28 p21-regulating miRNAs are located in the chromosome 19 miRNA cluster, the largest miRNA gene cluster in humans, and they can clearly promote cell proliferation and cell-cycle progression in choriocarcinoma cells. In conclusion, our screening strategy provides an alternative approach to uncovering miRNA modulators of an individual mRNA, and it has identified multiple miRNAs that can suppress p21Cip1/Waf1 expression by directly targeting its 3' untranslated region.
            Article 0 comments Cited 150 times – based on 0 reviews     Review now
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              The role of microRNA expression pattern in human intrahepatic cholangiocarcinoma.

              Lei Chen, He-Xin Yan, Wen Yang … (2009)
              MicroRNAs are a small non-coding family of genes involved in the regulation of gene expression in a post-transcriptional manner and contribute to cell proliferation, differentiation and apoptosis. Our aims were to identify statistically unique miRNA profiles in human intrahepatic cholangiocarcinoma for diagnosis and investigate their specific involvement in various cell biological processes in cholangiocarcinoma. Laser capture microdissection techniques and TaqMan miRNA assays for mature miRNAs were performed to assess the genomewide expression of miRNAs in 27 human ICCs, 10 normal cholangiocyte cells and 8 normal liver tissues precisely and quantitatively. Two selected miRNAs, mir-204 and mir-320, were introduced into cholangiocarcinoma cell lines to examine their effects on potential target genes, Bcl-2 and Mcl-1, respectively. A cluster of 38 miRNAs was markedly distinguishable between tumor and normal tissues. At least two distinct clusters of tumor samples could be identified that were associated with the higher or lower expression levels of carbohydrate antigen 19-9. Moreover, the exogenous expression of mir-320 or mir-204 could negatively regulate Mcl-1 or Bcl-2 expression and facilitate chemotherapeutic drug-triggered apoptosis. miRNA expression profiles are closely associated with the biological and clinical behavior of ICC. The modulation of aberrantly expressed miRNAs might prove a promising therapeutic strategy.
              Article 0 comments Cited 89 times – based on 0 reviews     Review now
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                Author and article information

                Journal
                Journal ID (nlm-ta): Cancer Biol Med
                Journal ID (iso-abbrev): Cancer Biol Med
                Journal ID (publisher-id): CBM
                Title: Cancer Biology & Medicine
                Publisher: Chinese Anti-Cancer Association
                ISSN (Print): 2095-3941
                Publication date (Print): December 2012
                Volume: 9
                Issue: 4
                Pages: 234-241
                Affiliations
                [1 ]State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200032, China
                [2 ]State Key Laboratory for Diagnosis and Treatment for Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China
                Author notes
                [*]

                These authors contributed equally to this work.

                Correspondence to: Xiang-huo He

                E-mail: xhhe@ 123456shsci.org

                Article
                Publisher ID: cbm-09-04-234
                DOI: 10.7497/j.issn.2095-3941.2012.04.003
                PMC ID: 3643671
                PubMed ID: 23691483
                SO-VID: 2f8d645b-7879-49ea-8ed1-07a57a01087f
                Copyright © 2012 Cancer Biology & Medicine
                License:

                This work is licensed under a Creative Commons Attribution 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by/3.0/

                History
                Date received : 30 August 2012
                Date accepted : 07 November 2012
                Categories
                Subject: Original Article

                Keywords: gnai1,hcc,mir-320,migration,invasion
                Data availability:
                Keywords: gnai1, hcc, mir-320, migration, invasion

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