A functional screening identifies five micrornas controlling glypican-3: role of mir-1271 down-regulation in hepatocellular carcinoma

Authors

  • Marion Maurel,

    1. Univ. Bordeaux, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
    2. INSERM, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
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  • Sandra Jalvy,

    1. Univ. Bordeaux, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
    2. INSERM, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
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    • These authors contributed equally to this work.

  • Yannick Ladeiro,

    1. Université Paris Descartes, Labex Immuno-oncology, Sorbonne Paris Cité, Faculté de Médecine, Paris, France
    2. INSERM, Altération génétique des tumeurs hépatiques, U674, IUH, F-75010 Paris, France
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    • These authors contributed equally to this work.

  • Chantal Combe,

    1. Univ. Bordeaux, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
    2. INSERM, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
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  • Laetitia Vachet,

    1. Univ. Bordeaux, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
    2. INSERM, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
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  • Francis Sagliocco,

    1. Univ. Bordeaux, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
    2. INSERM, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
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  • Paulette Bioulac-Sage,

    1. Univ. Bordeaux, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
    2. INSERM, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
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  • Vincent Pitard,

    1. Univ. Bordeaux, CIRID, UMR 5164, F-33000 Bordeaux, France
    2. CNRS, CIRID, UMR5164, F-33000 Bordeaux, France
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  • Hélène Jacquemin-Sablon,

    1. Univ. Bordeaux, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
    2. INSERM, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
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  • Jessica Zucman-Rossi,

    1. Université Paris Descartes, Labex Immuno-oncology, Sorbonne Paris Cité, Faculté de Médecine, Paris, France
    2. INSERM, Altération génétique des tumeurs hépatiques, U674, IUH, F-75010 Paris, France
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  • Benoît Laloo,

    1. Univ. Bordeaux, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
    2. INSERM, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
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  • Christophe F. Grosset

    Corresponding author
    1. Univ. Bordeaux, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
    2. INSERM, Physiopathologie du cancer du foie, U1053, F-33000 Bordeaux, France
    • Université Bordeaux Segalen, Physiopathologie du cancer du foie, INSERM U1053, 146 Rue Léo Saignat, F-33076 Bordeaux cedex, France
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    • fax: +33 (0)556 514 077


  • Potential conflict of interest: Nothing to report.

  • Supported by the Agence Nationale pour la Recherche (ANR)-Programme Jeunes Chercheurs (Paris, France) (Grant no: JC07_184264 to C.G.), by La Ligue Nationale Contre le Cancer, by the Biointelligence consortium (OSEO) and INCa (PAIR-CHC). M.M and Y.L. were recipients of fellowships from the Ministère de l'Enseignement Supérieur et de la Recherche (MESR) and ANRS, respectively.

Abstract

Hepatocellular carcinoma (HCC) is the major primary liver cancer. Glypican-3 (GPC3), one of the most abnormally expressed genes in HCC, participates in liver carcinogenesis. Based on data showing that GPC3 expression is posttranscriptionally altered in HCC cells compared to primary hepatocytes, we investigated the implication of microRNAs (miRNAs) in GPC3 overexpression and HCC. To identify GPC3-regulating miRNAs, we developed a dual-fluorescence FunREG (functional, integrated, and quantitative method to measure posttranscriptional regulations) system that allowed us to screen a library of 876 individual miRNAs. Expression of candidate miRNAs and that of GPC3 messenger RNA (mRNA) was measured in 21 nontumoral liver and 112 HCC samples. We then characterized the phenotypic consequences of modulating expression of one candidate miRNA in HuH7 cells and deciphered the molecular mechanism by which this miRNA controls the posttranscriptional regulation of GPC3. We identified five miRNAs targeting GPC3 3′-untranslated region (UTR) and regulating its expression about the 876 tested. Whereas miR-96 and its paralog miR-1271 repressed GPC3 expression, miR-129-1-3p, miR-1291, and miR-1303 had an inducible effect. We report that miR-1271 expression is down-regulated in HCC tumor samples and inversely correlates with GPC3 mRNA expression in a particular subgroup of HCC. We also report that miR-1271 inhibits the growth of HCC cells in a GPC3-dependent manner and induces cell death. Conclusion: Using a functional screen, we found that miR-96, miR-129-1-3p, miR-1271, miR-1291, and miR-1303 differentially control GPC3 expression in HCC cells. In a subgroup of HCC, the up-regulation of GPC3 was associated with a concomitant down-regulation of its repressor miR-1271. Therefore, we propose that GPC3 overexpression and its associated oncogenic effects are linked to the down-regulation of miR-1271 in HCC. (HEPATOLOGY 2013)

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