MicroRNA/gene profiling unveils early molecular changes and nuclear factor erythroid related factor 2 (NRF2) activation in a rat model recapitulating human hepatocellular carcinoma (HCC)

Authors

  • Annalisa Petrelli,

    1. IRCC, Institute for Cancer Research and Treatment, University of Torino School of Medicine, Torino, Italy
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    • These authors equally contributed to the work.

  • Andrea Perra,

    1. Department of Biomedical Sciences, Unit of Oncology and Molecular Pathology, University of Cagliari, Cagliari, Italy
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    • These authors equally contributed to the work.

  • Davide Cora,

    1. IRCC, Institute for Cancer Research and Treatment, University of Torino School of Medicine, Torino, Italy
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    • These authors equally contributed to the work.

  • Pia Sulas,

    1. Department of Biomedical Sciences, Unit of Oncology and Molecular Pathology, University of Cagliari, Cagliari, Italy
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  • Silvia Menegon,

    1. IRCC, Institute for Cancer Research and Treatment, University of Torino School of Medicine, Torino, Italy
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  • Claudia Manca,

    1. Department of Biomedical Sciences, Unit of Oncology and Molecular Pathology, University of Cagliari, Cagliari, Italy
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  • Cristina Migliore,

    1. IRCC, Institute for Cancer Research and Treatment, University of Torino School of Medicine, Torino, Italy
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  • Marta Anna Kowalik,

    1. Department of Biomedical Sciences, Unit of Oncology and Molecular Pathology, University of Cagliari, Cagliari, Italy
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  • Giovanna Maria Ledda-Columbano,

    1. Department of Biomedical Sciences, Unit of Oncology and Molecular Pathology, University of Cagliari, Cagliari, Italy
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  • Silvia Giordano,

    Corresponding author
    1. IRCC, Institute for Cancer Research and Treatment, University of Torino School of Medicine, Torino, Italy
    • Address reprint requests to: Amedeo Columbano, Ph.D., Department of Biomedical Sciences, Unit of Oncology and Molecular Pathology, University of Cagliari, Via Porcell 4, 09124 Cagliari, Italy. E-mail: columbano@unica.it; fax: +39-070-666062. Silvia Giordano, M.D., Ph.D., Department of Oncology, University of Torino, Medical School, Institute for Cancer Research and Treatment (IRCC), Strada Provinciale 142, Candiolo, Torino, 10060, Italy. E-mail silvia.giordano@unito.it; fax +39 011 9933225.

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    • These authors equally contributed to the work.

  • Amedeo Columbano

    Corresponding author
    1. Department of Biomedical Sciences, Unit of Oncology and Molecular Pathology, University of Cagliari, Cagliari, Italy
    • Address reprint requests to: Amedeo Columbano, Ph.D., Department of Biomedical Sciences, Unit of Oncology and Molecular Pathology, University of Cagliari, Via Porcell 4, 09124 Cagliari, Italy. E-mail: columbano@unica.it; fax: +39-070-666062. Silvia Giordano, M.D., Ph.D., Department of Oncology, University of Torino, Medical School, Institute for Cancer Research and Treatment (IRCC), Strada Provinciale 142, Candiolo, Torino, 10060, Italy. E-mail silvia.giordano@unito.it; fax +39 011 9933225.

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    • These authors equally contributed to the work.


  • Potential conflict of interest: Nothing to report.

  • Supported by Associazione Italiana Ricerca sul Cancro (AIRC, Grants IG-11821 to A.C. and IG-11819 to S.G.), Ministero Università e Ricerca Scientifica (PRIN 2009X23L78 to S.G. and PRIN 2010LC747T to A.C.), R.A.S. 2012 to A.C. C.M. is an AIRC fellow. M.A.K. is a FIRC fellow.

Abstract

Studies on gene and/or microRNA (miRNA) dysregulation in the early stages of hepatocarcinogenesis are hampered by the difficulty of diagnosing early lesions in humans. Experimental models recapitulating human hepatocellular carcinoma (HCC) are then used to perform this analysis. We performed miRNA and gene expression profiling to characterize the molecular events involved in the multistep process of hepatocarcinogenesis in the resistant-hepatocyte rat model. A high percentage of dysregulated miRNAs/genes in HCC were similarly altered in early preneoplastic lesions positive for the stem/progenitor cell marker cytokeratin-19, indicating that several HCC-associated alterations occur from the very beginning of the carcinogenic process. Our analysis also identified miRNA/gene-target networks aberrantly activated at the initial stage of hepatocarcinogenesis. Activation of the nuclear factor erythroid related factor 2 (NRF2) pathway and up-regulation of the miR-200 family were among the most prominent changes. The relevance of these alterations in the development of HCC was confirmed by the observation that NRF2 silencing impaired while miR-200a overexpression promoted HCC cell proliferation in vitro. Moreover, T3-induced in vivo inhibition of the NRF2 pathway accompanied the regression of cytokeratin-19-positive nodules, suggesting that activation of this transcription factor contributes to the onset and progression of preneoplastic lesions towards malignancy. The finding that 78% of genes and 57% of dysregulated miRNAs in rat HCC have been previously associated with human HCC as well underlines the translational value of our results. Conclusion: This study indicates that most of the molecular changes found in HCC occur in the very early stages of hepatocarcinogenesis. Among these, the NRF2 pathway plays a relevant role and may represent a new therapeutic target. (Hepatology 2014;58:228–241)

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