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The H19 endodermal enhancer is required for Igf2 activation and tumor formation in experimental liver carcinogenesis

Oncogene volume 19, pages 6376–6385 (2000)Cite this article

Abstract

The expression of the linked but reciprocally imprinted Igf2 and H19 genes is activated in adult liver in the course of tumor development. By in situ hybridization analysis we have shown that both the Igf2 and H19 RNAs are expressed in the majority of the neoplastic nodules, and that hepatocellular carcinomas are developed in an experimental model of liver carcinogenesis. H19 is also highly activated in smaller and less distinct hyperplastic regions. The few neoplastic areas showing Igf2 but no H19 RNA display loss of the maternally inherited allele at the Igf2/H19 locus. These data are compatible with the existence of a common activation mechanism of these two genes during liver carcinogenesis and with a stronger H19 induction in the pre-neoplastic lesions. By using mice carrying a deletion of the H19 endodermal enhancer, we show that this regulatory element is necessary for the activation of the Igf2 and H19 genes upon induction of liver carcinogenesis. Furthermore, multiple sites of the H19 endodermal enhancer region become hypersensitive to DNase I when the carcinogenesis process is induced. Lastly, liver tumors developed in mice paternally inheriting the H19 enhancer deletion are found to have marked growth delays, increased frequency of apoptotic nuclei, and lack of Igf2 mRNA expression, thus indicating that this regulatory element plays a major role in the progression of liver carcinogenesis, since it is required for the activation of the anti-apoptotic Igf2 gene.

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Acknowledgements

We are grateful to Shirley M Tilghman for providing the H19EnhΔ mice, Antonio Simeone for the invaluable directions on the in situ hybridization procedure, to M Bartolomei for the H19 3′ probe, to Enzo Cimini for the use of the cryostat and to Mario De Felice for the use of the microscope. We also thank Giovanni Sequino for excellent technical assistance in the animal house. This work was partially supported by grants from Associazione Italiana Ricerca sul Cancro and MURST 60% (to A Riccio). N Besnard was the recipient of fellowships from the Association pour la Recherche sur le Cancer (France) and the European Community.

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Authors and Affiliations

  1. Centro di Endocrinologia ed Oncologia Sperimentale ‘G Salvatore’, CNR,

    Maria Vernucci, Flavia Cerrato, Nathalie Besnard, Stefano Casola, Paolo V Pedone, Carmelo B Bruni & Andrea Riccio

  2. Dipartimento di Biologia e Patologia Cellulare e Molecolare ‘L Califano’, Università di Napoli ‘Federico II’, Napoli, Italy

    Maria Vernucci, Flavia Cerrato, Nathalie Besnard, Stefano Casola, Paolo V Pedone, Carmelo B Bruni & Andrea Riccio

  3. Dipartimento di Scienze Ambientali, Seconda Universitá di, Napoli, Italy

    Maria Vernucci, Paolo V Pedone & Andrea Riccio

  4. Centro di Biocristallografia, CNR, Napoli, Italy

    Flavia Cerrato

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  1. Maria Vernucci
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Vernucci, M., Cerrato, F., Besnard, N. et al. The H19 endodermal enhancer is required for Igf2 activation and tumor formation in experimental liver carcinogenesis. Oncogene 19, 6376–6385 (2000). https://doi.org/10.1038/sj.onc.1204024

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