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Growth-promoting and tumourigenic activity of c-Myc is suppressed by Hhex

Oncogene volume 34, pages 3011–3022 (2015)Cite this article

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Abstract

c-Myc transcription factor is a key protein involved in cellular growth, proliferation and metabolism. c-Myc is one of the most frequently activated oncogenes, highlighting the need to identify intracellular molecules that interact directly with c-Myc to suppress its function. Here we show that Hhex is able to interact with the basic region/helix-loop-helix/leucine zipper of c-Myc. Knockdown of Hhex increases proliferation rate in hepatocellular carcinoma cells, whereas Hhex expression cell-autonomously reduces cell proliferation rate in multiple cell lines by increasing G1 phase length through a c-Myc-dependent mechanism. Global transcriptomic analysis shows that Hhex counter-regulates multiple c-Myc targets involved in cell proliferation and metabolism. Concomitantly, Hhex expression leads to reduced cell size, lower levels of cellular RNA, downregulation of metabolism-related genes, decreased sensitivity to methotrexate and severe reduction in the ability to form tumours in nude mouse xenografts, all indicative of decreased c-Myc activity. Our data suggest that Hhex is a novel regulator of c-Myc function that limits c-Myc activity in transformed cells.

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Acknowledgements

We thank Dr Di Crocce for providing c-Myc truncated plasmids and Dr Ken Zaret for providing total RNA from Hhex-deficient E9.5 mouse embryos. We also thank Dr Laia Tolosa for assistance with Olympus ScanR screening station, Juan Jose Lozano for microarray analysis, Cristina Corchero for assistance in qRT-PCR and Ana Flores and Mª Jesus Seguí (Cell Culture and Cell Sorting facility at the SCSIE from Universidad de Valencia) for assistance in flow cytometry and cell sorting. VM and FS were recipients of pre-doctoral fellowships from the Ministerio de Ciencia e Innovacion. This work was supported by the Ministerio de Ciencia e Innovacion (grant number SAF2010-15376 and SAF2011-29718).

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  1. V Marfil and M Blazquez: These authors contributed equally to this work.

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  1. Unidad de Hepatología Experimental, CIBERehd, Instituto de Investigación Sanitaria La Fe, Avda. Fernando Abril Martorell 106, Valencia, Spain

    V Marfil, M Blazquez, F Serrano, J V Castell & R Bort

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Marfil, V., Blazquez, M., Serrano, F. et al. Growth-promoting and tumourigenic activity of c-Myc is suppressed by Hhex. Oncogene 34, 3011–3022 (2015). https://doi.org/10.1038/onc.2014.240

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