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Human TRIB2 is a repressor of FOXO that contributes to the malignant phenotype of melanoma cells

Oncogene volume 29, pages 2973–2982 (2010)Cite this article

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Abstract

FOXO transcription factors are evolutionarily conserved proteins that orchestrate gene expression programs known to control a variety of cellular processes such as cell cycle, apoptosis, DNA repair and protection from oxidative stress. As the abrogation of FOXO function is a key feature of many tumor cells, regulation of FOXO factors is receiving increasing attention in cancer research. In order to discover genes involved in the regulation of FOXO activity, we performed a large-scale RNA-mediated interference (RNAi) screen using cell-based reporter systems that monitor transcriptional activity and subcellular localization of FOXO. We identified genes previously implicated in phosphoinositide 3-kinase/Akt signaling events, which are known to be important for FOXO function. In addition, we discovered a previously unrecognized FOXO-repressor function of TRIB2, the mammalian homolog of the Drosophila gene tribbles. A cancer-profiling array revealed specific overexpression of TRIB2 in malignant melanoma, but not in other types of skin cancer. We provide experimental evidence that TRIB2 transcript levels correlate with the degree of cytoplasmic localization of FOXO3a. Moreover, we show that TRIB2 is important in the maintenance of the oncogenic properties of melanoma cells, as its silencing reduces cell proliferation, colony formation and wound healing. Tumor growth was also substantially reduced upon RNAi-mediated TRIB2 knockdown in an in vivo melanoma xenograft model. Our studies suggest that TRIB2 provides the melanoma cells with growth and survival advantages through the abrogation of FOXO function. Altogether, our results show the potential of large-scale cell-based RNAi screens to identify promising diagnostic markers and therapeutic targets.

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Acknowledgements

This work was supported by a grant from the Spanish MEC (project BIO2006-02432). FZ is the recipient of a Marie Curie Fellowship. We acknowledge the expert technical assistance of D Megias, J C Cigudosa, J Monsech and O Dominguez. We thank M Hu, T Unterman and E Kiss-Toth for providing plasmids and JF Martinez for helpful discussions and critical reading of this paper. We are indebted to R Bernards for advice and for providing us with the NKI shRNA library.

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Authors and Affiliations

  1. Experimental Therapeutics Program, Centro Nacional de Investigaciones Oncologicas (CNIO), Madrid, Spain

    F Zanella, O Renner, B García, S Peregrina, A Carnero & W Link

  2. Genomics Unit, Centro Nacional de Investigaciones Cardiovasculares (CNIC), Madrid, Spain

    S Callejas & A Dopazo

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  1. F Zanella
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  2. O Renner
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  3. B García
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  4. S Callejas
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Correspondence to W Link.

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Zanella, F., Renner, O., García, B. et al. Human TRIB2 is a repressor of FOXO that contributes to the malignant phenotype of melanoma cells. Oncogene 29, 2973–2982 (2010). https://doi.org/10.1038/onc.2010.58

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