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MicroRNA-21 promotes cell transformation by targeting the programmed cell death 4 gene

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Abstract

MicroRNAs (miRNAs) are small noncoding RNA molecules that negatively control expression of target genes in animals and plants. The microRNA-21 gene (mir-21) has been identified as the only miRNA commonly overexpressed in solid tumors of the lung, breast, stomach, prostate, colon, brain, head and neck, esophagus and pancreas. We initiated a screen to identify miR-21 target genes using a reporter assay and identified a potential miR-21 target in the 3′-UTR of the programmed cell death 4 (PDCD4) gene. We cloned the full-length 3′-UTR of human PDCD4 downstream of a reporter and found that mir-21 downregulated, whereas a modified antisense RNA to miR-21 upregulated reporter activity. Moreover, deletion of the putative miR-21-binding site (miRNA regulatory element, MRE) from the 3′-UTR of PDCD4, or mutations in the MRE abolished the ability of miR-21 to inhibit reporter activity, indicating that this MRE is a critical regulatory region. Western blotting showed that Pdcd4 protein levels were reduced by miR-21 in human and mouse cells, whereas quantitative real-time PCR revealed little difference at the mRNA level, suggesting translational regulation. Finally, overexpression of mir-21 in MCF-7 human breast cancer cells and mouse epidermal JB6 cells promoted soft agar colony formation by downregulating Pdcd4 protein levels. The demonstration that miR-21 promotes cell transformation supports the concept that mir-21 functions as an oncogene by a mechanism that involves translational repression of the tumor suppressor Pdcd4.

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Acknowledgements

YL is supported by the career development program and a pilot grant from the Center for Genomics and Integrated Biology at University of Louisville funded by NIEHS P30ES014443. This research was supported in part by NCI R21-CA124811 to CMK. ML is supported by PUJIANG program from the Committee of Shanghai Science and Technology (06PJ14105).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Louisville, Louisville, KY, USA

    Z Lu, V Stribinskis, C M Klinge, K S Ramos & Y Li

  2. Center for Genetics and Molecular Medicine, School of Medicine, University of Louisville, Louisville, KY, USA

    Z Lu, V Stribinskis, C M Klinge, K S Ramos & Y Li

  3. State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China

    M Liu

  4. Gene Regulation Section, Laboratory of Cancer Prevention, Center for Cancer Research, National Cancer Institute at Frederick, Frederick, MD, USA

    N H Colburn

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  1. Z Lu
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  2. M Liu
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  3. V Stribinskis
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  4. C M Klinge
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Correspondence to Y Li.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc)

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Lu, Z., Liu, M., Stribinskis, V. et al. MicroRNA-21 promotes cell transformation by targeting the programmed cell death 4 gene. Oncogene 27, 4373–4379 (2008). https://doi.org/10.1038/onc.2008.72

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  • DOI: https://doi.org/10.1038/onc.2008.72

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