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  • Original Article
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MicroRNA-21 (miR-21) post-transcriptionally downregulates tumor suppressor Pdcd4 and stimulates invasion, intravasation and metastasis in colorectal cancer

Oncogene volume 27, pages 2128–2136 (2008)Cite this article

Abstract

Tumor-suppressor Pdcd4 inhibits transformation and invasion and is downregulated in cancers. So far, it has not been studied as to whether miRNAs, suppressing target expression by binding to the 3′-UTR, regulate Pdcd4 or invasion. The present study was conducted to investigate the regulation of Pdcd4, and invasion/intra-vasation, by miRNAs. A bioinformatics search revealed a conserved target-site for miR-21 within the Pdcd4-3′-UTR at 228–249 nt. In 10 colorectal cell lines, an inverse correlation of miR-21 and Pdcd4-protein was observed. Transfection of Colo206f-cells with miR-21 significantly suppressed a luciferase-reporter containing the Pdcd4-3′-UTR, whereas transfection of RKO with anti-miR-21 increased activity of this construct. This was abolished when a construct mutated at the miR-21/nt228–249 target site was used instead. Anti-miR-21-transfected RKO cells showed an increase of Pdcd4-protein and reduced invasion. Moreover, these cells showed reduced intra-vasation and lung metastasis in a chicken–embryo–metastasis assay. In contrast, overexpression of miR-21 in Colo206f significantly reduced Pdcd4-protein amounts and increased invasion, while Pdcd4-mRNA was unaltered. Resected normal/tumor tissues of 22 colorectal cancer patients demonstrated an inverse correlation between miR-21 and Pdcd4-protein. This is the first study to show that Pdcd4 is negatively regulated by miR-21. Furthermore, it is the first report to demonstrate that miR-21 induces invasion/intravasation/metastasis.

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Acknowledgements

HA was supported by Alfried Krupp von Bohlen und Halbach Foundation, Essen, Wilhelm-Sander-Stiftung, Munich, Auguste-Schaedel-Dantscher-Stiftung, Garmisch, Germany, Dr Hella-Buehler-Foundation, Heidelberg, B Braun Foundation, Malsung, The Hector Foundation, Weinheim and Dr Ingrid-zu-Solms Foundation, Frankfurt, Germany. We thank Dr Laura Nelson for critical reading of the article. This publication contains parts of the dissertation of Irfan A Asangani performed in partial fulfilment of the requirements for the PhD at DKFZ/University-Heidelberg.

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Authors and Affiliations

  1. Department of Experimental Surgery and Molecular Oncology of Solid Tumors, Medical Faculty Mannheim, University of Heidelberg, and DKFZ Heidelberg, Mannheim, Germany

    I A Asangani, S A K Rasheed, D A Nikolova, J H Leupold & H Allgayer

  2. Gene Regulation Section, Basic Research Laboratory, National Cancer Institute, Frederick, MD, USA

    N H Colburn

  3. Department of Surgery, Medical Faculty Mannheim, University of Heidelberg, Mannheim, Germany

    S Post

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  1. I A Asangani
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  2. S A K Rasheed
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  3. D A Nikolova
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Correspondence to H Allgayer.

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

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Asangani, I., Rasheed, S., Nikolova, D. et al. MicroRNA-21 (miR-21) post-transcriptionally downregulates tumor suppressor Pdcd4 and stimulates invasion, intravasation and metastasis in colorectal cancer. Oncogene 27, 2128–2136 (2008). https://doi.org/10.1038/sj.onc.1210856

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