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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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References

  • Bartel DP . (2004). MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116: 281–297.

    Article  CAS  PubMed  Google Scholar 

  • Berezikov E, Guryev V, van de Belt J, Wienholds E, Plasterk RH, Cuppen E . (2005). Phylogenetic shadowing and computational identification of human microRNA genes. Cell 120: 21–24.

    Article  CAS  PubMed  Google Scholar 

  • Bitomsky N, Bohm M, Klempnauer KH . (2004). Transformation suppressor protein Pdcd4 interferes with JNK-mediated phosphorylation of c-Jun and recruitment of the coactivator p300 by c-Jun. Oncogene 23: 7484–7493.

    Article  CAS  PubMed  Google Scholar 

  • Calin GA, Dumitru CD, Shimizu M, Bichi R, Zupo S, Noch E et al. (2002). Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukaemia. Proc Natl Acad Sci USA 99: 15524–15529.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Chan JA, Krichevsky AM, Kosik KS . (2005). MicroRNA-21 is an antiapoptotic factor in human glioblastoma cells. Cancer Res 65: 6029–6033.

    Article  CAS  PubMed  Google Scholar 

  • Chen CZ, Li L, Lodish HF, Bartel DP . (2004). MicroRNAs modulate hematopoietic lineage differentiation. Science 303: 83–86.

    Article  CAS  PubMed  Google Scholar 

  • Chen Y, Knosel T, Kristiansen G, Pietas A, Garber ME, Matsuhashi S et al. (2003). Loss of PDCD4 expression in human lung cancer correlates with tumour progression and prognosis. Pathology 200: 640–646.

    Article  CAS  Google Scholar 

  • Cmarik J, Min H, Hegamyer G, Zhan S, Kulesz-Martin M, Yoshinaga H et al. (1999). Differentially expressed protein Pdcd4 inhibits tumor promoter-induced neoplastic transformation. Proc Natl Acad Sci USA 96: 14037–14042.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Croce CM, Calin GA . (2005). miRNAs, cancer, and stem cell division. Cell 122: 6–7.

    Article  CAS  PubMed  Google Scholar 

  • Doench JG, Sharp PA . (2004). Specificity of microRNA target selection in translational repression. Genes & Dev 18: 504–511.

    Article  CAS  Google Scholar 

  • Dorrello NV, Peschiaroli A, Guardavaccaro D, Colburn NH, Sherman NE, Pagano M . (2006). S6K1- and betaTRCP-mediated degradation of PDCD4 promotes protein translation and cell growth. Science 314: 467–471.

    Article  CAS  PubMed  Google Scholar 

  • Esquela-Kerscher A, Slack FJ . (2006). Oncomirs—microRNAs with a role in cancer. Nat Rev Cancer 6: 259–269.

    Article  CAS  PubMed  Google Scholar 

  • Göke R, Gregel C, Göke A, Arnold R, Schmidt H, Lankat-Buttgereit B . (2004). Programmed cell death protein 4 (PDCD4) acts as a tumor suppressor in neuroendocrine tumor cells. Ann NY Acad Sci 1014: 220–222.

    Article  PubMed  Google Scholar 

  • Hammond SM . (2006). MicroRNAs as oncogenes. Curr Opin Genet Dev 16: 4–9.

    Article  CAS  PubMed  Google Scholar 

  • Iorio MV, Ferracin M, Liu CG, Veronese A, Spizzo R, Sabbioni S et al. (2005). MicroRNA gene expression deregulation in human breast cancer. Cancer Res 65: 7065–7070.

    Article  CAS  PubMed  Google Scholar 

  • Jansen AP, Camalier C, Colburn NH . (2005). Epidermal expression of the translation inhibitor programmed cell death 4 suppresses tumorigenesis. Cancer Res 65: 6034–6041.

    Article  CAS  PubMed  Google Scholar 

  • Kang M, Ahn HS, Lee JY, Matsuhashi S, Park WY . (2002). Up-regulation of PDCD4 in senescent human diploid fibroblasts. Biochem Biophys Res Commun 293: 617–621.

    Article  CAS  PubMed  Google Scholar 

  • Lankat-Buttgereit B, Gregel C, Knolle A, Hasilik A, Arnold R, Goke R . (2004). Pdcd4 inhibits growth of tumor cells by suppression of carbonic anhydrase type II. Mol Cell Endocrinol 214: 149–153.

    Article  CAS  PubMed  Google Scholar 

  • Leupold JH, Yang H-S, Colburn NH, Asangani I, Post S, Allgayer H . (2007). Tumor suppressor Pdcd4 inhibits invasion/intravasation and regulates urokinase receptor (u-PAR) gene expression via Sp-transcription factors. Oncogene 26: 4550–4562.

    Article  CAS  PubMed  Google Scholar 

  • Lu J, Getz G, Miska EA, Alvarez-Saavedra E, Lamb J, Peck D et al. (2005). MicroRNA expression profiles classify human cancers. Nature 425: 834–838.

    Article  Google Scholar 

  • Meister G, Landthaler M, Dorsett Y, Tuschl T . (2004). Sequence-specific inhibition of microRNA and siRNA-induced RNA silencing. RNA 10: 544–550.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Meng F, Henson R, Lang M, Wehbe H, Maheshwari S, Mendell JT et al. (2006). Involvement of human micro-RNA in growth and response to chemotherapy in human cholangiocarcinoma cell lines. Gastroenterology 130: 2113–2129.

    Article  CAS  PubMed  Google Scholar 

  • Mudduluru G, Medved F, Grobholz R, Jost C, Gruber A, Leupold JH et al. (2007). Loss of programmed cell death 4 expression marks adenoma-carcinoma transition, correlates inversely with phosphorylated protein kinase B, and is an independent prognostic factor in resected colorectal cancer. Cancer 110: 1697–1707.

    Article  CAS  PubMed  Google Scholar 

  • Onishi Y, Hashimoto S, Kizaki H . (1998). Cloning of the TIS gene suppressed by topoisomerase inhibitors. Gene 215: 453–459.

    Article  CAS  PubMed  Google Scholar 

  • Palamarchuk A, Efanov A, Maximov V, Aqeilan RI, Croce CM, Pekarsky Y . (2005). Akt Phosphorylates and regulates Pdcd4 tumor suppressor protein. Cancer Res 65: 11282–11286.

    Article  CAS  PubMed  Google Scholar 

  • Pfaffl MW . (2001). A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29: e45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pillai RS, Bhattacharyya SN, Artus CG, Zoller T, Cougot N, Basyuk E et al. (2005). Inhibition of translational initiation by let-7 microRNA in human cells. Science 309: 1573–1576.

    Article  CAS  PubMed  Google Scholar 

  • Roldo C, Missiaglia E, Hagan JP, Falconi M, Capelli P, Bersani S et al. (2006). MicroRNA expression abnormalities in pancreatic endocrine and acinar tumors are associated with distinctive pathologic features and clinical behavior. J Clin Oncol 24: 4677–4684.

    Article  CAS  PubMed  Google Scholar 

  • Schlichter U, Burk O, Worpenberg S, Klempnauer KH . (2001). The chicken Pdcd4 gene is regulated by v-Myb. Oncogene 20: 669–676.

    Article  Google Scholar 

  • Shibahara K, Asano M, Ishida Y, Aoki T, Koike T, Honjo T . (1995). Isolation of a novel mouse gene MA-3 that is induced upon programmed cell death. Gene 166: 297–301.

    Article  CAS  PubMed  Google Scholar 

  • Si M-L, Zhu S, Wu H, Lu Z, Wu F, Mo YY . (2007). miR-21-mediated tumor growth. Oncogene 26: 2799–2803.

    Article  CAS  PubMed  Google Scholar 

  • Yang H, Cho M, Zakowicz H, Hegamyer G, Sonenberg N, Colburn NH . (2004). A Novel function of the MA-3 domains in transformation and translation suppressor Pdcd4 is essential for its binding to eukaryotic translation initiation factor 4A. Mol Cell Bio 24: 3894–3906.

    Article  CAS  Google Scholar 

  • Yang H, Knies J, Stark C, Colburn NH . (2003). Pdcd4 suppresses tumor phenotype in JB6 cells by inhibiting AP-1 transactivation. Oncogene 22: 3712–3720.

    Article  CAS  PubMed  Google Scholar 

  • Yang H, Matthews C, Clair T, Wang Q, Baker AR, Li CC et al. (2006). Tumorigenesis suppressor Pdcd4 downregulates MAP4K1 expression to suppress colon carcinoma cell invasion. Mol Cell Biol 26: 1297–1306.

    Article  PubMed  PubMed Central  Google Scholar 

  • Zakowicz H, Yang H, Stark C, Wlodawer A, Laronde-Leblanc N, Colburn NH . (2005). Mutational analysis of the DEAD-box RNA helicase eIF4AII characterizes its interaction with transformation suppressor Pdcd4 and eIF4GI. RNA 11: 261–274.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zamore PD, Haley B . (2005). Ribo-gnome: the big world of small RNAs. Science 309: 1519–1524.

    Article  CAS  PubMed  Google Scholar 

  • Zhang Z, DuBois R . (2001). Detection of differentially expressed genes in human colon carcinoma cells treated with a selective COX-2 inhibitor. Oncogene 20: 4450–4456.

    Article  CAS  PubMed  Google Scholar 

  • Zhu S, Si M-L, Wu H, Mo YY . (2007). MicroRNA-21 targets the tumor suppressor gene tropomyosin 1 (TPM1). J Biol Chem 282: 14328–14336.

    Article  CAS  PubMed  Google Scholar 

  • Zijlstra A, Mellor R, Panzarella G, Aimes RT, Hooper JD, Marchenko ND et al. (2002). A quantitative analysis of rate-limiting steps in the metastatic cascade using human-specific real-time polymerase chain reaction. Cancer Res 62: 7083–7092.

    CAS  PubMed  Google Scholar 

Download references

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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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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