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Attenuation of the beta-catenin/TCF4 complex in colorectal cancer cells induces several growth-suppressive microRNAs that target cancer promoting genes

Oncogene volume 31, pages 2750–2760 (2012)Cite this article

Abstract

Aberrant activation of the Wnt signaling pathway is causally involved in the formation of most colorectal cancers (CRCs). Although detailed knowledge exists regarding Wnt-regulated protein-coding genes, much less is known about the possible involvement of non-coding RNAs. Here we used TaqMan Array MicroRNA Cards, capable of detecting 664 unique human microRNAs (miRNAs), to describe changes of the miRNA transcriptome following disruption of beta-catenin/TCF4 activity in DLD1 CRC cells. Most miRNAs appeared to respond independent of host gene regulation and proximal TCF4 chromatin occupancy as inferred from expression microarray and ChIP–chip data. A module of miRNAs induced by abrogated Wnt signaling in vitro was downregulated in two independent series of human primary CRCs (n=76) relative to normal adjacent mucosa (n=34). Several of these miRNAs (miR-145, miR-126, miR-30e-3p and miR-139-5p) markedly inhibited CRC cell growth in vitro when ectopically expressed. By using an integrative approach of proteomics and expression microarrays, we found numerous mRNAs and proteins to be affected by ectopic miR-30e-3p levels. This included HELZ and PIK3C2A that were directly repressed by several miRNA binding sites as confirmed by luciferase reporter assays in combination with mutational analyses. Finally, small interfering RNA-mediated downregulation of PIK3C2A, but not HELZ, was sufficient on its own to restrict CRC cell growth. Collectively, our study demonstrates that multiple miRNAs are upregulated as a consequence of forced attenuation of Wnt signaling in CRC cells, and some of these miRNAs inhibit cell growth with concomitant suppression of several growth-stimulatory cancer-related genes.

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References

  • Baek D, Villen J, Shin C, Camargo FD, Gygi SP, Bartel DP . (2008). The impact of microRNAs on protein output. Nature 455: 64–71.

    Article  CAS  Google Scholar 

  • Blauwkamp TA, Chang MV, Cadigan KM . (2008). Novel TCF-binding sites specify transcriptional repression by Wnt signalling. EMBO J 27: 1436–1446.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Brennecke J, Hipfner DR, Stark A, Russell RB, Cohen SM . (2003). bantam encodes a developmentally regulated microRNA that controls cell proliferation and regulates the proapoptotic gene hid in Drosophila. Cell 113: 25–36.

    Article  CAS  Google Scholar 

  • Chen F, Zhu HH, Zhou LF, Wu SS, Wang J, Chen Z . (2010). Inhibition of c-FLIP expression by miR-512-3p contributes to Taxol-induced apoptosis in hepatocellular carcinoma cells. Oncol Rep 23: 1457–1462.

    CAS  PubMed  Google Scholar 

  • Chi SW, Zang JB, Mele A, Darnell RB . (2009). Argonaute HITS-CLIP decodes microRNA-mRNA interaction maps. Nature 460: 479–486.

    Article  CAS  Google Scholar 

  • Croce CM . (2009). Causes and consequences of microRNA dysregulation in cancer. Nat Rev Genet 10: 704–714.

    Article  CAS  Google Scholar 

  • Cselenyi CS, Lee E . (2008). Context-dependent activation or inhibition of Wnt-beta-catenin signaling by Kremen. Sci Signal 1: pe10.

    Article  Google Scholar 

  • Dehner M, Hadjihannas M, Weiske J, Huber O, Behrens J . (2008). Wnt signaling inhibits Forkhead box O3a-induced transcription and apoptosis through up-regulation of serum- and glucocorticoid-inducible kinase 1. J Biol Chem 283: 19201–19210.

    Article  CAS  Google Scholar 

  • Ding L, Xu Y, Zhang W, Deng Y, Si M, Du Y et al. (2010). MiR-375 frequently downregulated in gastric cancer inhibits cell proliferation by targeting JAK2. Cell Res 20: 784–793.

    Article  CAS  Google Scholar 

  • Elis W, Triantafellow E, Wolters NM, Sian KR, Caponigro G, Borawski J et al. (2008). Down-regulation of class II phosphoinositide 3-kinase alpha expression below a critical threshold induces apoptotic cell death. Mol Cancer Res 6: 614–623.

    Article  CAS  Google Scholar 

  • Filipowicz W, Bhattacharyya SN, Sonenberg N . (2008). Mechanisms of post-transcriptional regulation by microRNAs: are the answers in sight? Nat Rev Genet 9: 102–114.

    Article  CAS  Google Scholar 

  • Fullwood MJ, Liu MH, Pan YF, Liu J, Xu H, Mohamed YB et al. (2009). An oestrogen-receptor-alpha-bound human chromatin interactome. Nature 462: 58–64.

    Article  CAS  Google Scholar 

  • Gregorieff A, Pinto D, Begthel H, Destree O, Kielman M, Clevers H . (2005). Expression pattern of Wnt signaling components in the adult intestine. Gastroenterology 129: 626–638.

    Article  CAS  Google Scholar 

  • Griffiths-Jones S, Saini HK, van Dongen S, Enright AJ . (2008). miRBase: tools for microRNA genomics. Nucleic Acids Res 36: D154–D158.

    Article  CAS  Google Scholar 

  • Guo C, Sah JF, Beard L, Willson JK, Markowitz SD, Guda K . (2008). The noncoding RNA, miR-126, suppresses the growth of neoplastic cells by targeting phosphatidylinositol 3-kinase signaling and is frequently lost in colon cancers. Genes Chromosomes Cancer 47: 939–946.

    Article  CAS  Google Scholar 

  • Hamamoto R, Furukawa Y, Morita M, Iimura Y, Silva FP, Li M et al. (2004). SMYD3 encodes a histone methyltransferase involved in the proliferation of cancer cells. Nat Cell Biol 6: 731–740.

    Article  CAS  Google Scholar 

  • Hatzis P, van der Flier LG, van Driel MA, Guryev V, Nielsen F, Denissov S et al. (2008). Genome-wide pattern of TCF7L2/TCF4 chromatin occupancy in colorectal cancer cells. Mol Cell Biol 28: 2732–2744.

    Article  CAS  Google Scholar 

  • Heintzman ND, Ren B . (2009). Finding distal regulatory elements in the human genome. Curr Opin Genet Dev 19: 541–549.

    Article  CAS  Google Scholar 

  • Henson BJ, Bhattacharjee S, O'Dee DM, Feingold E, Gollin SM . (2009). Decreased expression of miR-125b and miR-100 in oral cancer cells contributes to malignancy. Genes Chromosomes Cancer 48: 569–582.

    Article  CAS  Google Scholar 

  • Herranz H, Perez L, Martin FA, Milan M . (2008). A wingless and notch double-repression mechanism regulates G1-S transition in the Drosophila wing. EMBO J 27: 1633–1645.

    Article  CAS  Google Scholar 

  • Ichimi T, Enokida H, Okuno Y, Kunimoto R, Chiyomaru T, Kawamoto K et al. (2009). Identification of novel microRNA targets based on microRNA signatures in bladder cancer. Int J Cancer 125: 345–352.

    Article  CAS  Google Scholar 

  • Kapinas K, Kessler CB, Delany AM . (2009). miR-29 suppression of osteonectin in osteoblasts: regulation during differentiation and by canonical Wnt signaling. J Cell Biochem 108: 216–224.

    Article  CAS  Google Scholar 

  • Kapinas K, Kessler CB, Ricks T, Gronowicz G, Delany AM . (2010). miR-29 modulates Wnt signaling in human osteoblasts through a positive feedback loop. J Biol Chem. 285: 25221–25231.

    Article  CAS  Google Scholar 

  • Kato M, Putta S, Wang M, Yuan H, Lanting L, Nair I et al. (2009). TGF-beta activates Akt kinase through a microRNA-dependent amplifying circuit targeting PTEN. Nat Cell Biol 11: 881–889.

    Article  CAS  Google Scholar 

  • Labbe E, Lock L, Letamendia A, Gorska AE, Gryfe R, Gallinger S et al. (2007). Transcriptional cooperation between the transforming growth factor-beta and Wnt pathways in mammary and intestinal tumorigenesis. Cancer Res 67: 75–84.

    Article  CAS  Google Scholar 

  • Li H, Ray G, Yoo BH, Erdogan M, Rosen KV . (2009). Down-regulation of death-associated protein kinase-2 is required for beta-catenin-induced anoikis resistance of malignant epithelial cells. J Biol Chem 284: 2012–2022.

    Article  CAS  Google Scholar 

  • Liao Y, Lonnerdal B . (2010). Beta-catenin/TCF4 transactivates miR-30e during intestinal cell differentiation. Cell Mol Life Sci. 67: 2969–2978.

    Article  CAS  Google Scholar 

  • Martello G, Zacchigna L, Inui M, Montagner M, Adorno M, Mamidi A et al. (2007). MicroRNA control of Nodal signalling. Nature 449: 183–188.

    Article  CAS  Google Scholar 

  • Medici D, Hay ED, Olsen BR . (2008). Snail and Slug promote epithelial-mesenchymal transition through beta-catenin-T-cell factor-4-dependent expression of transforming growth factor-beta3. Mol Biol Cell 19: 4875–4887.

    Article  CAS  Google Scholar 

  • Nagel R, le Sage C, Diosdado B, van der Waal M, Oude Vrielink JA, Bolijn A et al. (2008). Regulation of the adenomatous polyposis coli gene by the miR-135 family in colorectal cancer. Cancer Res 68: 5795–5802.

    Article  CAS  Google Scholar 

  • Ng SK, Neo SY, Yap YW, Karuturi RK, Loh ES, Liau KH et al. (2009). Ablation of phosphoinositide-3-kinase class II alpha suppresses hepatoma cell proliferation. Biochem Biophys Res Commun 387: 310–315.

    Article  CAS  Google Scholar 

  • Noonan EJ, Place RF, Pookot D, Basak S, Whitson JM, Hirata H et al. (2009). miR-449a targets HDAC-1 and induces growth arrest in prostate cancer. Oncogene 28: 1714–1724.

    Article  CAS  Google Scholar 

  • Ostenfeld MS, Bramsen JB, Lamy P, Villadsen SB, Fristrup N, Sorensen KD et al. (2010). miR-145 induces caspase-dependent and -independent cell death in urothelial cancer cell lines with targeting of an expression signature present in Ta bladder tumors. Oncogene 29: 1073–1084.

    Article  CAS  Google Scholar 

  • Railo A, Pajunen A, Itaranta P, Naillat F, Vuoristo J, Kilpelainen P et al. (2009). Genomic response to Wnt signalling is highly context-dependent–Evidence from DNA microarray and chromatin immunoprecipitation screens of Wnt/TCF targets. Exp Cell Res 315: 2690–2704.

    Article  CAS  Google Scholar 

  • Samuels Y, Diaz Jr LA., Schmidt-Kittler O, Cummins JM, Delong L, Cheong I et al. (2005). Mutant PIK3CA promotes cell growth and invasion of human cancer cells. Cancer Cell 7: 561–573.

    Article  CAS  Google Scholar 

  • Saydam O, Shen Y, Wurdinger T, Senol O, Boke E, James MF et al. (2009). Downregulated microRNA-200a in meningiomas promotes tumor growth by reducing E-cadherin and activating the Wnt/beta-catenin signaling pathway. Mol Cell Biol 29: 5923–5940.

    Article  CAS  Google Scholar 

  • Schepeler T, Reinert JT, Ostenfeld MS, Christensen LL, Silahtaroglu AN, Dyrskjot L et al. (2008). Diagnostic and prognostic microRNAs in stage II colon cancer. Cancer Res 68: 6416–6424.

    Article  CAS  Google Scholar 

  • Schneikert J, Behrens J . (2007). The canonical Wnt signalling pathway and its APC partner in colon cancer development. Gut 56: 417–425.

    Article  CAS  Google Scholar 

  • Selbach M, Schwanhausser B, Thierfelder N, Fang Z, Khanin R, Rajewsky N . (2008). Widespread changes in protein synthesis induced by microRNAs. Nature 455: 58–63.

    Article  CAS  Google Scholar 

  • Slaby O, Svoboda M, Michalek J, Vyzula R . (2009). MicroRNAs in colorectal cancer: translation of molecular biology into clinical application. Mol Cancer 8: 102.

    Article  Google Scholar 

  • Small EM, O'Rourke JR, Moresi V, Sutherland LB, McAnally J, Gerard RD et al. (2010). Regulation of PI3-kinase/Akt signaling by muscle-enriched microRNA-486. Proc Natl Acad Sci U S A 107: 4218–4223.

    Article  CAS  Google Scholar 

  • Theisen H, Syed A, Nguyen BT, Lukacsovich T, Purcell J, Srivastava GP et al. (2007). Wingless directly represses DPP morphogen expression via an armadillo/TCF/Brinker complex. PLoS One 2: e142.

    Article  Google Scholar 

  • Thorsen K, Sorensen KD, Brems-Eskildsen AS, Modin C, Gaustadnes M, Hein AM et al. (2008). Alternative splicing in colon, bladder, and prostate cancer identified by exon array analysis. Mol Cell Proteomics 7: 1214–1224.

    Article  CAS  Google Scholar 

  • Tsuchiya K, Nakamura T, Okamoto R, Kanai T, Watanabe M . (2007). Reciprocal targeting of Hath1 and beta-catenin by Wnt glycogen synthase kinase 3beta in human colon cancer. Gastroenterology 132: 208–220.

    Article  CAS  Google Scholar 

  • van de Wetering M, Sancho E, Verweij C, de Lau W, Oving I, Hurlstone A et al. (2002). The beta-catenin/TCF-4 complex imposes a crypt progenitor phenotype on colorectal cancer cells. Cell 111: 241–250.

    Article  CAS  Google Scholar 

  • Van der Flier LG, Sabates-Bellver J, Oving I, Haegebarth A, De Palo M, Anti M et al. (2007). The Intestinal Wnt/TCF Signature. Gastroenterology 132: 628–632.

    Article  CAS  Google Scholar 

  • Vlad A, Rohrs S, Klein-Hitpass L, Muller O . (2008). The first five years of the Wnt targetome. Cell Signal 20: 795–802.

    Article  CAS  Google Scholar 

  • Wang X, Lam EK, Zhang J, Jin H, Sung JJ . (2009). MicroRNA-122a functions as a novel tumor suppressor downstream of adenomatous polyposis coli in gastrointestinal cancers. Biochem Biophys Res Commun 387: 376–380.

    Article  CAS  Google Scholar 

  • Waterman ML . (2004). Lymphoid enhancer factor/T cell factor expression in colorectal cancer. Cancer Metastasis Rev 23: 41–52.

    Article  CAS  Google Scholar 

  • Weitz J, Koch M, Debus J, Hohler T, Galle PR, Buchler MW . (2005). Colorectal cancer. Lancet 365: 153–165.

    Article  Google Scholar 

  • Wu F, Zhu S, Ding Y, Beck WT, Mo YY . (2009). MicroRNA-mediated regulation of Ubc9 expression in cancer cells. Clin Cancer Res 15: 1550–1557.

    Article  CAS  Google Scholar 

  • Yochum GS, Cleland R, Goodman RH . (2008). A genome-wide screen for beta-catenin binding sites identifies a downstream enhancer element that controls c-Myc gene expression. Mol Cell Biol 28: 7368–7379.

    Article  CAS  Google Scholar 

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Acknowledgements

We thank Misti Martinez, Hanne Steen, Inge-Lis Thorsen and Pamela Celis for technical assistance. TOPflash/FOPflash plasmids were kindly provided by Philippe Blache, and the CLINT1 luciferase reporter plasmid was generously provided by Jesper B Bramsen. This work was supported by the Desirée and Niels Yde's Foundation, the John and Birthe Meyer Foundation, the Lundbeck Foundation, the Danish Council for Independent Research Medical Sciences, the Danish Council for Strategic Research, the Danish Agency for Science, Technology and Innovation, and the Danish Ministry of the Interior and Health.

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  1. Department of Molecular Medicine (MOMA), Aarhus University Hospital, Aarhus N, Denmark

    T Schepeler, A Holm, I Nordentoft, P Lamy, L L Christensen, K Thorsen, T F Ørntoft & C L Andersen

  2. Department of Biochemistry, Jim Ayers Institute for Precancer Detection and Diagnosis, Vanderbilt University School of Medicine, Nashville, TN, USA

    P Halvey & D C Liebler

  3. Biotech Research and Innovation Centre (BRIC), University of Copenhagen, Copenhagen, Denmark

    E M Riising & K Helin

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  1. T Schepeler
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  2. A Holm
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Correspondence to C L Andersen.

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Schepeler, T., Holm, A., Halvey, P. et al. Attenuation of the beta-catenin/TCF4 complex in colorectal cancer cells induces several growth-suppressive microRNAs that target cancer promoting genes. Oncogene 31, 2750–2760 (2012). https://doi.org/10.1038/onc.2011.453

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