Tumor Biology

, Volume 36, Issue 12, pp 9475–9486 | Cite as

BMP-2 induces motility and invasiveness by promoting colon cancer stemness through STAT3 activation

  • Bo Ram Kim
  • Sang Cheul Oh
  • Dae-Hee Lee
  • Jung Lim Kim
  • Suk Young Lee
  • Myoung Hee Kang
  • Sun Il Lee
  • Sanghee Kang
  • Sung Yup Joung
  • Byung Wook Min
Research Article

Abstract

Bone morphogenetic proteins (BMPs) have been involved in metastatic progression and tumorigenesis of many cancer types. However, it remains unclear how BMP-2 contributes to the initiation and development of these cancers. Here, we investigated the role of BMP-2 in colon cancer stem cell (CSC) development from colon cancer cells. We also determined the effects of BMP-2 on CSC development and epithelial-mesenchymal transition (EMT) in human colon cancer cell lines HCT-116 and SW620. We found that BMP-2 enhanced sphere formation of colon cancer cells without serum. Also, BMP-2-induced spheres displayed up-regulation of stemness markers (CD133+ and EpCAM+) and increased drug resistance, hallmarks of CSCs. Importantly, expression of EMT activators p-Smad1/5 and Snail and N-cadherin was increased in the spheres’ cells, indicating that BMP-2 signaling might result in CSC self-renewal and EMT. Furthermore, siRNA-mediated knockdown of signal transducer and activator of transcription 3 (STAT3) in HCT-116 cells reversed BMP-2-induced EMT and stem cell formation. Taken together, our results suggest that the BMP-2 induced STAT3-mediated induction of colon cancer cell metastasis requires an EMT and/or changes in CSC markers.

Keywords

Colon cancer Bone morphogenetic proteins Cancer stem cells Epithelial-mesenchymal transition Signal transducer and activator of transcription 3 

Notes

Acknowledgments

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education (NRF2012R1A1A2042905) and supported by Korea University Grant (K1421591).

Conflicts of interest

None

References

  1. 1.
    Wolpin BM, Mayer RJ. Systemic treatment of colorectal cancer. Gastroenterology. 2008;134:1296–310.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Lai YL, Lin JK, Liang WY, Huang YC, Chang SC. Surgical resection combined with chemotherapy can help achieve better outcomes in patients with primary colonic lymphoma. J Surg Oncol. 2011;104:265–8.CrossRefPubMedGoogle Scholar
  3. 3.
    Lobo NA, Shimono Y, Qian D, Clarke MF. The biology of cancer stem cells. Annu Rev Cell Dev Biol. 2007;23:675–99.CrossRefPubMedGoogle Scholar
  4. 4.
    Smalley MJ, Dale TC. Wnt signalling in mammalian development and cancer. Cancer Metastasis Rev. 1999;18:215–30.CrossRefPubMedGoogle Scholar
  5. 5.
    Liu S, Dontu G, Mantle ID, Patel S, Ahn NS, Jackson KW, et al. Hedgehog signaling and bmi-1 regulate self-renewal of normal and malignant human mammary stem cells. Cancer Res. 2006;66:6063–71.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Dontu G, Jackson KW, McNicholas E, Kawamura MJ, Abdallah WM, Wicha MS. Role of notch signaling in cell-fate determination of human mammary stem/progenitor cells. Breast Cancer Res: BCR. 2004;6:R605–15.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Yeung TM, Gandhi SC, Wilding JL, Muschel R, Bodmer WF. Cancer stem cells from colorectal cancer-derived cell lines. Proc Natl Acad Sci U S A. 2010;107:3722–7.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Dalerba P, Dylla SJ, Park IK, Liu R, Wang X, Cho RW, et al. Phenotypic characterization of human colorectal cancer stem cells. Proc Natl Acad Sci U S A. 2007;104:10158–63.CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Hardwick JCH, Van Den Brink GR, Bleuming SA, Ballester I, Van Den Brande JMH, Keller JJ, et al. Bone morphogenetic protein 2 is expressed by, and acts upon, mature epithelial cells in the colon. Gastroenterology. 2004;126:111–21.CrossRefPubMedGoogle Scholar
  10. 10.
    Miyazono K, Kusanagi K, Inoue H. Divergence and convergence of tgf-beta/bmp signaling. J Cell Physiol. 2001;187:265–76.CrossRefPubMedGoogle Scholar
  11. 11.
    Miyazawa K, Shinozaki M, Hara T, Furuya T, Miyazono K. Two major smad pathways in tgf-beta superfamily signalling. Genes Cells: Devoted Mol Cell Mech. 2002;7:1191–204.CrossRefGoogle Scholar
  12. 12.
    Shirai YT, Ehata S, Yashiro M, Yanagihara K, Hirakawa K, Miyazono K. Bone morphogenetic protein-2 and -4 play tumor suppressive roles in human diffuse-type gastric carcinoma. Am J Pathol. 2011;179:2920–30.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Zhang Y, Chen X, Qiao M, Zhang BQ, Wang N, Zhang Z, et al. Bone morphogenetic protein 2 inhibits the proliferation and growth of human colorectal cancer cells. Oncol Rep. 2014;32:1013–20.PubMedPubMedCentralGoogle Scholar
  14. 14.
    Kang MH, Oh SC, Lee HJ, Kang HN, Kim JL, Kim JS, et al. Metastatic function of bmp-2 in gastric cancer cells: the role of pi3k/akt, mapk, the nf-kappab pathway, and mmp-9 expression. Exp Cell Res. 2011;317:1746–62.CrossRefPubMedGoogle Scholar
  15. 15.
    Fong YC, Li TM, Wu CM, Hsu SF, Kao ST, Chen RJ, et al. Bmp-2 increases migration of human chondrosarcoma cells via pi3k/akt pathway. J Cell Physiol. 2008;217:846–55.CrossRefPubMedGoogle Scholar
  16. 16.
    Langenfeld EM, Calvano SE, Abou-Nukta F, Lowry SF, Amenta P, Langenfeld J. The mature bone morphogenetic protein-2 is aberrantly expressed in non-small cell lung carcinomas and stimulates tumor growth of a549 cells. Carcinogenesis. 2003;24:1445–54.CrossRefPubMedGoogle Scholar
  17. 17.
    Thiery JP. Epithelial–mesenchymal transitions in development and pathologies. Curr Opin Cell Biol. 2003;15:740–6.CrossRefPubMedGoogle Scholar
  18. 18.
    Bedi S, Vidyasagar A, Djamali A. Epithelial-to-mesenchymal transition and chronic allograft tubulointerstitial fibrosis. Transplant Rev. 2008;22:1–5.CrossRefGoogle Scholar
  19. 19.
    Moustakas A, Heldin CH. Signaling networks guiding epithelial-mesenchymal transitions during embryogenesis and cancer progression. Cancer Sci. 2007;98:1512–20.CrossRefPubMedGoogle Scholar
  20. 20.
    Tsai JH, Yang J. Epithelial-mesenchymal plasticity in carcinoma metastasis. Genes Dev. 2013;27:2192–206.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Thiery JP, Acloque H, Huang RY, Nieto MA. Epithelial-mesenchymal transitions in development and disease. Cell. 2009;139:871–90.CrossRefPubMedGoogle Scholar
  22. 22.
    Gupta PB, Chaffer CL, Weinberg RA. Cancer stem cells: mirage or reality? Nat Med. 2009;15:1010–2.CrossRefPubMedGoogle Scholar
  23. 23.
    Kang MH, Kim JS, Seo JE, Oh SC, Yoo YA. Bmp2 accelerates the motility and invasiveness of gastric cancer cells via activation of the phosphatidylinositol 3-kinase (pi3k)/akt pathway. Exp Cell Res. 2010;316:24–37.CrossRefPubMedGoogle Scholar
  24. 24.
    Kryczek I, Lin Y, Nagarsheth N, Peng D, Zhao L, Zhao E, et al. Il-22(+)cd4(+) t cells promote colorectal cancer stemness via stat3 transcription factor activation and induction of the methyltransferase dot1l. Immunity. 2014;40:772–84.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Lejeune D, Dumoutier L, Constantinescu S, Kruijer W, Schuringa JJ, Renauld JC. Interleukin-22 (il-22) activates the jak/stat, erk, jnk, and p38 map kinase pathways in a rat hepatoma cell line. Pathways that are shared with and distinct from il-10. J Biol Chem. 2002;277:33676–82.CrossRefPubMedGoogle Scholar
  26. 26.
    Pickert G, Neufert C, Leppkes M, Zheng Y, Wittkopf N, Warntjen M, et al. Stat3 links il-22 signaling in intestinal epithelial cells to mucosal wound healing. J Exp Med. 2009;206:1465–72.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Hogan BL. Bone morphogenetic proteins in development. Curr Opin Genet Dev. 1996;6:432–8.CrossRefPubMedGoogle Scholar
  28. 28.
    Urist MR. Bone: formation by autoinduction. Science (New York, NY). 1965;150:893–9.CrossRefGoogle Scholar
  29. 29.
    Reddi AH. Bone and cartilage differentiation. Curr Opin Genet Dev. 1994;4:737–44.CrossRefPubMedGoogle Scholar
  30. 30.
    Liao A, Wang W, Sun D, Jiang Y, Tian S, Li J, Yang X, Shi R. Bone morphogenetic protein 2 mediates epithelial-mesenchymal transition via akt and erk signaling pathways in gastric cancer. Tumour Biol. 2015;36:2773–78.Google Scholar
  31. 31.
    Hou CH, Hsiao YC, Fong YC, Tang CH. Bone morphogenetic protein-2 enhances the motility of chondrosarcoma cells via activation of matrix metalloproteinase-13. Bone. 2009;44:233–42.CrossRefPubMedGoogle Scholar
  32. 32.
    Kang MH, Kang HN, Kim JL, Kim JS, Oh SC, Yoo YA. Inhibition of pi3 kinase/akt pathway is required for bmp2-induced emt and invasion. Oncol Rep. 2009;22:525–34.PubMedGoogle Scholar
  33. 33.
    Piccirillo SG, Vescovi AL. Bone morphogenetic proteins regulate tumorigenicity in human glioblastoma stem cells. Ernst Schering Found Symp Proc. 2006;5:59–81.Google Scholar
  34. 34.
    Shmelkov SV, Butler JM, Hooper AT, Hormigo A, Kushner J, Milde T, et al. Cd133 expression is not restricted to stem cells, and both cd133+ and cd133- metastatic colon cancer cells initiate tumors. J Clin Investig. 2008;118:2111–20.PubMedPubMedCentralGoogle Scholar
  35. 35.
    Langan RC, Mullinax JE, Raiji MT, Upham T, Summers T, Stojadinovic A, et al. Colorectal cancer biomarkers and the potential role of cancer stem cells. J Cancer. 2013;4:241–50.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Chambers I, Tomlinson SR. The transcriptional foundation of pluripotency. Development. 2009;136:2311–22.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Liao T, Kaufmann AM, Qian X, Sangvatanakul V, Chen C, Kube T, et al. Susceptibility to cytotoxic t cell lysis of cancer stem cells derived from cervical and head and neck tumor cell lines. J Cancer Res Clin Oncol. 2013;139:159–70.CrossRefPubMedGoogle Scholar
  38. 38.
    Lu Y, Zhu H, Shan H, Lu J, Chang X, Li X, et al. Knockdown of oct4 and nanog expression inhibits the stemness of pancreatic cancer cells. Cancer Lett. 2013;340:113–23.CrossRefPubMedGoogle Scholar
  39. 39.
    Ma L, Lu MF, Schwartz RJ, Martin JF. Bmp2 is essential for cardiac cushion epithelial-mesenchymal transition and myocardial patterning. Development. 2005;132:5601–11.CrossRefPubMedGoogle Scholar
  40. 40.
    Ghosh-Choudhury N, Mandal CC, Choudhury GG. Statin-induced ras activation integrates the phosphatidylinositol 3-kinase signal to akt and mapk for bone morphogenetic protein-2 expression in osteoblast differentiation. J Biol Chem. 2007;282:4983–93.CrossRefPubMedGoogle Scholar
  41. 41.
    Ghosh-Choudhury N, Abboud SL, Nishimura R, Celeste A, Mahimainathan L, Choudhury GG. Requirement of bmp-2-induced phosphatidylinositol 3-kinase and akt serine/threonine kinase in osteoblast differentiation and smad-dependent bmp-2 gene transcription. J Biol Chem. 2002;277:33361–8.CrossRefPubMedGoogle Scholar
  42. 42.
    Kashyap V, Rezende NC, Scotland KB, Shaffer SM, Persson JL, Gudas LJ, et al. Regulation of stem cell pluripotency and differentiation involves a mutual regulatory circuit of the nanog, oct4, and sox2 pluripotency transcription factors with polycomb repressive complexes and stem cell micrornas. Stem Cells Dev. 2009;18:1093–108.CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Kim D, Kim CH, Moon JI, Chung YG, Chang MY, Han BS, et al. Generation of human induced pluripotent stem cells by direct delivery of reprogramming proteins. Cell Stem Cell. 2009;4:472–6.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Bo Ram Kim
    • 1
  • Sang Cheul Oh
    • 2
  • Dae-Hee Lee
    • 2
  • Jung Lim Kim
    • 2
  • Suk Young Lee
    • 2
  • Myoung Hee Kang
    • 3
  • Sun Il Lee
    • 4
  • Sanghee Kang
    • 4
  • Sung Yup Joung
    • 4
  • Byung Wook Min
    • 4
  1. 1.Graduate School of MedicineKorea University College of MedicineSeoulRepublic of Korea
  2. 2.Division of Oncology/Hematology, Department of Internal MedicineKorea University College of MedicineSeoulRepublic of Korea
  3. 3.University of Ulsan College of MedicineAsan Institute for Life ScienceSeoulRepublic of Korea
  4. 4.Department of Surgery, Korea University Guro HospitalKorea University College of MedicineSeoulRepublic of Korea

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