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BMP-6 inhibits MMP-9 expression by regulating heme oxygenase-1 in MCF-7 breast cancer cells

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Abstract

Purpose

BMP-6, which belongs to the TGF-β superfamily, is a multifunctional molecule with distinct abilities in embryogenesis and organogenesis. Our recent research has implied that BMP-6 may suppress breast cancer metastasis. In the present study, we extended to elucidate the molecular mechanism by which BMP-6 exerts its anti-tumorigenic effect.

Methods

The Boyden chamber assay was used to examine the ability of BMP-6 and HO-1 in MCF-7 malignant progress. RT-PCR, western blot, luciferase assay, and quantitative CHIP were used to determine the potential mechanism and signaling pathways by which BMP-6 and HO-1 function as anti-metastatic factors in MCF-7 cells.

Results

The Boyden chamber assay showed that BMP-6 inhibited the migration and invasion of MCF-7 cells, which effect was significantly deprived by knockdown of HO-1. We further demonstrated that BMP-6 treatment resulted in an activation of HO-1 transcription through the recruitment of Smad1/5 to the Smad-responsive element on its promoter. In addition, BMP-6-induced up-regulation of HO-1 exhibited an inhibitory effect on MMP-9 secretion in a paracrine action in MCF-7 cells. Overexpression of BMP-6 and HO-1 synergistically suppressed MMP-9 transcription, which effect was specifically mediated via the MAPK/p38/AP-1 signaling. However, blockade of HO-1 using ZnPPIX totally abolished BMP-6-regulated MMP-9 activation in MCF-7 cells.

Conclusions

These observations suggest a novel role of BMP-6/HO-1 cascade to relieve breast cancer metastasis by regulating the secretion of growth factors in tumor microenvironment.

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References

  • Alarmo EL, Rauta J, Kauraniemi P, Karhu R, Kuukasjärvi T, Kallioniemi A (2006) Bone morphogenetic protein 7 is widely overexpressed in primary breast cancer. Genes Chromosomes Cancer 45:411–419

    Article  PubMed  CAS  Google Scholar 

  • Alarmo EL, Pärssinen J, Ketolainen JM, Savinainen K, Karhu R, Kallioniemi A (2009) BMP7 influences proliferation, migration, and invasion of breast cancer cells. Cancer Lett 275:35–43

    Article  PubMed  CAS  Google Scholar 

  • Becker JC, Fukui H, Imai Y, Sekikawa A, Kimura T, Yamagishi H, Yoshitake N, Pohle T, Domschke W, Fujimori T (2007) Colonic expression of heme oxygenase-1 is associated with a better long-term survival in patients with colorectal cancer. Scand J Gastroenterol 42:852–858

    Article  PubMed  CAS  Google Scholar 

  • Bendrik C, Robertson J, Gauldie J, Dabrosin C (2008) Gene transfer of matrix metalloproteinase-9 induces tumor regression of breast cancer in vivo. Cancer Res 68:3405–3412

    Article  PubMed  CAS  Google Scholar 

  • Blick T, Widodo E, Hugo H, Waltham M, Lenburg ME, Neve RM, Thompson EW (2008) Epithelial mesenchymal transition traits in human breast cancer cell lines. Clin Exp Metastasis 25:629–642

    Article  PubMed  CAS  Google Scholar 

  • Caballero F, Meiss R, Gimenez A, Batlle A, Vazquez E (2004) Immunohistochemical analysis of heme oxygenase-1 in preneoplastic and neoplastic lesions during chemical hepatocarcinogenesis. Int J Exp Pathol 85:213–222

    Article  PubMed  CAS  Google Scholar 

  • Celeste AJ, Iannazzi JA, Taylo RC, Hewick RM, Rosen V, Wang EA, Wozney JM (1990) Identification of transforming growth factor beta family members present in bone-inductive protein purified from bovine bone. Proc Natl Acad Sci USA 87:9843–9847

    Article  PubMed  CAS  Google Scholar 

  • Chabottaux V, Noel A (2007) Breast cancer progression: insights into multifaceted matrix metalloproteinases. Clin Exp Metastasis 24:647–656

    Article  PubMed  CAS  Google Scholar 

  • Clement JH, Sanger J, Hoffken K (1999) Expression of bone morphogenetic protein 6 in normal mammary tissue and breast cancer cell lines and its regulation by epidermal growth factor. Int J Cancer 80:250–256

    Article  PubMed  CAS  Google Scholar 

  • Cornblatt BS, Ye L, Dinkova-Kostova AT, Erb M, Fahey JW, Singh NK, Chen MS, Stierer T, Garrett-Mayer E, Argani P, Davidson NE, Talalay P, Kensler TW, Visvanathan K (2007) Preclinical and clinical evaluation of sulforaphane for chemoprevention in the breast. Carcinogenesis 28:1485–1490

    Article  PubMed  CAS  Google Scholar 

  • Derynck R, Zhang YE (2003) Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature 425:577–584

    Article  PubMed  CAS  Google Scholar 

  • Du J, Yang S, An D, Hu F, Yuan W, Zhai C, Zhu T (2009) BMP-6 inhibits microRNA-21 expression in breast cancer through repressing deltaEF1 and AP-1. Cell Res 19:487–496

    Article  PubMed  CAS  Google Scholar 

  • Ducy P, Karsenty G (2000) The family of bone morphogenetic proteins. Kidney Int 57:2207–2214

    Article  PubMed  CAS  Google Scholar 

  • Eferl R, Wagner EF (2003) AP-1: a double-edged sword in tumorigenesis. Nat Rev Cancer 3:859–868

    Article  PubMed  CAS  Google Scholar 

  • Egeblad M, Werb Z (2002) New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2:161–174

    Article  PubMed  CAS  Google Scholar 

  • Ferenbach DA, Kluth DC, Hughes J (2010) Hemeoxygenase-1 and renal ischaemia-reperfusion injury. Nephron Exp Nephrol 115:e33–e37

    Article  PubMed  CAS  Google Scholar 

  • Gitelman SE, Kobrin MS, Ye JQ, Lopez AR, Lee A, Derynck R (1994) Recombinant Vgr-1/BMP-6-expressing tumors induce fibrosis and enchondral bone formation in vivo. J Cell Biol 126:1595–1609

    Article  PubMed  CAS  Google Scholar 

  • Gitelman SE, Kirk M, Ye JQ, Filvaroff EH, Kahn AJ, Derynck R (1995) Vgr-1/BMP-6 induces osteoblast differentiation of pluripotential mesenchymal cells. Cell Growth Differ 67:827–836

    Google Scholar 

  • Hill M, Pereira V, Chauveau C, Zagani R, Remy S, Tesson L, Mazal D, Ubillos L, Brion R, Asghar K, Mashreghi MF, Kotsch K, Moffett J, Doebis C, Seifert M, Boczkowski J, Osinaga E, Anegon I (2005) Heme oxygenase-1 inhibits rat and human breast cancer cell proliferation: mutual cross inhibition with indoleamine 2, 3-dioxygenase. FASEB J 19:1957–1968

    Article  PubMed  CAS  Google Scholar 

  • Hill-Kapturczak N, Sikorski E, Voakes C, Garcia J, Nick HS, Agarwal A (2003) An internal enhancer regulates heme- and cadmium-mediated induction of human heme oxygenase-1. Am J Physiol Renal Physiol 285:F515–F523

    PubMed  Google Scholar 

  • Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ (2009) Cancer statistics. CA Cancer J Clin 59:225–249

    Google Scholar 

  • Jespersen C, Doller A, Akool el S, Bachmann M, Müller R, Gutwein P, Mühl H, Pfeilschifter J, Eberhardt W (2009) Molecular mechanisms of nitric oxide-dependent inhibition of TPA-induced matrix metalloproteinase-9 (MMP-9) in MCF-7 cells. J Cell Physiol 219:276–287

    Article  PubMed  CAS  Google Scholar 

  • Kang Y, Massagué J (2004) Epithelial-mesenchymal transitions: twist in development and metastasis. Cell 118:277–279

    Article  PubMed  CAS  Google Scholar 

  • Keum YS, Yu S, Chang PP, Yuan X, Kim JH, Xu C, Han J, Agarwal A, Kong AN (2006) Mechanism of action of sulforaphane: inhibition of p38 mitogen-activated protein kinase isoforms contributing to the induction of antioxidant response element-mediated heme oxygenase-1 in human hepatoma HepG2 cells. Cancer Res 66:8804–8813

    Article  PubMed  CAS  Google Scholar 

  • Kie JH, Kapturczak MH, Traylor A, Agarwal A, Hill-Kapturczak N (2008) Heme oxygenase-1 deficiency promotes epithelial-mesenchymal transition and renal fibrosis. J Am Soc Nephrol 19:1681–1691

    Article  PubMed  CAS  Google Scholar 

  • Kim S, Choi JH, Kim JB, Nam SJ, Yang JH, Kim JH, Lee JE (2008) Berberine suppresses TNF-alpha-induced MMP-9 and cell invasion through inhibition of AP-1 activity in MDA-MB-231 human breast cancer cells. Molecules 13:2975–2985

    Article  PubMed  CAS  Google Scholar 

  • Kumagai T, Shimizu T, Takeda K (2006) Bone morphogenetic protein-2 suppresses invasiveness of TSU-Pr1 cells with the inhibition of MMP-9 secretion. Anticancer Res 26:293–298

    PubMed  CAS  Google Scholar 

  • Lee GT, Kwon SJ, Lee JH, Jeon SS, Jang KT, Choi HY, Lee HM, Kim WJ, Kim SJ, Kim IY (2010) Induction of interleukin-6 expression by bone morphogenetic protein-6 in macrophages requires both Smad and P38 signaling pathways. J Biol Chem [Epub ahead of print]

  • Lin CW, Shen SC, Hou WC, Yang LY, Chen YC (2008) Heme oxygenase-1 inhibits breast cancer invasion via suppressing the expression of matrix metalloproteinase-9. Mol Cancer Ther 7:1195–1206

    Article  PubMed  CAS  Google Scholar 

  • Mi D, Zhang M, Yan JD, Zhang J, Wang X, Wang Q, Yang S, Zhu TH (2010) PTHrP inhibits BMP-6 expression through the PKA signaling pathway in breast cancer cells. J Cancer Res Clin Oncol [Epub ahead of print]

  • Morse D, Lin L, Choi AM, Ryter SW (2009) Heme oxygenase-1, a critical arbitrator of cell death pathways in lung injury and disease. Free Radic Biol Med 47:1–12

    Article  PubMed  CAS  Google Scholar 

  • Nagaiah G, Abraham J (2010) Circulating tumor cells in the management of breast cancer. Clin Breast Cancer 10:209–216

    Article  PubMed  CAS  Google Scholar 

  • Pellikainen JM, Ropponen KM, Kataja VV, Kellokoski JK, Eskelinen MJ, Kosma VM (2004) Expression of matrix metalloproteinase (MMP)-2 and MMP-9 in breast cancer with a special reference to activator protein-2, HER2, and prognosis. Clin Cancer Res 10:7621–7628

    Article  PubMed  CAS  Google Scholar 

  • Reinholz MM, Iturria SJ, Ingle JN, Roche PC (2002) Differential gene expression of TGF-beta family members and osteopontin in breast tumor tissue: analysis by real-time quantitative PCR. Breast Cancer Res Treat 74:255–269

    Article  PubMed  CAS  Google Scholar 

  • Ryter SW, Alam J, Choi AM (2006) Heme oxygenase-1/carbon monoxide: from basic science to therapeutic applications. Physiol Rev 86:583–650

    Article  PubMed  CAS  Google Scholar 

  • Safina A, Ren MQ, Vandette E, Bakin AV (2008) TAK1 is required for TGF-beta 1-mediated regulation of matrix metalloproteinase-9 and metastasis. Oncogene 27:1198–1207

    Article  PubMed  CAS  Google Scholar 

  • Shon SK, Kim A, Kim JY, Kim KI, Yang Y, Lim JS (2009) Bone morphogenetic protein-4 induced by NDRG2 expression inhibits MMP-9 activity in breast cancer cells. Biochem Biophys Res Commun 385:198–203

    Article  PubMed  CAS  Google Scholar 

  • Stallings-Mann M, Radisky D (2007) Matrix metalloproteinase-induced malignancy in mammary epithelial cells. Cells Tissues Organs 185:104–110

    Article  PubMed  CAS  Google Scholar 

  • Takahashi M, Otsuka F, Miyoshi T, Otani H, Goto J, Yamashita M, Ogura T, Makino H, Doihara H (2008) Bone morphogenetic protein 6 (BMP6) and BMP7 inhibit estrogen-induced proliferation of breast cancer cells by suppressing p38 mitogen-activated protein kinase activation. J Endocrinol 199:445–455

    Article  PubMed  CAS  Google Scholar 

  • Tang SJ, Hoodless PA, Lu Z, Breitman ML, McInnes RR, Wrana JL, Buchwald M (1998) The Tlx-2 homeobox gene is a downstream target of BMP signalling and is required for mouse mesoderm development. Development 125:1877–1887

    PubMed  CAS  Google Scholar 

  • Tsuji MH, Yanagawa T, Iwasa S, Tabuchi K, Onizawa K, Bannai S, Toyooka H, Yoshida H (1999) Heme oxygenase-1 expression in oral squamous cell carcinoma as involved in lymph node metastasis. Cancer Lett 138:53–59

    Article  PubMed  CAS  Google Scholar 

  • Vukicevic S, Grgurevic L (2009) BMP-6 and mesenchymal stem cell differentiation. Cytokine Growth Factor Rev 20:441–448

    Article  PubMed  CAS  Google Scholar 

  • Wozney JM (1992) The bone morphogenetic protein family and osteogenesis. Mol Reprod Dev 32:160–167

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto Y, Oelgeschläger M (2004) Regulation of bone morphogenetic proteins in early embryonic development. Naturwissenschaften 91:519–534

    Article  PubMed  CAS  Google Scholar 

  • Yan JD, Yang S, Lü SJ, Lei RY, Zhu TH (2007) Expression of recombinant human BMP6 in CHO cells by fused to the signal peptide and propeptide of another homologue protein. Sheng Wu Gong Cheng Xue Bao 23:413–417

    PubMed  CAS  Google Scholar 

  • Yan J, Yang S, Zhang J, Zhai C, Zhu T (2009) BMP6 attenuates oxidant injury in HK-2 cells via Smad-dependent HO-1 induction. Free Radic Biol Med 46:1275–1282

    Article  PubMed  CAS  Google Scholar 

  • Yang S, Du J, Wang Z, Yuan W, Qiao Y, Zhang M, Zhang J, Gao S, Yin J, Sun B, Zhu TH (2007) BMP-6 promotes E-cadherin expression through repressing deltaEF1 in breast cancer cells. BMC Cancer 7:211

    Article  PubMed  Google Scholar 

  • Yang S, Du J, Wang Z, Yan J, Yuan W, Zhang J, Zhu T (2009) Dual mechanism of deltaEF1 expression regulated by bone morphogenetic protein-6 in breast cancer. Int J Biochem Cell Biol 41:853–861

    Article  PubMed  CAS  Google Scholar 

  • Zhang M, Wang Q, Yuan W, Yang S, Wang X, Yan JD, Du J, Yin J, Gao SY, Sun BC, Zhu TH (2007) Epigenetic regulation of bone morphogenetic protein-6 gene expression in breast cancer cells. J Steroid Biochem Mol Biol 105:91–97

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work is supported by grants from the National Nature Science Foundation of China (No. 81072153) and from Tianjin Natural Science Foundation (No. 09JCYBJC09100) to S. Yang.

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The authors declare that we have no conflict of interest.

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Correspondence to Shuang Yang.

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Wang, C., Hu, F., Guo, S. et al. BMP-6 inhibits MMP-9 expression by regulating heme oxygenase-1 in MCF-7 breast cancer cells. J Cancer Res Clin Oncol 137, 985–995 (2011). https://doi.org/10.1007/s00432-010-0963-z

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  • DOI: https://doi.org/10.1007/s00432-010-0963-z

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