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Increased invasiveness of osteosarcoma mesenchymal stem cells induced by bone-morphogenetic protein-2

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Abstract

To evaluate the different traits of mesenchymal stem cell (MSC) isolated from osteosarcoma (OS) and normal bone marrow (BM) induced by bone-morphogenetic protein-2 (BMP-2). MSCs from implanted osteosarcoma or femur bone marrow were isolated and cultured. Differentiation potency was verified and phenotypes were evaluated by flow cytometry. Increased or decreased expressions of BMP-2 were delivered by adenovirus and lentivirus vector, respectively. Expressions of VEGF, EMMPRIN, and MMP-9 were examined. Cell cycle, apoptosis, invasiveness, and proliferation assays were performed between the transfected groups and controls. Increased BMP-2 induced over-expression of VEGF, EMMPRIN, and MMP-9 in OS- and BM-MSCs both intra- and extra-cellularly. Decreased BMP-2 expression induced inhibition of the factors. Increased BMP-2 also induced less population of cells at G1 phase, more apoptotic cells, more cells that invade through Transwell membrane, and faster proliferation in OSMSC compared to those in BMMSC. BMP-2 induced higher expression of tumorigenic factors, which could be responsible for promoting the proliferation and aggressiveness of OSMSC over BMMSC.

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References

  • Alborzinia H, Schmidt-Glenewinkel H, Ilkavets I, Breitkopf-Heinlein K, Cheng X, Hortschansky P, Dooley S, Wölfl S. Quantitative kinetic analysis of BMP2 uptake into cells and its modulation by BMP-antagonists. J Cell Sci. 2012. doi:10.1242/jcs.109777.

  • Amado L. C.; Saliaris A. P.; Schuleri K. H.; St John M.; Xie J. S.; Cattaneo S.; Durand D. J.; Fitton T.; Kuang J. Q.; Stewart G.; Lehrke S.; Baumgartner W. W.; Martin B. J.; Heldman A. W.; Hare J. M. Cardiac repair with intramyocardial injection of allogeneic mesenchymal stem cells after myocardial infarction. Proc Natl Acad Sci U S A. 102: 11474–11479; 2005.

    Article  PubMed  CAS  Google Scholar 

  • Bergers G.; Brekken R.; McMahon G.; Vu T. H.; Itoh T.; Tamaki K.; Tanzawa K.; Thorpe P.; Itohara S.; Werb Z.; Hanahan D. Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. Nat Cell Biol. 2: 737–744; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Cheng H.; Fukushima T.; Takahashi N.; Tanaka H.; Kataoka H. Hepatocyte growth factor activator inhibitor type 1 regulates epithelial to mesenchymal transition through membrane-bound serine proteinases. Cancer Res 69: 1828–1835; 2009.

    Article  PubMed  CAS  Google Scholar 

  • Digirolamo C. M.; Stokes D.; Colter D.; Phinney D. G.; Class R.; Prockop D. J. Propagation and senescence of human marrow stromal cells in culture: a simple colony-forming assay identifies samples with the greatest potential to propagate and differentiate. Br J Haematol. 107: 275–281; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Dvorak H. F.; Brown L. F.; Detmar M.; Dvorak A. M. Vascular permeability factor/vascular endothelial growth factor, microvascular hyperpermeability, and angiogenesis. Am J Pathol. 146: 1029–1039; 1995.

    PubMed  CAS  Google Scholar 

  • Ghosh-Choudhury N.; Ghosh-Choudhury G.; Celeste A.; Ghosh P. M.; Moyer M.; Abboud S. L.; Kreisberg J. Bone morphogenetic protein-2 induces cyclin kinase inhibitor p21 and hypophosphorylation of retinoblastoma protein in estradiol-treated MCF-7 human breast cancer cells. Biochim Biophys Acta. 1497: 186–196; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Gouze J. N.; Stoddart M. J.; Gouze E.; Palmer G. D.; Ghivizzani S. C.; Grodzinsky A. J.; Evans C. H. In vitro gene transfer to chondrocytes and synovial fibroblasts by adenoviral vectors. Methods Mol Med. 100: 147–164; 2004.

    PubMed  CAS  Google Scholar 

  • Guo H.; Majmudar G.; Jensen T. C.; Biswas C.; Toole B. P.; Gordon M. K. Characterization of the gene for human EMMPRIN, a tumor cell surface inducer of matrix metalloproteinases. Gene. 220: 99–108; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Han J.; Yong B.; Luo C.; Tan P.; Peng T.; Shen J. High serum alkaline phosphatase cooperating with MMP-9 predicts metastasis and poor prognosis in patients with primary osteosarcoma in Southern China. World J Surg Oncol. 10: 37; 2012. doi:10.1186/1477-7819-10-37.

    Article  PubMed  Google Scholar 

  • Hardy S.; Kitamura M.; Harris-Stansil T.; Dai Y.; Phipps M. L. Construction of adenovirus vectors through Cre-lox recombination. J Virol. 71: 1842–1849; 1997.

    PubMed  CAS  Google Scholar 

  • Hatakeyama S.; Gao Y. H.; Ohara-Nemoto Y.; Kataoka H.; Satoh M. Expression of bone morphogenetic proteins of human neoplastic epithelial cells. Biochem Mol Biol Int. 42: 497–505; 1997.

    PubMed  CAS  Google Scholar 

  • Hicklin D. J.; Ellis L. M. Role of the vascular endothelial growth factor pathway in tumor growth and angiogenesis. J Clin Oncol 23: 1011–1027; 2005. Review.

    Article  PubMed  CAS  Google Scholar 

  • Igarashi M.; Yogiashi Y.; Mihara M.; Takada I.; Kitagawa H.; Kato S. Vitamin K induces osteoblast differentiation through pregnane X receptor-mediated transcriptional control of the Msx2 gene. Mol Cell Biol. 27: 7947–7954; 2007.

    Article  PubMed  CAS  Google Scholar 

  • Ji J. F.; He B. P.; Dheen S. T.; Tay S. S. Interactions of chemokines and chemokine receptors mediate the migration of mesenchymal stem cells to the impaired site in the brain after hypoglossal nerve injury. Stem Cells. 22: 415–427; 2004.

    Article  PubMed  CAS  Google Scholar 

  • Kaya M.; Wada T.; Kawaguchi S.; Nagoya S.; Yamashita T.; Abe Y.; Hiraga H.; Isu K.; Shindoh M.; Higashino F.; Okada F.; Tada M.; Yamawaki S.; Ishii S. Increased pre-therapeutic serum vascular endothelial growth factor in patients with early clinical relapse of osteosarcoma. Br J Cancer. 86: 864–869; 2002a.

    Article  PubMed  CAS  Google Scholar 

  • Kaya M.; Wada T.; Kawaguchi S.; Nagoya S.; Yamashita T.; Abe Y.; Hiraga H.; Isu K.; Shindoh M.; Higashino F.; Okada F.; Tada M.; Yamawaki S.; Ishii S. Increased pre-therapeutic serum vascular endothelial growth factor in patients with early clinical relapse of osteosarcoma. Br J Cancer. 86: 864–869; 2002b.

    Article  PubMed  CAS  Google Scholar 

  • Koç O. N.; Lazarus H. M. Mesenchymal stem cells: heading into the clinic. Bone Marrow Transplant 27: 235–239; 2001. Review.

    Article  PubMed  Google Scholar 

  • Li Y.; Xu J.; Chen L.; Zhong W. D.; Zhang Z.; Mi L.; Zhang Y.; Liao C. G.; Bian H. J.; Jiang J. L.; Yang X. M.; Li X. Y.; Fan C. M.; Zhu P.; Fu L.; Chen Z. N. HAb18G (CD147), a cancer-associated biomarker and its role in cancer detection. Histopathology 54: 677–687; 2009.

    Article  PubMed  CAS  Google Scholar 

  • Liotta L. A. Tumor invasion and metastases—role of the extracellular matrix: Rhoads Memorial Award lecture. Cancer Res 46: 1–7; 1986. Review.

    Article  PubMed  CAS  Google Scholar 

  • Liu Z.; Li Y.; Yang R. Effects of resveratrol on vascular endothelial growth factor expression in osteosarcoma cells and cell proliferation. Oncol Lett. 4(4): 837–839; 2012.

    PubMed  CAS  Google Scholar 

  • Massague J.; Chen Y. G. Controlling TGF-beta signaling. Genes Dev. 14: 627–644; 2000.

    PubMed  CAS  Google Scholar 

  • Meirelles Lda S.; Nardi N. B. Murine marrow-derived mesenchymal stem cell: isolation, in vitro expansion, and characterization. Br J Haematol. 123: 702–711; 2003.

    Article  PubMed  Google Scholar 

  • Muramatsu T.; Miyauchi T. Basigin (CD147): a multifunctional transmembrane protein involved in reproduction, neural function, inflammation and tumor invasion. Histol Histopathol 18: 981–987; 2003. Review.

    PubMed  CAS  Google Scholar 

  • Nishimura R.; Kato Y.; Chen D.; Harris S. E.; Mundy G. R.; Yoneda T. Smad5 and DPC4 are key molecules in mediating BMP-2-induced osteoblastic differentiation of the pluripotent mesenchymal precursor cell line C2C12. J Biol Chem. 273: 1872–1879; 1998.

    Article  PubMed  CAS  Google Scholar 

  • Nohe A.; Hassel S.; Ehrlich M.; Neubauer F.; Sebald W.; Henis Y. I.; Knaus P. The mode of bone morphogenetic protein (BMP) receptor oligomerization determines different BMP-2 signaling pathways. J Biol Chem. 277: 5330–5338; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Peister A.; Mellad J. A.; Larson B. L.; Hall B. M.; Gibson L. F.; Prockop D. J. Adult stem cells from bone marrow (MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential. Blood. 103: 1662–1668; 2004.

    Article  PubMed  CAS  Google Scholar 

  • Picci P.; Mercuri M.; Ferrari S.; Alberghini M.; Briccoli A.; Ferrari C.; Pignotti E.; Bacci G. Survival in high-grade osteosarcoma: improvement over 21 years at a single institution. Ann Oncol. 21: 1366–1373; 2010.

    Article  PubMed  CAS  Google Scholar 

  • Phinney D. G.; Kopen G.; Isaacson R. L.; Prockop D. J. Plastic adherent stromal cells from the bone marrow of commonly used strains of inbred mice: variations in yield, growth, and differentiation. J Cell Biochem. 72: 570–585; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Rastogi S.; Kumar R.; Sankineani S. R.; Marimuthu K.; Rijal L.; Prakash S.; Jalan D.; Khan S. A.; Sharma M. C. Role of vascular endothelial growth factor as a tumour marker in osteosarcoma: a prospective study. Int Orthop. 36(11): 2315–21; 2012.

    Article  PubMed  Google Scholar 

  • Rodriguez-Manzaneque J. C.; Lane T. F.; Ortega M. A.; Hynes R. O.; Lawler J.; Iruela-Arispe M. L. Thrombospondin-1 suppresses spontaneous tumor growth and inhibits activation of matrix metalloproteinase-9 and mobilization of vascular endothelial growth factor. Proc Natl Acad Sci U S A. 98: 12485–12490; 2001.

    Article  PubMed  CAS  Google Scholar 

  • Satake K.; Lou J.; Lenke L. G. Migration of mesenchymal stem cells through cerebrospinal fluid into injured spinal cord tissue. Spine (Phila Pa 1976) 29: 1971–1979; 2004.

    Article  Google Scholar 

  • Studeny M.; Marini F. C.; Dembinski J. L.; Zompetta C.; Cabreira-Hansen M.; Bekele B. N.; Champlin R. E.; Andreeff M. Mesenchymal stem cells: potential precursors for tumor stroma and targeted-delivery vehicles for anticancer agents. J Natl Cancer Inst. 96: 1593–1603; 2004.

    Article  PubMed  CAS  Google Scholar 

  • Takahashi Y.; Tucker S. L.; Kitadai Y.; Koura A. N.; Bucana C. D.; Cleary K. R.; Ellis L. M. Vessel counts and expression of vascular endothelial growth factor as prognostic factors in node-negative colon cancer. Arch Surg. 132: 541–546; 1997.

    Article  PubMed  CAS  Google Scholar 

  • Tuschl T.; Zamore P. D.; Lehmann R.; Bartel D. P.; Sharp P. A. Targeted mRNA degradation by double-stranded RNA in vitro. Genes Dev. 13: 3191–7; 1999.

    Article  PubMed  CAS  Google Scholar 

  • Vogel G. Embryonic stem cells. Stem cells not so stealthy after all. Science. 297: 175–177; 2002.

    Article  PubMed  CAS  Google Scholar 

  • Wynn R. F.; Hart C. A.; Corradi-Perini C.; O'Neill L.; Evans C. A.; Wraith J. E.; Fairbairn L. J.; Bellantuono I. A small proportion of mesenchymal stem cells strongly expresses functionally active CXCR4 receptor capable of promoting migration to bone marrow. Blood. 104: 2643–2645; 2004.

    Article  PubMed  CAS  Google Scholar 

  • Yan L.; Zucker S.; Toole B. P. Roles of the multifunctional glycoprotein, emmprin (basigin; CD147), in tumour progression. Thromb Haemost 93: 199–204; 2005.

    PubMed  CAS  Google Scholar 

  • Zhang G.; Li M.; Jin J.; Bai Y.; Yang C. Knockdown of S100A4 decreases tumorigenesis and metastasis in osteosarcoma cells by repression of matrix metalloproteinase-9. Asian Pac J Cancer Prev. 12(8): 2075–80; 2011.

    PubMed  Google Scholar 

  • Zhou Q.; Zhu Y.; Deng Z.; Long H.; Zhang S.; Chen X. VEGF and EMMPRIN expression correlates with survival of patients with osteosarcoma. Surg Oncol 20: 13–19; 2011. Review.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The authors report no conflict of interest. This study was supported in part by the Wuxi Health Bureau, no. YGZX1106.

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Authors and Affiliations

  1. Division of Orthopedics, Second People’s hospital of Wuxi, Nanjing Medical University, Wuxi, 214000, People’s Republic of China

    Xiao Yang, Ya-ping Wang, Ke Zeng, Ming-quan Qian, Gang Chen, Lei Shi & Guo-xing Zhu

  2. Division of Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University, Shanghai, China

    Feng-xiang Liu

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  1. Xiao Yang
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Correspondence to Guo-xing Zhu.

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Editor: T. Okamoto

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Yang, X., Wang, Yp., Liu, Fx. et al. Increased invasiveness of osteosarcoma mesenchymal stem cells induced by bone-morphogenetic protein-2. In Vitro Cell.Dev.Biol.-Animal 49, 270–278 (2013). https://doi.org/10.1007/s11626-013-9598-0

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  • DOI: https://doi.org/10.1007/s11626-013-9598-0

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