Current Medical Science

, Volume 38, Issue 5, pp 809–817 | Cite as

MMP-1 Over-expression Promotes Malignancy and Stem-Like Properties of Human Osteosarcoma MG-63 Cells In Vitro

  • Man-li Tang
  • Xiang-jun Bai
  • Yong Li
  • Xiao-jing Dai
  • Fan YangEmail author


Osteosarcoma is the most common primary malignant bone tumor in childhood, and it maintains a high level of recurrence. Matrix metalloproteinase-1 (MMP-1) was found to contribute to cancer progression. The present study was to investigate the in vitro effects of MMP-1 over-expression on the proliferation, invasion, metastasis and stem-like properties of osteosarcoma MG-63 cells. The MG-63 cells were cultured and had a full length MMP-1 cDNA inserted by the lentiviral vector (MG-63MMP-1+). MG-63 negative control and MG-63 blank control groups were established as well. MMP-1 expression was detected in MG-63MMP-1+, MG-63 negative control and MG-63 blank control cells using qPCR, Western blotting and immunofluorescence after 24 h of culture. The cell proliferation assay was performed with a camera attached to a bioreactor, which was programmed to photograph five regions of each well every 10 min over a period of 48 h. The cell invasion assay was conducted with Matrigel to assess the invasive potential of MG-63 cells over 24 h, the qPCR analysis to measure stem cell markers, including Oct4, Sox-2, Nanog, and Pax-7, and Western blot analysis to detect invasive and metastatic potential markers TIMP-1, VEGF and BMP2/4, after 24 h of culture. Immunofluorescence was used to investigate the presence of the stem cell marker Pax-7 after 24-h culture. The results showed that over-expression of MMP-1 after transfection could significantly increase tumor cell proliferation and invasion (P<0.05, MG-63MMP-1+versus controls). Pax-7 was highly expressed in MG-63MMP-1+ cells, with no significant changes of Oct-4, Sox-2, and Nanog observed (P<0.05). MG-63MMP-1+ cells showed higher expression of VEGF and BMP 2/4 proteins and lower expression of TIMP-1 protein than controls (P<0.05). It was concluded that MMP-1 over-expression in MG-63 cells contributed to the proliferation, invasion, metastasis and stem-like properties of osteosarcoma cells. Future studies should focus on in vivo effects of MMP-1 over-expression and the application of MMP-1 and Pax-7 inhibition in vivo to osteosarcoma therapies.

Key words

osteosarcoma MG-63 cells matrix metalloproteinase-1 proliferation invasion metastasis 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Kim HJ, Chalmers PN, Morris CD. Pediatric osteosarcoma. Curr Opin Pediatr, 2010,22(1):61–66CrossRefGoogle Scholar
  2. 2.
    Osasan S, Zhang M, Shen F, et al. Osteosarcoma: A 21st century review. Anticancer Res, 2016,36(9):4391–4398CrossRefGoogle Scholar
  3. 3.
    Friebele JC, Peck J, Pan X, et al. Osteosarcoma: A meta-Analysis and review of the literature. Am J Orthop, 2015,44(12):547–553Google Scholar
  4. 4.
    Kansara M, Teng MW, Smyth MJ, et al. Translational biology of osteosarcoma. Nat Rev Cancer, 2014,14(11):722–735CrossRefGoogle Scholar
  5. 5.
    Huang G, Nishimoto K, Yang Y, et al. Participation of the Fas/FasL signaling pathway and the lung microenvironment in the development of osteosarcoma lung metastases. Adv Exp Med Biol, 2014,804:203–217CrossRefGoogle Scholar
  6. 6.
    Li N, Yang R, Zhang W, et al. Genetically transforming human mesenchymal stem cells to sarcomas: changes in cellular phenotype and multilineage differentiation potential. Cancer, 2009,115(20):4795–4806CrossRefGoogle Scholar
  7. 7.
    Burns JS, Abdallah BM, Guldberg P, et al. Tumorigenic heterogeneity in cancer stem cells evolved from long-term cultures of telomerase-immortalized human mesenchymal stem cells. Cancer Res, 2005,65(8):3126–3135CrossRefGoogle Scholar
  8. 8.
    Ratajczak MZ, Zuba-Surma E, Kucia M, et al. Pluripotent and multipotent stem cells in adult tissues. Adv Med Sci, 2012,57(1):1–17CrossRefGoogle Scholar
  9. 9.
    Honoki K, Fujii H, Kubo A, et al. Possible involvement of stem-like populations with elevated ALDH1 in sarcomas for chemotherapeutic drug resistance. Oncol Rep, 2010,24(2):501–505CrossRefGoogle Scholar
  10. 10.
    Levings PP, McGarry SV, Currie TP, et al. Expression of an exogenous human Oct-4 promoter identifies tumor-initiating cells in osteosarcoma. Cancer Res, 2009,69(14):5648–5655CrossRefGoogle Scholar
  11. 11.
    Buckingham M, Relaix F. PAX3 and PAX7 as upstream regulators of myogenesis. Semin Cell Dev Biol, 2015,44:115–125CrossRefGoogle Scholar
  12. 12.
    Li CG, Eccles MR. PAX genes in cancer: Friends or foes? Front Genet, 2012, 31(3):6Google Scholar
  13. 13.
    Vargová V, Pytliak M, Mechírová V. Matrix metalloproteinases. EXS, 2012,103:1–33Google Scholar
  14. 14.
    Yadav L, Puri N, Rastogi V, et al. Matrix metalloproteinases and cancer-roles in threat and therapy. Asian Pac J Cancer Prev, 2014,15(3):1085–1091CrossRefGoogle Scholar
  15. 15.
    Pickup MW, Mouw JK, Weaver VM. The extracellular matrix modulates the hallmarks of cancer. EMBO Rep, 2014,15(12):1243–1253CrossRefGoogle Scholar
  16. 16.
    Hadler-Olsen E, Winberg JO, Uhlin-Hansen L. Matrix metalloproteinases in cancer: their value as diagnostic and prognostic markers and therapeutic targets. Tumour Bio, 2013,34(4):2041–2051CrossRefGoogle Scholar
  17. 17.
    Pulukuri SM, Rao JS. Matrix metalloproteinase-1 promotes prostate tumor growth and metastasis. Int J Oncol, 2008,32(4):757–765Google Scholar
  18. 18.
    Stamenkovic I. Matrix metalloproteinases in tumor invasion and metastasis. Semin Cancer Biol, 2000,10(6):415–433CrossRefGoogle Scholar
  19. 19.
    Kessenbrock K, Plaks V, Werb Z. Matrix metalloproteinases: regulators of the tumor microenvironment. Cell, 2010,141(1):52–67CrossRefGoogle Scholar
  20. 20.
    Weiss KR, Cooper GM, Jadlowiec JA, et al. VEGF and BMP expression in mouse osteosarcoma cells. Clin Orthop Relat Res, 2006,450:111–117CrossRefGoogle Scholar
  21. 21.
    Hornebeck W, Lambert E, Petitfrère E, et al. Beneficial and detrimental influences of tissue inhibitor of metalloproteinase-1 (TIMP-1) in tumor progression. Biochimie, 2005,87(3–4):377–383CrossRefGoogle Scholar
  22. 22.
    Salvatore V, Focaroli S, Teti G, et al. Changes in the gene expression of co-cultured human fibroblast cells and osteosarcoma cells: the role of microenvironment. Oncotarget, 2015,6(30):28988–28998CrossRefGoogle Scholar
  23. 23.
    Olguin HC, Olwin BB. Pax-7 up-regulation inhibits myogenesis and cell cycle progression in satellite cells: a potential mechanism for self-renewal. Dev Biol, 2004, 275(2):375–388CrossRefGoogle Scholar
  24. 24.
    Wang W, Pan H, Murray K, et al. Matrix metalloproteinase-1 promotes muscle cell migration and differentiation. Am J Pathol, 2009,174(2):541–549CrossRefGoogle Scholar
  25. 25.
    Kessenbrock K, Wang CY, Werb Z. Matrix metalloproteinases in stem cell regulation and cancer. Matrix Biol, 2015,44–46:184–190CrossRefGoogle Scholar
  26. 26.
    Cargnello M, Tcherkezian J, Roux PP. The expanding role of mTOR in cancer cell growth and proliferation. Mutagenesis, 2015,30(2):169–176CrossRefGoogle Scholar
  27. 27.
    Shay G, Lynch CC, Fingleton B. Moving targets: Emerging roles for MMPs in cancer progression and metastasis. Matrix Biol, 2015,44–46:200–206CrossRefGoogle Scholar
  28. 28.
    Willis AL, Sabeh F, Li XY, et al. Extracellular matrix determinants and the regulation of cancer cell invasion stratagems. J Microsc, 2013,251(3):250–260CrossRefGoogle Scholar
  29. 29.
    Fan HX, Chen Y, Ni BX, et al. Expression of MMP-1/PAR-1 and patterns of invasion in oral squamous cell carcinoma as potential prognostic markers. Onco Targets Ther, 2015,8:1619–1626Google Scholar
  30. 30.
    Kimura R, Ishikawa C, Rokkaku T, et al. Phosphorylated c-Jun and Fra-1 induce matrix metalloproteinase-1 and thereby regulate invasion activity of 143B osteosarcoma cells. Biochim Biophys Acta, 2011,1813(8):1543–1553CrossRefGoogle Scholar
  31. 31.
    Lou N, Wang Y, Sun D, et al. Isolation of stem-like cells from human MG-63 osteosarcoma cells using limiting dilution in combination with vincristine selection. Indian J Biochem Biophys, 2010,47(6):340–347Google Scholar
  32. 32.
    Tian J, Li X, Si M, et al. CD271+ osteosarcoma cells display stem-like properties. PLoS One, 2014,9(6):e98549CrossRefGoogle Scholar
  33. 33.
    Husmann K, Arlt MJ, Muff R, et al. Matrix metalloproteinase 1 promotes tumor formation and lung metastasis in an intratibial injection osteosarcoma mouse model. Biochim Biophys Acta, 2013,1832(2):347–354CrossRefGoogle Scholar

Copyright information

© Huazhong University of Science and Technology 2018

Authors and Affiliations

  • Man-li Tang
    • 1
  • Xiang-jun Bai
    • 1
  • Yong Li
    • 2
    • 3
    • 4
  • Xiao-jing Dai
    • 5
  • Fan Yang
    • 1
    • 2
    • 3
    • 4
    Email author
  1. 1.Department of Traumatic Surgery, Tongji Hospital, Tongji Medical CollegeHuazhong University of Science and TechnologyWuhanChina
  2. 2.Department of Orthopaedic Surgery, McGovern Medical Schoolthe University of Texas Health Science Center at Houston (UTHealth)HoustonUSA
  3. 3.Center for Stem Cell and Regenerative MedicineUTHealthHoustonUSA
  4. 4.Center for Regenerative Sports MedicineSteadman Philippon Research InstituteVailUSA
  5. 5.Department of Pharmacology and Toxicology, College of Pharmacythe University of HoustonHoustonUSA

Personalised recommendations