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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
Article
  • 17 Downloads

Summary

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 

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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

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