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


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2018-10

AUTHORS

Man-li Tang, Xiang-jun Bai, Yong Li, Xiao-jing Dai, Fan Yang

ABSTRACT

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. More... »

PAGES

809-817

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s11596-018-1947-5

DOI

http://dx.doi.org/10.1007/s11596-018-1947-5

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1110953299

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/30594980


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