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[Recombinant MMP-14 catalytic domain inducing the apoptosis of human osteoblastic SaOS-2 cells]
Li-Juan Guo1, Xiang-Hang Luo, Xin Su
1Institute of Endocrinology and Metabolism, Second Xiangya Hospital, Central South University, Changsha 410011, China.
Objective:
To investigate the direct effects of recombinant matrix metalloproteinase-14 (MMP-14) catalytic domain (RMC) on cultured human osteoblastic SaOS-2 cells.
Methods:
Western Blotting was used to analyze the expression of MMP-14 in cultured media. The effect of RMC on activation of proMMP-2 was measured by gelatin zymograms analysis, and the influence of RMC on cells adhesion was analyzed as well. The apoptosis was detected by acidine orange/ethidium bromide staining and ELISA.
Results:
RMC activated proMMP-2 secreted into media by SaOS-2 cells, and this process was blocked by EDTA treatment. RMC inhibited the adhesion of SaOS-2 cells to immobilized Type I collagen in a dose-dependent manner, which was also abolished by EDTA. RMC induced SaOS-2 cells apoptosis in a dose-dependent manner, and apoptosis-inducing activity of MMP-14 catalytic domain was blocked by EDTA.
Conclusion:
Since adhesion of cells to extracellular matrix serves as a survival mechanism in osteoblasts, the catalytic activity of recombinant MMP-14 catalytic domain on matrix proteins contributes to its apoptosis-inducing activity.
Insights
Recombinant matrix metalloproteinase-14 (MMP-14) catalytic domain (RMC) activates proMMP-2 and induces apoptosis in osteoblast-like cells by inhibiting cell adhesion. This suggests MMP-14
Area of Science:
- Biochemistry
- Cell Biology
- Extracellular Matrix Biology
Context:
- Osteoblasts are crucial for bone health, relying on interactions with the extracellular matrix for survival.
- Matrix metalloproteinases (MMPs) play roles in tissue remodeling, but their specific effects on osteoblast function require further elucidation.
- Understanding MMP-14's direct impact on osteoblasts is key to deciphering bone homeostasis and pathology.
Purpose:
- To directly investigate the effects of the recombinant MMP-14 catalytic domain (RMC) on human osteoblastic SaOS-2 cells.
- To analyze RMC's influence on proMMP-2 activation, cell adhesion to Type I collagen, and apoptosis.
Summary:
- RMC activated proMMP-2 secreted by SaOS-2 cells, an effect inhibited by EDTA.
- RMC dose-dependently inhibited SaOS-2 cell adhesion to Type I collagen, also reversible by EDTA.
- RMC induced SaOS-2 cell apoptosis in a dose-dependent manner, with EDTA blocking this effect.
Impact:
- The catalytic activity of MMP-14 on matrix proteins contributes to apoptosis induction in osteoblasts.
- Cell adhesion to the extracellular matrix is a survival mechanism for osteoblasts.
- These findings highlight a novel mechanism by which MMP-14 may influence bone cell viability and function.
