Osteoblast-derived matrix metalloproteinases (MMPs) are considered to play a crucial
role in bone formation and initiation of bone resorption by degrading the bone matrix.
MMP-2 is constitutively secreted in a latent zymogen by osteoblasts, and requires
the process of activation mediated by membrane-type matrix metalloproteinase-1 (MT1-MMP)/tissue
inhibitor of metalloproteinase (TIMP-2) complex in the cell surface. Bone is one target
tissue for progestins. In the present study, we observed the effects of progesterone
on proMMP-2 activation and MT1-MMP expression, and also TIMP-2 levels in osteoblastic
MG-63 cells. Gelatin zymograms and ELISA showed that progesterone have no effects
on proMMP-2 activation. Using Western immunoblot analysis, we unexpectedly found that
treatment with increasing doses of progesterone in MG-63 cells caused a dose-dependent
increase in expression of MT1-MMP protein, and after 48 h treatment, progesterone
at 10-8 M increased MT1-MMP protein level. Confocal immunohistochemistry analysis also confirmed
that progesterone induced MT1-MMP expression in MG-63 cells. The results of Northern
blot analysis showed that progesterone at 10-8 M increased MT1-MMP protein levels after 48 h treatment. We also found that TIMP-2
levels were undetectable in MG-63 cells. In conclusion, progesterone increases MT1-MMP
protein and mRNA levels in MG-63 cells, but has no effects on proMMP-2 activation,
which is partly attributable to the undetectable levels of tissue inhibitor of metalloproteinase-2
(TIMP-2). Our studies suggest that TIMP-2 is involved in proMMP-2 activation, and
regulation of MT1-MMP by progesterone may contribute to its actions on bone formation.
Key words:
Progesterone - MT1-MMP - proMMP-2 - TIMP-2
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Xianghang Luo
Institute of Endocrinology & Metabolism
The Second Affiliated Hospital of Hunan Medical University
Changsha, Hunan, 410011
PR China
Phone: + 86 (731) 55 50 254
Fax: + 86 (731) 55 33 525
Email: xianghangluo@21cn.com