Exp Mol Med.  2010 Jan;42(1):38-46. 10.3858/emm.2010.42.1.004.

Induction of tissue inhibitor of matrix metalloproteinase-2 by cholesterol depletion leads to the conversion of proMMP-2 into active MMP-2 in human dermal fibroblasts

• Kim S
• Oh JH
• Lee Y
• Lee J
• Cho KH
• Chung JH

• KMID: 946668
• DOI: http://doi.org/10.3858/emm.2010.42.1.004

Abstract

Cholesterol is one of major components of cell membrane and plays a role in vesicular trafficking and cellular signaling. We investigated the effects of cholesterol on matrix metalloproteinase-2 (MMP-2) activation in human dermal fibroblasts. We found that tissue inhibitor of matrix metalloproteinase-2 (TIMP-2) expression and active form MMP-2 (64 kD) were dose-dependently increased by methyl-beta-cyclodextrin (MbetaCD), a cholesterol depletion agent. In contrast, cholesterol depletion-induced TIMP-2 expression and MMP-2 activation were suppressed by cholesterol repletion. Then we investigated the regulatory mechanism of TIMP-2 expression by cholesterol depletion. We found that the phosphorylation of JNK as well as ERK was significantly increased by cholesterol depletion. Moreover, cholesterol depletion-induced TIMP-2 expression and MMP-2 activation was significantly decreased by MEK inhibitor U0126, and JNK inhibitor SP600125, respectively. While a low dose of recombinant TIMP-2 (100 ng/ml) increased the level of active MMP-2 (64 kD), the high dose of TIMP-2 (> or = 200 ng/ml) decreased the level of active MMP-2 (64 kD). Taken together, we suggest that the induction of TIMP-2 by cholesterol depletion leads to the conversion of proMMP-2 (72 kD) into active MMP-2 (64 kD) in human dermal fibroblasts.

Keyword

cholesterol; extracellular signal-regulated MAP kinases; JNK mitogen-activated protein kinases; matrix metalloproteinase 2; tissue inhibitor of metalloproteinase-2

MeSH Terms

Anthracenes/pharmacology
Butadienes/pharmacology
Cells, Cultured
Child
Child, Preschool
Cholesterol/metabolism/*physiology
Cyclodextrins/pharmacology
Enzyme Inhibitors/pharmacology
Extracellular Signal-Regulated MAP Kinases/antagonists & inhibitors/physiology
Fibroblasts/*drug effects/*metabolism/ultrastructure
Humans
Immunoblotting
Immunoprecipitation
JNK Mitogen-Activated Protein Kinases/antagonists & inhibitors/physiology
Matrix Metalloproteinase 2/*metabolism
Microscopy, Electron, Transmission
Nitriles/pharmacology
Tissue Inhibitor of Metalloproteinase-2/*metabolism