Title: Effects of Tissue Inhibitor of Metalloproteinase-1 on Cell Proliferation,matrix Metalloproteinases(MMPs) and Extracellular matrix(ECM) Degradation in Fibroblast
Abstract: ObjectiveTo investigate the possible effects of tissue inhibitor of metalloproteinase-1(TIMP-1) on cell proliferation,matrix metalloproteinases(MMPs) and extracellular matrix(ECM) degradation in fibroblast.MethodsNIM3T3 cells transfected with recombinant mammalian expression retroviral vectors(pLXIN) carried sense human TIMP-1 cDNA(PTs) and antisense human TIMP-1 cDNA(PTas).Cell proliferation was measured by cell count.Type Ⅰ,Ⅲ and Ⅳ collagens and fibronectin(FN) content were tested in Cell culture supernatants by ELISA and MMP-2,MMP-9 content by Gelatin zymogram.Collagen-alpha 1 (Ⅳ) mRNA,MMP-2 mRNA,MMP-9 mRNA and TIMP-1 mRNA were analyzed by northern blot.ResultsProliferation of cultured NIH3T3 cells was enhanced in PTs transfected and was reduced in PTas transfected.A markedly decrease in expression of endogenous TIMP-1 mRNA in PTas transfected was confirmed by northern blot analysis.However,expressions of mRNA for collagen-alpha 1(Ⅳ),MMP-2,MMP-9 were not markedly changed.The concentrations of type Ⅰ,Ⅲ,Ⅳ collagnes and FN were markedly decreased in PTas transfected cells culture supernatants and markedly increased in PTs transfected cells culture supernatants.The contents of MMP-2,MMP-9,were markedly increased in PTas transfected cells culture supernatants and markedly decreased in PTs transfected cells culture supernatants.Conclusion These results suggested that antisense TIMP-1 mRNA suppress cell proliferation and the expression of endogenous TIMP-1,resulting in increase of ECM degradative activity of MMPs,hence it contributed to the degradation of ECM.
Publication Year: 2004
Publication Date: 2004-01-01
Language: en
Type: article
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