2003•Journal of Shanghai MedicaRequires access
Influence on the Growth and Changes of Several Phenotypes of Cultured Rat MsC Transfected by DCN Gene
Guo Mu-yi
Open publisher page 2 citations
Abstract
Purpose To study the growth and the changes of some phenotypes of cultured rat mesangial cell (MsC) transfected by decorin(DCN) gene in order to provide experimental basis for gene delivery to di-seased glomeruli of rat mesangial proliferative glomerulonephritis. Methods We applied MTT assay and FCM analysis for examining MsC growth,Northern blot and Western blot analyses for measuring the levels of transforming growth factor-β1(TGF-β1) mRNA and its protein,and ColⅣ mRNA expressions respectively,semi-quatification RT-PCR method for detecting the level of type-2 tissue inhibitor of metalloproteinase(TIMP-2) mRNA expression on MsC. Results The inhibited growth,increased G 0/G 1 phase percentage and decreased S phase percentage of MsC clones transfected by DCN gene were observed by MTT method and FCM analysis respectively,compared to that of the untransfected MsC.The levels of TGF-β1 mRNA and ColⅣ mRNA expressions and TGF-β1 protein synthesis were decreased by Northern blot and Western blot analyses respectively,and TIMP-2 mRNA expression level of the transfectants were decreased by RT-PCR,compared to that of the untrnsfected MsC. Conclusions DCN gene transfected to MsC can inhibit MsC growth and ECM synthesis in varying degrees.