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. 2002 Jun 28;16(4):178–186. doi: 10.1002/jcla.10038

Decorin transfection in human mesangial cells downregulates genes playing a role in the progression of fibrosis

Antonia Costacurta 1,2,✉, Giovanna Priante 1, Angela D'Angelo 1, Luigi Chieco‐Bianchi 2, Salvatore Cantaro 3
PMCID: PMC6807966  PMID: 12112390

Abstract

The proteoglycan decorin inhibits TGF‐β; therefore, it could antagonize progression of fibrotic diseases associated with activation of TGF‐β1. The effect of decorin transfection in human mesangial cells (HMCs) on the expression of genes related to kidney fibrosis was investigated. HMCs, isolated from glomeruli of healthy portions of human kidneys removed due to carcinoma, were histochemically typed. Decorin cDNA cloned in a eukaryotic expression vector was transfected into HMCs. Gene expression of fibrogenetic cytokines and fibrotic proteins TGF‐β1, PDGF‐β, α1 collagen type IV, α1 collagen type I, fibronectin, and tenascin was analyzed, by reverse transcription polymerase chain reaction (RT‐PCR), 24 hr after transfection. Immunoblotting analysis of protein extracts using anti‐decorin IgG, revealed a positive signal of about 52 MDa, corresponding to the molecular weight of decorin, in cultures transfected with the decorin gene. Decorin mRNA increased about 12 times in cultures transfected with the construct pCR3.1‐Deco. Cells with increased decorin synthesis showed a 61% decrease of TGF‐β1 mRNA, a 71% reduction of α1 collagen type IV mRNA, and a 29% reduction of fibronectin mRNA. This study is the first to investigate decorin transfection into human mesangial cells, and supports the use of the decorin gene to control the progression of glomerular and interstitial fibrosis in kidney diseases. © 2002 Wiley‐Liss, Inc.

Keywords: human mesangial cell transfection, RT‐PCR, chronic renal fibrosis, gene therapy, α1 collagen type IV, fibronectin, TGF‐β1

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