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Effect of heat shock protein 47 on collagen accumulation in keloid fibroblast cells
Author(s) -
Chen JJ.,
Zhao S.,
Cen Y.,
Liu X.X.,
Yu R.,
Wu D.M.
Publication year - 2007
Publication title -
british journal of dermatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.304
H-Index - 179
eISSN - 1365-2133
pISSN - 0007-0963
DOI - 10.1111/j.1365-2133.2007.07898.x
Subject(s) - keloid , fibroblast , transfection , microbiology and biotechnology , small hairpin rna , western blot , extracellular , extracellular matrix , wound healing , messenger rna , intracellular , heat shock protein , chemistry , biology , cell culture , rna , biochemistry , medicine , immunology , pathology , gene , genetics
Summary Background Keloid is characterized by excessive collagen accumulation, but the mechanism of keloid formation remains unknown, and none of the treatment modalities are consistently effective. Heat shock protein (HSP) 47, known as a collagen‐specific molecular chaperone, plays a critical role in collagen biosynthesis. Our previous research has demonstrated that HSP47 is highly expressed in keloid compared with normal skin tissues, which indicates that there might be a close relationship between overexpression of HSP47 and excessive collagen accumulation in keloid formation. Objectives To further investigate whether overexpression of HSP47 might promote excessive collagen deposition in keloid formation, we examined the alteration of intracellular and extracellular collagen expression, following inhibition of HSP47 expression in keloid fibroblast cells by the RNA interference technique. Methods Three constructed psiRNA‐hH1neo plasmids, carrying three pairs of related HSP47‐shRNA (small hairpin RNA), respectively, were transfected into keloid fibroblast cells and compared with three control groups. After transfection, the mRNA and protein expression of HSP47 and collagen type I were detected by quantitative real‐time polymerase chain reaction and Western blot; the content of extracellular secreting collagen was assessed by hydroxyproline assay; and the MTT [3‐(4, 5‐dimethylthiazolyl‐2)‐2, 5‐diphenyltetrazolium bromide] method was adopted to examine the proliferation of keloid fibroblast cells. Results Both the mRNA and protein levels of HSP47 in keloid fibroblast cells decreased dramatically 48 h after post‐transfection of three related HSP47‐shRNA plasmids, compared with control groups. Following the downregulation of HSP47, we found that the expression of intracellular and extracellular collagen was correspondingly reduced. On the other hand, the MTT assay showed that transfection of HSP47‐shRNA plasmids did not influence the growth of keloid fibroblast cells. Conclusions Combined with our previous histological results, we propose that overexpression of HSP47 in keloid fibroblast cells could induce excessive collagen accumulation by enhancing synthesis and secretion of collagen, which not only presents a possible mechanism of keloid formation, but also offers a therapeutic potential of RNA interference to HSP47 for the treatment of keloid and other fibroproliferative disorders.