TY - JOUR
T1 - Up-regulation of tension-related proteins in keloids: Knockdown of hsp27, α2β1-integrin, and pai-2 shows convincing reduction of extracellular matrix production
AU - Suarez, Edna
AU - Syed, Farhatullah
AU - Alonso-Rasgado, Teresa
AU - Mandal, Parthasarathi
AU - Bayat, Ardeshir
AU - Suarez Pozos, Edna
PY - 2013/2
Y1 - 2013/2
N2 - BACKGROUND: Keloid disease is a fibroproliferative disorder, with an ill-defined treatment that is characterized by excessive extracellular matrix deposition. Mechanical tension promotes deposition of extracellular matrix and overexpression of tension-related proteins, which is associated with keloid disease. The aim of this study was to investigate the effect of tension-related proteins on extracellular matrix steady-state synthesis in primary keloid fibroblasts. METHODS: Keloid fibroblasts (n = 10) and normal skin (n = 4) fibroblast cultures were established from passages 0 to 3. A panel of 21 tension-related genes from microarray data were assessed at mRNA (quantitative reverse-transcriptase polymerase chain reaction) and protein (in-cell Western blotting) levels. Three genes were significantly altered in keloid tissue and fibroblasts, and their functional role was assessed using siRNA knockdown. RESULTS: Hsp27, α2β1-integrin, and PAI-2 were significantly up-regulated (p <0.05)in keloid tissue and fibroblasts compared with normal skin. Hsp27, α2β1-integrin, and PAI-2 expression was inhibited by RNA interference. Both the mRNA and protein levels of Hsp27, α2β1- integrin, and PAI-2 significantly decreased (p <0.05) in keloid fibroblasts at 48 hours after transfection. After down-regulation of Hsp27, α2β1-integrin, and PAI-2, the expression of intracellular extracellular matrix was significantly reduced (p <0.05). Water-soluble tetrazolium salt-1 assay showed that transfection of Hsp27, α2β1- integrin, and PAI-2 did not influence the viability/metabolic activity of keloid fibroblasts. CONCLUSIONS: This study demonstrates overexpression of key tension-related proteins in keloid tissue and keloid fibroblasts. Knockdown of Hsp27, PAI-2, and α2β1-integrin by RNA interference attenuates the expression of mRNA and protein levels and certain other extracellular matrix molecules. Copyright © 2013 by the American Society of Plastic Surgeons.
AB - BACKGROUND: Keloid disease is a fibroproliferative disorder, with an ill-defined treatment that is characterized by excessive extracellular matrix deposition. Mechanical tension promotes deposition of extracellular matrix and overexpression of tension-related proteins, which is associated with keloid disease. The aim of this study was to investigate the effect of tension-related proteins on extracellular matrix steady-state synthesis in primary keloid fibroblasts. METHODS: Keloid fibroblasts (n = 10) and normal skin (n = 4) fibroblast cultures were established from passages 0 to 3. A panel of 21 tension-related genes from microarray data were assessed at mRNA (quantitative reverse-transcriptase polymerase chain reaction) and protein (in-cell Western blotting) levels. Three genes were significantly altered in keloid tissue and fibroblasts, and their functional role was assessed using siRNA knockdown. RESULTS: Hsp27, α2β1-integrin, and PAI-2 were significantly up-regulated (p <0.05)in keloid tissue and fibroblasts compared with normal skin. Hsp27, α2β1-integrin, and PAI-2 expression was inhibited by RNA interference. Both the mRNA and protein levels of Hsp27, α2β1- integrin, and PAI-2 significantly decreased (p <0.05) in keloid fibroblasts at 48 hours after transfection. After down-regulation of Hsp27, α2β1-integrin, and PAI-2, the expression of intracellular extracellular matrix was significantly reduced (p <0.05). Water-soluble tetrazolium salt-1 assay showed that transfection of Hsp27, α2β1- integrin, and PAI-2 did not influence the viability/metabolic activity of keloid fibroblasts. CONCLUSIONS: This study demonstrates overexpression of key tension-related proteins in keloid tissue and keloid fibroblasts. Knockdown of Hsp27, PAI-2, and α2β1-integrin by RNA interference attenuates the expression of mRNA and protein levels and certain other extracellular matrix molecules. Copyright © 2013 by the American Society of Plastic Surgeons.
U2 - 10.1097/PRS.0b013e3182789b2b
DO - 10.1097/PRS.0b013e3182789b2b
M3 - Article
C2 - 23358011
SN - 0032-1052
VL - 131
JO - Plastic and Reconstructive Surgery
JF - Plastic and Reconstructive Surgery
IS - 2
ER -