Effects of SiRNA-mediated Smad3 gene silence on the expression of fibronectin and collagen Ⅰ in human keloid fibroblasts

LIU Bo; et al

doi:10.13406/j.cnki.cyxb.2010.04.022

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Effects of SiRNA-mediated Smad3 gene silence on the expression of fibronectin and collagen Ⅰ in human keloid fibroblasts

更新时间：2024-01-17

- Effects of SiRNA-mediated Smad3 gene silence on the expression of fibronectin and collagen Ⅰ in human keloid fibroblasts
- Journal of Chongqing Medical University Vol. 35, Issue 4, Pages: 522-526(2010)
- 作者机构：
  
  重庆医科大学附属第一医院烧伤整形外科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.04.022
  CLC： R622
- Published：2010，
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[1]刘波,果磊.SiRNA-Smad3沉默人瘢痕疙瘩成纤维细胞中Smad3对纤维连接蛋白及Ⅰ型胶原表达的影响[J].重庆医科大学学报,2010,35(04):522-526.

LIU Bo, et al. Effects of SiRNA-mediated Smad3 gene silence on the expression of fibronectin and collagen Ⅰ in human keloid fibroblasts. [J]. Journal of Chongqing Medical University 35(4):522-526(2010)
[1]刘波,果磊.SiRNA-Smad3沉默人瘢痕疙瘩成纤维细胞中Smad3对纤维连接蛋白及Ⅰ型胶原表达的影响[J].重庆医科大学学报,2010,35(04):522-526. DOI： 10.13406/j.cnki.cyxb.2010.04.022.

LIU Bo, et al. Effects of SiRNA-mediated Smad3 gene silence on the expression of fibronectin and collagen Ⅰ in human keloid fibroblasts. [J]. Journal of Chongqing Medical University 35(4):522-526(2010) DOI： 10.13406/j.cnki.cyxb.2010.04.022.

摘要

目的:探讨SiRNA-Smad3沉默人瘢痕疙瘩成纤维细胞（Keloid fibroblast

KFB）内Smad3基因后

对KFB细胞内纤维接蛋白及Ⅰ型胶原表达水平的影响。方法:分别用针对人Smad3 mRNA基因的特异性干扰片段和无关干扰片段转染体外原培养的人KFB

经RT-PCR、Western blot、免疫组化、免疫荧光方法检测纤维连接蛋白（fibronectin）及Ⅰ型胶原(CollagenⅠ

Col等KFB细胞外基质的合成和分泌情况。结果:75nmol/L Smad3-siRNA特异性干扰片段转染KFB48h后KFB细胞内ColⅠ及合成和分泌较非特异性对照组及空白对照组明显降低。结论:Smad3-siRNA可特异性抑制KFB细胞中Smad3基因表达

并效抑制体外培养的人原代KFB合成和分泌ColⅠ及纤维连接蛋白（Fibronectin

FN）。

Abstract

Objectives:To explorer the influence of siRNA-Smad3-mediated Smad3 gene silence on the expression of fibronectin and collagenⅠin human keloid fibroblast(sKFB).Methods:One specific Smad3-siRNA fragment and another irrelevant siRNA fragment was respectively transfected into human primary KFB cells.RT-PCR

Western blot

immunohistochemisty and immunofluorescence were employed to evaluate the changes in either the expression or the secretion of both fibronectin and collagenⅠ.Restults:The expression and secretion of both the fibronectin and collagenⅠin KFB were inhibited significantly by the treatment of Smad3-siRNA for 48 hours at a final concentration of 75nmol/L as compared with those of the untreated or the irrelevant siRNA fragment treated group.Conculsion:Smad3-siRNA could be employed to specifically inhibit the Smad3 gene expression in KFB cells and to efficiently inhibit both the synthesis and secretion of fibronectin and collagenⅠ.

关键词

RNA干扰Smad3瘢痕疙瘩成纤维细胞

Keywords

RNA interferenceSmad3keloidfibroblast

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