Chen Zhenwei, Liu Yin, Xiao Minqin, Xiao Hong, Qiu Wei, Zhao Yanan
Department of Plastic Surgery, Second Affiliated Hospital, Kunming Medical University, Kunming Yunnan, 650101, P. R. China.
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2016 Jul 8;30(7):871-875. doi: 10.7507/1002-1892.20160177.
?To explore if Smad7 protein can inhibit growth of keloids by observing the gene and protein expressions of Smad7, collagen type I, and collagen type Ⅲ and cell proliferation after over-expression vectors of Smad7 transfecting keloid fibroblasts (KFb).
?Fibroblasts were acquired from 10 male patient with keloids at the age of 20 to 25 years. After in vitro culture, KFb were divided into 3 groups: untransfected group (group A), pcDNA3.1 (-) transfected group (group B), and pcDNA3.1 (-) -smad7 transfected group (group C). The mRNA and protein expression levels of Smad7, collagen type I, and collagen type Ⅲ were detected by real-time fluorescence quantitative PCR and Western blot at 48 hours after transfection. The cell proliferation ability was detected by MTT assay at 24 hours after transfection.
?The relative expression levels of mRNA and protein of Smad7 in group C were significantly higher than those in group A and group B (P<0.01). The relative expression levels of mRNA and protein of collagen type I and collagen type Ⅲ in group C were significantly lower than those in group A and group B (P<0.01). The relative expression levels of mRNA of collagen type I and collagen type Ⅲ in group B were significantly higher than those in group A (P<0.01); and the relative expression levels of proteins of Smad7, collagen type I, and collagen type Ⅲ were significantly lower than those in group A (P<0.01). The cell proliferation ability in group C was significantly lower than that in group A and group B at each time point by MTT assay (P<0.05), but no difference was found between group A and group B (P>0.05).
?Gene expressions of collagen type I, and collagen type Ⅲ and cell proliferation will be inhibited after KFb are transfected by over-expression vector of Smad7.
通过观察Smad7、Ⅰ型胶原蛋白、Ⅲ型胶原蛋白的基因和蛋白表达以及Smad7过表达载体转染瘢痕疙瘩成纤维细胞（KFb）后的细胞增殖情况，探讨Smad7蛋白是否能抑制瘢痕疙瘩的生长。
从10例年龄在20至25岁的男性瘢痕疙瘩患者中获取成纤维细胞。体外培养后，将KFb分为3组：未转染组（A组）、pcDNA3.1（-）转染组（B组）和pcDNA3.1（-）-smad7转染组（C组）。转染后48小时，采用实时荧光定量PCR和Western blot检测Smad7、Ⅰ型胶原蛋白和Ⅲ型胶原蛋白的mRNA和蛋白表达水平。转染后24小时，采用MTT法检测细胞增殖能力。
C组Smad7的mRNA和蛋白相对表达水平显著高于A组和B组（P<0.01）。C组Ⅰ型胶原蛋白和Ⅲ型胶原蛋白的mRNA和蛋白相对表达水平显著低于A组和B组（P<0.01）。B组Ⅰ型胶原蛋白和Ⅲ型胶原蛋白的mRNA相对表达水平显著高于A组（P<0.01）；而Smad7、Ⅰ型胶原蛋白和Ⅲ型胶原蛋白的蛋白相对表达水平显著低于A组（P<0.01）。MTT法检测结果显示，C组在各时间点的细胞增殖能力显著低于A组和B组（P<0.05），但A组和B组之间无差异（P>0.05）。
Smad7过表达载体转染KFb后，可抑制Ⅰ型胶原蛋白、Ⅲ型胶原蛋白的基因表达及细胞增殖。
