敲减PTGS2对皮肤成纤维细胞基因表达谱的作用

Gene expression profiling of skin fibroblasts with silenced PTGS2

目的研究敲减PTGS2对成纤维细胞全基因组表达谱的影响,在基因水平上探索防治瘢痕疙瘩的新途径。方法运用RNAi干扰正常皮肤成纤维细胞前列腺素内过氧化物合酶2(PTGS2)基因的表达,利用real time RT-PCR验证siRNA沉默效果;应用全基因组芯片检测基因表达谱变化。结果成纤维细胞的PTGS2基因经siRNA干扰后,其mRNA表达水平明显下调;全基因组芯片表达谱检测到的差异表达基因,按1.5倍差异共189个(115个上调,74个下调),按2倍差异共14个(9个上调,5个下调);基因表达谱的变化与瘢痕疙瘩基因表达谱的变化相吻合,可能促使正常皮肤成纤维细胞向瘢痕疙瘩方向进展。结论检测到与PTGS2基因相关的、在瘢痕疙瘩形成中可能共同发挥作用的相关基因,证明PTGS2与瘢痕疙瘩的发病机制有着密切的关系,为治疗瘢痕疙瘩提供了一个潜在的候选靶点。

Objective This study is to elucidate the pathogenesis of keloid from the aspect of PTGS2 alteration and to find out potential targets for therapy.Methods Skin fibroblasts,the major inductive cell for keloid formation,were transfected with a recombinant PTGS2 siRNA expression vector.Real-time RT-PCR was used to confirm the knock down efficacy of PTGS2 siRNA.We performed gene expression analysis of the fibroblasts transfected with PTGS2 siRNA expression vector in comparison with the fibroblasts transfected with control vector using cDNA microarray chip which comprised of 21,552 clones.Different expression genes were annotated with literature mining method.Results The results of real-time RT-PCR indicated that recombinant PTGS2 siRNA expression vector could significantly knock down the expression of PTGS2 gene.We observed differential regulation of approximately 189 genes of the 10,682 efficient genes data in cDNA microarray,according to the data-filter-standard of 1.5 fold, of which 115 genes were up-regulated and 74 genes were down-regulated.Fourteen genes were differently expressed in transfected fibroblast and in control,9 genes were up-regulated and 5 genes were down-regulated.ConclusionsThe results suggest that PTGS2 is one of the key alterations responsible for keloid formation and that PTGS2 provides a potential target for therapy of keloid.