Detection of unique gene in Hypertrophic scar (Keloid) fibroblast by mRNA differential display strangy.

增生性瘢痕(瘢痕疙瘩)可由深度皮肤损伤(如广泛的热损伤)形成，其特征是真皮中细胞外基质的积聚。皮损皮肤成纤维细胞被认为在疾病的发展中起着至关重要的作用。事实上，已经观察到几种细胞外基质基因在皮损成纤维细胞中异常表达，但其功能的变化机制尚不清楚。为了解决成纤维细胞变化的分子机制，我们采用了差异显示的mRNA，这是一种新的基因消减策略。提取3例增生性瘢痕患者皮损和非皮损真皮成纤维细胞总RNA。将Poly-A RNA转化为cDNA，用20对引物进行聚合酶链式反应扩增，然后在测序凝胶上进行分析。在受累成纤维细胞中检测到37个突变克隆，与未受累成纤维细胞相比。然后对其中的4个克隆进行测序。为了分析这些基因的细节，将进行全长克隆和组织原位杂交。这种方法可能会让我们发现与疾病性质相关的基因(S)。

Hypertrophic scar (Keloid) can form as a result of deep dermal injury, such as extensive thermal injury, and are characterized by an accumulation of the extracellular matrix in dermis. Lesional skin fibroblasts have been belived to play a crucial role in the disease development. Indeed, abnormal expresions of several extracellular matrix gene in lesional fibroblasts have been observed, however, the alternation mechinery of its function is still unclear. To address molecular mechanisms of change in fibroblasts, we employed a mRNA of differential display which is a novel gene subtraction strategy. Total RNA of cultured dermal fibroblasts derived from involved and uninvolved lesions of Hypertrophic scar patients (n=3) were extracted. Poly-A RNAs were converted into cDNA,amplified by a polymerase chain reaction using 20 sets of primers, then analyzed on a sequencing gel.37 altered clones were detected in involved fibroblasts as compared to uninvolve fibroblasts. 4 clones of them were then sequenced.To analyze details of these genes, full-length cloning and tissue in situ hybridization will be pursued. This approach may lead us to identify gene (s) associated with the nature of the disease.