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

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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.