Analysis of differentially expressed genes in keloids and normal control skin / 解放军医学杂志

ABSTRACT

Objective To study the differentially expressed genes between keloids and normal control skin using cDNA microarray. Methods The PCR products of 8400 human genes were spotted on a chemical-material-coated-glass plate in array. Total RNAs were isolated from the freshly excised human keloids and normal control skin,and then were purified to mRNAs,which were reversely transcribed to cDNAs with the incorporations of fluorescent dUTP,for preparing the hybridization probes. The mixed probes were hybridized to the cDNA microarray. According to the results of cDNA microarray,three genes (NGF,TGF-?_ 1 and c-myc) were chosen to study their differential expression in keloids and normal control skin by RT-PCR. Results Among the 8400 target genes,there were 402 genes with different expression (4.75%),and they were mainly associated with extracellular matrix genes,cellular signal molecule genes and cellular skeleton molecule genes. Analysis of collagen related molecule and growth factor gene expression confirmed that our molecular data obtained by cDNA microarray were consistent with published biochemical and clinical observations of keloids. RT-PCR showed that the levels of gene expression of NGF,TGF-?_ 1 and c-myc were all higher in keloids than those in normal control skin. Conclusion Many genes were found to be involved in the formation of keloids. Further analysis of the obtained genes might contribute to reveal the molecular mechanism of keloids formation,in which NGF,TGF-?_ 1 and c-myc might play important roles.