增生性瘢痕与正常皮肤差异基因的筛选

Differentially expressed genes of hypertrophic scar and normal skin

目的 用人类基因制作的芯片 ,分析增生性瘢痕与正常皮肤组织之间基因表达的差异 ,筛选瘢痕形成过程中表达显著的差异基因。方法 选择 4例烧伤后 6个月、瘢痕严重增生挛缩的患者 ,取增生性瘢痕与自体正常皮肤进行对照 ,分成 4组 ,提取各个标本的总DNA与RNA ,制成荧光标记的cDNA探针 ,与含 4 0 0 0个人类靶基因的芯片 (15 0 0个为已知基因 ,2 5 0 0个为未报道或未知功能的基因 )杂交 ,经扫描 ,生物信息学分析 ,比较两种组织间的差异表达。结果 增生性瘢痕与正常皮肤的表达差异的基因在 4个组间仅出现 1次的有 378～ 4 5 1个 ,重复 2次出现的有 2 0 3～ 30 1个 ,3例标本中均有显著差异表达基因数为 114～ 15 2个 ,4例标本中均出现差异的基因则有 97个 ,其中以上调为主 ,涉及 13大类基因 ,包括抑癌与原癌基因 ,细胞凋亡 ,细胞骨架与运动蛋白 ,细胞周期类蛋白等 ,有些为已知基因 ,有些仍未知功能。结论 增生性瘢痕与皮肤的基因图谱存在差别 ,这些基因可能参与了增生性瘢痕的形成过程。

Objective To screen the differentially expressed genes between hypertrophic scar of burn and normal skin using gene microarray, to find out the genes of significantly different-expression and to analyse the roles of them in development of hypertrophic scar. Methods The total DNA and RNA from 4 hypertrophic scar samples and 4 normal skin were isolated and purified to mRNA by oligotex. They were reversely transcribed to cDNA with the incorporation of fluorescent dTUP to prepare the hybridization probes. Then, the mixed probes were hybridized to the cDNA chip and scanned for the signals and found differences between scar and normal skin. Results Among 4000 target genes, there were 378～451 genes once and 114～152 third times significantly different between hypertrophic scar and normal skin and 97 different-expressed genes in all 4 cases. Conclusions There are differences of gene expression between hypertrophic scar and normal skin screened by microarray. As the scanned cases increase, the common differentially-expressed genes become less and less, which may involve in the development of hypertrophic scar.