表达谱基因芯片筛选烧伤后增生性瘢痕相关基因的研究

Study on differently expressed genes in postburn hypertrophic scar by cDNA microarray

目的 应用基因芯片技术从基因水平初步了解烧伤后增生性瘢痕形成的机制。 方法 按一步法抽提 3例烧伤患者的增生性瘢痕及其自身正常皮肤组织的总RNA ,纯化mRNA ;将40 96种人类基因PCR产物用CartesianPixsys 75 0 0点样仪微矩阵列点样于化学涂层的载玻片上 ,制成基因芯片 ;将等量的增生性瘢痕和患者自身正常皮肤组织mRNA分别逆转录合成荧光标记的cDNA混合物探针 ,与上述基因芯片杂交。经严格洗片后 ,用SanArray 30 0 0扫描仪扫描芯片荧光信号图像 ,计算机分析后比较两种组织中差异表达的基因。 结果 在 40 96种基因中 ,患者的增生性瘢痕及其自身正常皮肤组织间存在差异表达基因。在所检测的 3例临床标本中 ,共有差异表达基因 12 8条。 结论 包括细胞凋亡基因、免疫相关基因、细胞骨架和运动基因、原癌基因和抑癌基因。

Objective To screen the differently expressed genes between postburn hypertrophic scar and normal skin tissue using cDNA microarray. Methods The total RNAs from 3 cases were isolated from the tissues and purified to mRNAs by oligotex. The PCR products of 4 096 genes were spotted onto a chemical substance coated glass plate in array using Cartesian Pixsys 7 500. DNA was fixed onto the glass plate after series of treatments. Both mRNAs from the postburn hypertrophic scar and normal skin tissue were reversely transcribed to cDNAs with the incorporation of fluorescent dUTP to prepare the hybridization probes. The mixed probes were hybridized to the cDNA microarray. After high stringent washing, the cDNA microarray scanned for the fluorescent signals and showed differences between 2 tissues. Results Among 4 096 target genes, the expression level of 128 genes differed in 3 cases between the postburn hypertrophic scar and normal skin tissue. Conclusions Genes including the apoptotic genes, immune related genes, cellular signal and transferring genes take part in the development of postburn hypertrophic scar.