应用抑制性消减杂交筛选大鼠不同程度肝纤维化的上调基因

Screening and Cloning Up-Regulated Expressed Genes in Differential Liver Fibrosis by Suppression Subtractive Hybridization Technique

目的应用抑制消减杂交技术,筛选大鼠正常肝脏组织、轻度肝纤维化和重度肝纤维化组织中差异表达的上调基因。方法利用抑制消减杂交技术,构建大鼠轻度肝纤维化和重度肝纤维化2个差异cDNA文库,并挑选36条上调显著的基因进行测序鉴定。结果获得1152个克隆,其中有1036个含有插入片段。挑选32个克隆测序,与GenBank数据库进行初步比较,其中28条与已知基因的部分序列高度同源。结论应用抑制消减杂交技术成功构建了轻度肝纤维化和重度肝纤维化差异表达基因的cDNA削减文库,为进一步研究肝纤维化发生、发展和逆转的机制奠定了理论基础。

Objective To clone and identify up-regulated expressed genes among normal hepatic tissues, gentle liver fibrosis tissues and serious liver fibrosis tissues by suppression subtractive hybridization technique. Methods Two cDNA subtractive libraries of genes differentially expressed among normal hepatic tissues, gentle liver fibrosis tissues and serious liver fibrosis tissues were constructed by suppression subtractive hybridization technique. Results One thousand one hundred and fifty-two clones were obtained. The amplified library contained 1 036 positive clones. Sequence analysis was＇ performed in 32 clones, and the full length sequences were obtained with bioinformatics method. Altogether 28 coding sequences were gotten. Conclusion Two cDNA subtractive libraries of genes differentially expressed among normal hepatic tissues, gentle liver fibrosis tissues and serious liver fibrosis tissues were constructed successfully. The present study laid a theoretic foundation for exploring the mechanism of liver fibrosis.