mRNA差异显示法筛选和克隆胃癌相关基因

Screening and cloning of gastric carcinoma-related gene by mRNA differential display

目的 :筛选和克隆胃印戒细胞癌与正常胃黏膜细胞差异表达基因片段。方法 :采用mRNA差异显示法 (mRNAdifferentialdisplayPCR ,DD PCR) ,对比胃印戒细胞癌组织与正常胃黏膜组织mRNA的表达差异 ,克隆胃印戒细胞癌差异片段 ,经过RNA的斑点杂交、测序以及序列分析和同源性比较。结果 :胃印戒细胞癌与正常胃黏膜中存在明显的基因表达差异 ,发现与胃癌相关的差异表达条带 16条 ,其中 7条为缺失表达条带 ,9条为过度表达条带。对其中 4条差异表达条带进行克隆、测序 ,经序列分析及同源性比较和RNA的斑点杂交 ,表明确认其中 1条为GenBank/BLAST中尚未收录的片段。结论 :胃印戒细胞癌的mRNA差异显示证明 。

OBJECTIVE:To isolate and clone gastric carcinoma related gene. METHODS:The specially expressed cDNA fragments of gastric carcinoma were screened by differential display polymerase chain reaction (DD-PCR) after extracting the total RNA from tissues of human gastric carcinoma and normal gastric mucosa, then cloned these fragments to plasmid TA,and subsequently underwent sequence analysis,homogenous comparing and RNA dot hybridization.RESULTS:Obvious differences in gene expression between gastric carcinoma and normal gastric mucosa were found in 16 fragments express,of which 9 fragments were overexpressed and 7 fragments default expressed in gastric carcinoma.Among them 4 fragments were cloned and by sequence analysis,homogenous comparing and dot hybridization,one of them showed novel .CONCLUSION:The specially expressed EST sequence might be the gastric carcinoma related oncogene and maybe play a certain role in the development of gastric carcinoma.