用差异显示反转录PCR银染技术研究小鼠前胃癌相关基因

A Study of Associated Genes in NIH mice's Pregastric Carcinoma by DDRT-PCR and Silver Staining

目的:以N-亚硝基-肌氨酸乙酯(NSEE)作为诱变剂建立NIH小鼠前胃癌动物模型,研究小鼠前胃癌差异表达基因,探索细胞癌变的分子机理.方法:采用mRNA差异显示(DDRT-PCR)、反向Northern点杂交、克隆、测序和生物信息学分析等方法,对其癌变组织中差异表达的基因进行系统的研究分析.结果:得到5条在小鼠前胃正常对照组织、癌变组织之间差异表达的cDNA片段.筛选后,对其中3个差异条带进行DNA序列测定,有1个与已知基因Tpt1高度同源,2个与同1基因片段AK085193.1高度同源.提示这2个基因与小鼠前胃癌的形成有关.

To study the molecular mechanism of the pregastric carcinogenesis, we used N-Nitrososarcosineethylester （NSEE） as an induced regent to make the animal models of NIH mice＇s pregastric carcinoma, and investigated differential expression of the genes between the normal and the carcinoma tissues. Methods： The mRNA differential display reverse transcription PCR, the reverse Northern blot, clone, sequence the cDNA bands, bioinformatics and so on, were used to scientifically analyze differential expression of the genes between the normal and the carcinoma tissues. Results： 5 cDNA fragments were found during the experiments. After screened, three of which were cloned and their DNA sequences were analyzed. One of them was highly homologous with Tptl gene. And the others were highly homologous with the fragment AK085193.1. This suggested that they were related with the carcinogenesis of mice＇s pregastric carcinoma.