基因芯片法筛选萎缩性胃炎相关的差异表达基因

Screening of differentially expressed genes associated with atrophic gastritis by high density cDNA microarrays

目的:建立大鼠萎缩性胃炎模型,用高密度基因芯片筛选萎缩性胃炎相关的差异表达基因.方法:采用热盐水、盐水、热水灌胃的方法建立大鼠萎缩性胃炎模型,并设立正常对照组、正常喂养组作为对照.提取标本的总RNA,定量后并反转录成cDNA,萎缩性胃炎标本标记上Cy5,正常对照标本标记上Cy3,与含有8248个大鼠基因的cDNA芯片进行杂交,常规洗片,扫描,分析杂交结果.结果:热盐水组灌胃12wk的大鼠经病理诊断已存在萎缩性胃炎的特征,盐水、热水组灌胃的大鼠在24wk时经病理诊断也存在萎缩性胃炎的特征;热盐水组与正常对照组之间的差异基因共有436个,145个基因在萎缩性病变组织中呈高表达,291个基因在正常组织中呈高表达;热水组与正常对照组之间的差异表达基因共有398个,98个基因在萎缩性病变组织中呈高表达,300个基因在正常组织中呈高表达;热盐水组与盐水组之间差异表达基因共有36个,其中23个在热盐水组中呈高表达,13个基因在盐水对照组中呈高表达.结论:成功地建立了大鼠萎缩性胃炎的模型;筛选出了与热盐水、盐水、热水诱发的萎缩性胃炎发生有关的差异表达基因.

AIM:To establish the rat model and to screen differentially expressed genes closely associated with atrophic gastritis. METHODS:The rat atrophic gastritis model was induced by intragastrically giving hot high salt water, hot water and high salt water. Normal raising group and normal control group were used as control. Total RNAs of specimens were extracted, quantified, and reversely transcripted into cDNA. cDNAs from atrophic gastritis were labeled with Cy5, and cDNAs from the normal control group were labeled with Cy3, which were mixed with cDNAs from the control group with equal quantity, then hybridized with cDNA chips containing 8 248 genes. Chips were washed, scanned and analyzed. RESULTS:There were atrophic lesions in rat gastric sinus confirmed by pathological examination in the hot high salt water group at the 12th week, but in the high salt water group and the hot water group, there were atrophic lesions at the 24th week. A total of 436 differentially expressed genes were identified by cDNA chip between the hot high salt water group and the normal control group, 145 genes were highly expressed in the hot high salt water group and 291 genes were highly expressed in the normal control group; 398 differentially expressed genes were identified in the hot water group and the normal control group, 98 genes were highly expressed in the hot water group, and 300 genes were highly expressed in the normal control group; and 36 differentially expressed genes were confirmed be-tween the hot high salt water group and the high salt water group, 23 genes were highly expressed in the hot high salt water group and 13 genes were highly expressed in the high salt water group. CONCLUSION:Rat atrophic gastritis models were established successfully, differentially expressed genes associated with atrophic gastritis induced respectively by hot high salt water, high salt water and hot water were identified by way of high density cDNA chip.