论文部分内容阅读
目的:对构建的胃癌相关下调基因GDDR(Gastric Dramatic Down-related)转基因小鼠进行鉴定及功能初步检测,为深一步研究GDDR的生物学功能提供可靠的动物模型。方法:对GDDR转基因和野生对照(未经过基因修饰)小鼠胃组织使用实时定量PCR检测其基因转录情况,western blot和免疫组化检测其组织蛋白表达水平及表达部位。通过胃组织进行糖原染色和胃粘膜表面主要糖蛋白MUC5AC行免疫组化染色对小鼠表型进行初步分析。结果:实时定量PCR、western blot和免疫组化结果均显示在GDDR转基因小鼠中胃粘膜GDDR的表达水平明显高于野生组。糖原染色结果显示GDDR转基因小鼠的粘液分泌及粘液厚度高于正常对照组,糖蛋白MUC5AC的表达也显著强于野生组。结论:经过鉴定GDDR转基因小鼠的外源性基因GDDR得到成功表达。通过对GDDR转基因小鼠表型的初步分析,提示GDDR可能参与胃粘膜粘液层的组成和增加粘液分泌从而发挥着保护胃粘膜的作用。
OBJECTIVE: To identify and characterize the gastric cancer related gene GDDR (Gastric Dramatic Down-related) transgenic mice and provide a reliable animal model for further study on the biological function of GDDR. Methods: The gene transcription of GDDR transgenic mice and wild control mice (without genetic modification) were detected by real-time quantitative PCR. Western blot and immunohistochemistry were used to detect the expression of the protein and its expression site. The phenotypes of mice were analyzed by glycogen staining and MUC5AC, the major glycoprotein on gastric mucosa. Results: The results of real-time quantitative PCR, western blot and immunohistochemistry showed that the expression of GDDR in GDDR transgenic mice was significantly higher than that in wild-type mice. Glycogen staining showed that the mucus secretion and mucus thickness of GDDR transgenic mice were higher than that of the normal control group, and the expression of glycoprotein MUC5AC was also significantly stronger than that of the wild group. Conclusion: GDDR gene was successfully expressed in GDDR transgenic mice. Preliminary analysis of the GDDR transgenic mouse phenotype suggests that GDDR may play a role in protecting the gastric mucosa by participating in the composition of gastric mucosal layers and increasing mucus secretion.