【作者】 阳彦；

【导师】 卢光琇；

【作者基本信息】 中南大学 ， 遗传学， 2010， 硕士

【摘要】 目的：目前,肺癌已经成为威胁人类健康的第二大癌症,本实验通过构建整合外源基因SPC-neu-polyA的转基因小鼠,对其表型进行鉴定,旨在通过该转基因小鼠模型研究neu基因在肺癌发展中的作用以及肺炎与肺癌的关系。方法：本实验分为两大部分,第一部分为显微注射得到整合SPC-neu-polyA的转基因小鼠,通过PCR、Southern blot、Western blot、FISH、免疫组化对其进行鉴定,第二部分为转基因小鼠的表型鉴定,病理切片HE染色病理鉴定,Real-Time PCR鉴定炎症因子的表达、免疫组化鉴定炎症因子蛋白的表达。补充实验：免疫组化检测10例肺癌病人组织活检标本的neu蛋白表达和炎症因子的表达。结果：通过PCR、Southern blot、FISH、检测SPC-neu-polyA完整的整合到转基因小鼠的16号染色体上。Western blot检测到了转基因小鼠中有neu蛋白的表达,IHC实验显示neu蛋白在转基因阳性小鼠的肺泡和支气管上皮表达。病理切片HE染色发现转基因阳性鼠较正常小鼠有明显的炎症病变,并在年长小鼠中出现上皮细胞增生严重等癌前病变现象。免疫组化检测到10例肺癌病人肺组织活检标本均呈neu阳性或弱阳性,并伴有炎症因子表达。结论：从以上实验结果,SPC-neu-polyA转基因小鼠已建系成功,表型鉴定为其为炎症到癌前病变的炎症疾病动物模型。该动物模型可用于研究neu在肺癌发展中的作用和研究肺炎与肺癌的关系。更多还原

【Abstract】 Objective:Lung cancer has currently become the No. 2 killer in cancers threatening human health. Our study was to establish a transgenic mice model integrated with an exogenous SPC-neu-polyA construct, identify their phenotype, so to investigate how the neu gene work in lung cancer development and the relationship between pneumonia and lung cancer.Methods:Our study included two parts. First, The SPC-neu-polyA sequence was microinjected into mice to generate transgenic mice, which were identified by PCR, Southern blot, Western blot, FISH and immunohistochemistry. Second, the phenotype of transgenic mice was identified by histological HE staining, Real Time-PCR and IHC. Supplement expreiment:10 cases of lung cancer biopsy specimens were detected by IHC to analyze the neu protein expression and inflammation factor expression.Results:The results of PCR, Southern blot and FISH suggested that SPC-neu-polyA was completely integrated into the genome of transgenic mice, and located in the 16th Chromosome. The results of Western blot suggested that the neu protein was expressed in transgenic mice; IHC experiments suggested that neu protein was expressed in the transgenic mice’s alveolar and bronchial epithelium. Significant inflammatory changes were found in the transgenic mice but not in the normal mice by Pathology HE staining. Some transgenic mice showed epithelial cell hyperplasia, one of precancerous lesions. Immunohistochemistry detection of 10 cases of lung biopsy specimens of lung cancer patients showed neu-positive and the expression of inflammatory factors.Conclusions:From these results, SPC-neu-polyA transgenic mice had been successfully established. Their phenotypes included inflammatory changes and precancerous lesions, indicating they were inflammatory disease models. The animal model can be used to study the role of neu in lung cancer development and the relationship between pneumonia and lung cancer.更多还原