HER2/neu动物模型的建立和免疫学鉴定

Establishment and evaluation of animal tumor model expressing HER2/neu

目的 建立表达人HER2 neu原癌基因的动物肿瘤模型并用免疫学方法加以鉴定。方法 将重组HER2 neu原癌基因真核表达载体转染P815细胞 ,有限稀释法筛选出稳定表达HER2 neu基因的P815肿瘤细胞株 ;用Westernblot方法鉴定HER2 neu基因在P815细胞中的表达。在DBA 2小鼠体内建立表达HER2抗原的肿瘤动物模型 ,再通过表达HER2 neu基因的重组质粒免疫小鼠 ,诱导小鼠产生特异性HER2 neu基因特异的CTL应答 ,从免疫学方面鉴定HER2 neu动物肿瘤模型建立成功。结果 在体外获得了稳定表达HER2 neu基因的P815细胞株 ;免疫小鼠脾细胞中诱导出特异性CTL。结论 本研究成功建立了HER2 neu动物肿瘤模型 ,HER2

Objective To establish animal tumor model expressing human oncogene HER2/neu and to evaluate the model by immunological method. Methods The recombinant eukaryotic plasmid expressing HER2/neu gene was transfected into P815 tumor cells. The tumor cell line P815 expressing HER2/neu gene stably was screened by limiting dilution method. The expression of target gene in host cells was determined by Western blotting. Following the establishment of animal tumor model expressing HER2 antigen in DBA/2 mice, tumor specific response was induced in the spleen cells of DBA/2 mice which were inoculated with recombinant plasmids. Results P815 cell line expressing HER2/neu oncogene stably was obtained. HER2/neu specific CTLs could be induced from immunized DBA/2 mice. Conclusion HER2/neu expressing tumor animal model is established successfully and the in vivo immune response can be induced after immunization with HER2/neu genetic vaccine.