JCV病毒T抗原诱发晶状体上皮肿瘤动物模型建立  

The Establishment of Transgenic Mouse Model of Lens Epithelial Tumors Induced by JC Virus T Antigen

作　　者：王建平[1,2] 张梅英[3] 徐小燕[1,2] 王巍[1,2] 夏蒲[1,2] 郑志宏[3] 王禄增[3] 高野康雄 郑华川[1,2] 

机构地区：[1]中国医科大学生化与分子生物学教研室 [2]中国医科大学病理与病理生理研究所 [3]中国医科大学实验动物中心,沈阳110001 [4]神奈川县立癌中心临床研究所,日本横滨241-0815

出　　处：《中国医科大学学报》2011年第1期27-29,共3页Journal of China Medical University

基　　金：辽宁省百千万人才工程资助项目(2008921066);教育部归国人员科研启动基金(2009-03);人事部留学人员科技活动择优资助项目(2008-01);2009年沈阳市科学技术计划资助项目(1091175-1-00);沈阳人才专项基金(2009-01)

摘　　要：目的利用α-crystallin启动子构建晶状体上皮细胞中JCV T抗原表达转基因鼠,试图建立晶状体自发肿瘤动物模型。方法利用限制性内切酶BclI分离pBSK(pBluescript SK)-T抗原,BamH I同尾连接插入以pBSK为载体的α-crystallin启动子下游,通过NciI酶切电泳分离带有α-crystallin启动子的T抗原核酸序列后显微注射入C57BL/6J小鼠受精卵中。PCR检测转基因阳性鼠及JCV病毒T抗原拷贝数,大体和组织学观察眼球及病变组织,免疫组织化学观察T抗原、p53和β-catenin表达。结果成功构建α-crystallin JCV T抗原表达质粒,转基因阳性鼠中2号鼠(αAT-2)拷贝数最高,该鼠眼球于11个月表现出肿瘤样改变,肿瘤细胞中T抗原、p53和β-catenin呈强阳性表达。结论 JCV T抗原直接诱发晶状体上皮肿瘤动物模型建立为JCV T抗原嵌入所致上皮肿瘤提供了直接实验依据,为JCV所致上皮肿瘤发生和演进分子机理研究奠定坚实基础,同时为抗肿瘤药物筛选及基因治疗监控提供了良好的动物模型。Objective To establish the transgenic mouse model of spontaneous lens epithelial tumors using JCV T antigen initiated by α-crystallin promoter.Methods JCV T antigen was separated from pBSK（pBluescript SK）-T antigen plasmid by restriction enzyme BclI and compatibly ligated downstream α-crystallin promoter via BamH I.DNA fragments containing T antigen and α-crystallin promoter were separated from constructant by NciI and then microinjected into the fertilized eggs of C57BL/6J mice.The positive mice were screened by PCR and the comparative copies of T antigen were determined by real-time PCR.The lens of transgenic mice was grossly and histologically examined.The T antigen,p53 and β-catenin were examined in the lens lesions by immunohistochemistry.Results The α-crystallin-JCV T antigen expressing plasmid was successfully constructed.The T antigen copies of the No.2（αAT-2）transgenic mouse were highest in the 5 transgenic mice and displayed tumor-like lesion with the strong expression of T antigen,p53 and β-catenin proteins.Conclusions The transgenic tumor model provides a direct evidence for the oncogenic role of JCV T antigen insertation in the epithelial cells,a good animal model to investigate the oncogenic mechanisms of JCV T antigen,screen the anti-tumor reagents and monitor the gene therapy.

关 键 词：JC病毒 T抗原 转基因鼠 晶状体 

分 类 号：R739.7[医药卫生—肿瘤]