摘要
目的:利用细胞角蛋白19(K19)启动子构建胃黏膜上皮细胞中JC病毒(John Cunningham virus,JCV)T抗原表达转基因鼠,试图建立胃黏膜自发肿瘤动物模型。方法:利用限制性内切酶NdeⅠ,酶切K19-JCV T抗原表达质粒,电泳分离带有K19启动子的T抗原核酸序列后,显微注射入C57BL/6J30只小鼠受精卵中。PCR检测转基因阳性鼠,组织学观察主要脏器的形态学表现及免疫组化检测T抗原表达。结果:显微注射K19-JCV T抗原(KT)DNA片段后成功获得11种转基因阳性鼠,其中2个月龄KT7的各脏器组织学检测显示,脑、肝、肾、食管、前胃、胃、小肠、大肠、胰腺、肺、心脏和脾组织未见异常形态学改变,JCV T抗原特异表达于胃黏膜上皮细胞和支气管上皮细胞的细胞核中,16个月龄的KT7转基因鼠13.3%(2/15)出现肺肿瘤和腺癌的病理学改变,T抗原定位于癌细胞细胞核中。结论:JCV T抗原直接诱发肺肿瘤动物模型的建立为JCV T抗原嵌入所致上皮肿瘤提供了直接的实验依据,为JCV所致上皮肿瘤发生和演进分子机制奠定基础,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 gastric epithelial tumors using JCV T antigen initiated by cytokeratin 19 promoter （K19）. METHODS： DNA fragments containing T antigen and K19 promoter were separated from constructant by Nde I and then microinjected into the fertilized eggs of C57BL/6J mice. The positive mice were screened by PCR antigen by targeting JCV T antigen. The organs of transgenic mice were histologically exam- ined with JCV T antigen immunostained. RESULTS：The positive mice for K19-JCV T antigen was successfully established by microinjection. There was no abnormal appearance of brain, liver, kidney, esophagus, prestomach, stomach, small intes- tine, large intestine, pancreas, lung, heart and spleen in 2-month KT7 mouse. The positivity of JCV T antigen was observed in the nucleus of gastric and bronchial epithelial cells. The 13.3% （2/15） of 16-month KT7 mice showed pulmonary adeno- ma or adenocarcinoma with T antigen positive in the nuclei of carcinoma cells. CONCLUSIONS：The transgenic tumor mod- el provides a direct evidence for the oncogenic role of JCV T antigen insertion 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.
出处
《中华肿瘤防治杂志》
CAS
北大核心
2014年第5期321-324,共4页
Chinese Journal of Cancer Prevention and Treatment
基金
国家自然科学基金(81172371)
沈阳市科技攻关项目(F11-264-1-10
F12-277-1-01)
关键词
JC病毒
T抗原
转基因鼠
肺肿瘤
JC virus
T antigen
transgenic mouse
lung neoplasms
