不同截短型癌基因AEG-1启动子活性分析  

The activity of differently truncated oncogene AEG-1 promoters

作　　者：代瑛[1] 龙敏[1] 张喆[1] 刘丽[1] 张惠中[1] 董轲[1] Dai Ying Long Min Zhang Zhe Liu Li Zhang Huizhong Dong Ke(Department of Clinical Research and Diagnosis, Tangdu Hospital, Fourth Military Medical University, Shaanxi Xi＇an 710038, China.)

机构地区：[1]第四军医大学唐都医院临床实验与检验科,陕西西安710038

出　　处：《现代肿瘤医学》2016年第23期3691-3696,共6页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:81201525)

摘　　要：目的:分析不同截短型癌基因AEG-1启动子活性,证实与E6/E7表达相关的转录调控因子对AEG-1启动子活性的影响。方法:以宫颈癌He La细胞基因组DNA为模版,扩增AEG-1启动子全长序列(-2710/-491),并应用本实验室改造的启动子活性研究专用绿色荧光蛋白报告载体构建p GL3-basicEGFP/AEG-1质粒,通过转染He La细胞及血管内皮细胞ECV304检测启动子活性。同时,根据AEG-1启动子序列上与E6/E7作用相关的转录调控因子结合位点位置,设计不同截短型AEG-1启动子序列引物,应用p GL3-basic-EGFP载体构建不同截短型AEG-1启动子载体,并分别转染细胞检测启动子活性。结果:AEG-1启动子全长序列p GL3-basic-EGFP/AEG-1载体转染He La细胞可见明显绿色荧光蛋白表达,而在血管内皮细胞ECV304中表达为痕量。包含不同转录调控因子C-Myc、SP1、NF-κB、E_2F结合位点的不同截短型AEG-1启动子载体在肿瘤细胞中有活性但存在差异。结论:AEG-1启动子为肿瘤特异性启动子,含有与E6/E7表达相关的转录调控因子NF-κB、E_2F结合位点的启动子序列为影响AEG-1基因启动子活性的关键区域。Objective： To analyze the activity of truncated oncogene AEG- 1 promoter and to confirm the effects of E6 / E7 related transcription regulatory factors on the AEG- 1 promoter activity. Methods： The full length sequences of AEG- 1 gene promoter（- 2710 /- 491） were amplified from He La cell genome DNA,and then cloned into p GL3- basic- EGFP vectors to constructe the promoter report plasmid of p GL3- basic- EGFP / AEG- 1 that was then transfected into He La and ECV304 cells to detect the AEG- 1 promoter tumor specific activity. Meanwhile,differently truncated region of AEG- 1 promoters were designed according to the transcriptional regulatory factors position on promoter sequences,and the activity were separately detected with the method described as above. Results： Strong green fluorescence could be observed in He La cells,but not in ECV304 cells when transfected with p GL3- basic-EGFP / AEG- 1 vector. The activities for differently truncated region of AEG- 1 promoters sequences with transcriptional regulation factors C- Myc,SP1,NF- κB,E2F binding sites had significant difference signals of green fluorescence. Conclusion： AEG- 1 gene promoter is a tumor specific activity promoter. The promoter sequences which have transcriptional regulation factor NF- κB,E2F,all of which are related to E6 / E7 function influences the activity of the promoter sequences.

关 键 词：AEG-1启动子 C-MYC SP1 NF-κB E2F 

分 类 号：R730.231[医药卫生—肿瘤]