【摘要】 目的研究宫颈癌中游离型HPV16中细胞特异性增强子(cell-type specific enhancer,CTSE)片段的变异对启动子P97活性的影响,探讨CTSE片段变异与游离型HPV16致癌机制的关系。方法分别构建含HPV16突变型及野生型CTSE片段的重组报道基因载体(pCAT3-Promoter)表达系统,利用脂质体将重组的报道基因载体转染HeLa细胞;ELISA法测定细胞中CAT蛋白表达量的变化,了解变异CTSE片段对启动子活性的影响。结果与无增强子的CAT报道质粒pCAT3-Promoter相比,插入HPV16 CTSE序列的pCAT3-Promoter重组报道质粒CAT蛋白表达量增加1.5~2.5倍;插入突变型CTSE序列的报道载体与插入野生型CTSE序列的报道载体相比,报道基因CAT蛋白表达量增加了约1.6倍。结论陕西地区宫颈癌组织中HPV16 YY1位点突变的CTSE序列可激活启动子活性,YY1位点的突变与游离型HPV16 DNA致宫颈癌有密切关系。更多还原

【Abstract】 Objective To investigate the influence of mutant episomal HPV16 CTSE on promoter activity. Methods Recombinant CAT reporter plasmids containing wild HPV- 16 CTSE( pCAT3- Promoter- wCTSE) and mutant episomal HPV16 CTSE( pCAT3- Promoter- mCTSE) were constructed with pCAT3- Promoter system. Functional analysis of the mutant CTSE and the wild- type CTSE was performed with CAT assay. Results 1. The recombinant plasmids showed a 1. 5- 2. 5 fold increase in comparison to the CAT expressed of the pCAT3- Promoter. 2. The recombinant plasmids pCAT- 2 showed a 1. 6 fold increase of the CAT expressed in comparison to pCAT- 1 containing the prototype HPV- 16 CTSE. Conclusions Our results indicated the activity of promoter was up- regulated by YY1 site mutation in this CTSE. The mechanism of episomal HPV- associated carcinogenesis in Shaanxi is closely associated with YY1 site mutation in LCR.更多还原