宫颈癌中HPV16介导转录因子AP-2α调控ASPP1和ASPP2基因转录分子机制研究

High risk HPV16 is causally close to occurrence and development of cervcial carcinoma and interaction between viral oncoproteins and host cells is emphasized to be investigated on molecular carcinogenesis of HPV16.A scientific hypothesis is presented based on our results suggested by previous laboratory experiments and it might be a new molecular mechanism that HPV16 could mediate transcription factor AP-2α to regulate transcription of ASPP1 and ASPP2 genes, aopotosis stimulating P53 protein family members. Firstly, the relationship between HPV16 and ASPP1/2, AP-2α will be investigated in cervical carcinoma specimens with real-time RT-PCR and western blotting assays. Secondly, it will be ascertained that given viral oncoproteins have correlations to transcription of ASPP1/2 genes and AP-2α with cell model and RNAi technology. Thirdly, it will be comfirmed that ASPP1/2 genes are target ones of transcription factor AP-2αwith chromatin immunoprecipitation, electrophoretic mobility shift,dual luciferase reporter gene system and vectors construction assays. Finally, it will be ascertained that given viral oncoproteins and AP-2α have impact on activity of promoters on ASPP1/2 genes and transcription of ASPP1/2 respectively. Our investigation can provide a new thread and research orientation for further revealing the molecular carcinogenesis mechanism of HPV16 in cervical carcinoma and it is helpful to deepen and richen understanding on carcinogenesis mechanism of HPV16 in cervical carcinoma.

HPV16与宫颈癌发生和发展存在因果关系，病毒癌蛋白与宿主细胞相互作用是研究HPV16致癌分子机制的热点。申请者根据前期研究基础提出科学新观点即HPV16可能介导转录因子AP-2α调控P53凋亡刺激蛋白家族成员ASPP1/2基因的转录是HPV16致宫颈癌新的分子机制。故本项目拟：①核酸扩增和蛋白分析宫颈癌组织中ASPP1/2、AP-2α和HPV16之间的相关性；②RNAi技术明确特定病毒癌蛋白影响ASPP1/2和AP-2α基因转录；③染色质免疫沉淀法、凝胶迁移实验、荧光素酶报告基因系统和DNA重组技术证实ASPP1/2是转录因子AP-2α的靶基因；④RNAi技术证实HPV16特定癌蛋白影响ASPP1/2基因启动子活性以及AP-2α影响ASPP1/2基因的转录水平。本项目为进一步揭示HPV16致宫颈癌分子机制提供新的资料和研究方向,有助于丰富和加深对HPV16致癌分子作用机制的理解和认识。

HPV16与宫颈癌发生和发展存在因果关系，病毒癌蛋白与宿主细胞相互作用是研究HPV16致癌分子机制的热点。申请者根据前期研究基础提出科学新观点即HPV16可能介导转录因子AP-2α调控P53凋亡刺激蛋白家族成员ASPP1/2基因的转录是HPV16致宫颈癌新的分子机制。本项目主要通过IHC、RNA干扰技术、DNA重组技术、核酸扩增技术以及细胞培养等相关实验技术发现，HPV16癌基因不通过转录因子AP-2α下调ASPP1和ASPP2基因的转录即转录因子AP-2α不影响ASPP1和ASPP2基因的转录水平，但是HPV16癌基因下调ASPP1和ASPP2基因的转录并上调转录因子AP-2α基因的转录。本项目为进一步揭示HPV16致宫颈癌分子机制提供新的资料和研究方向,有助于丰富和加深对HPV16致癌分子作用机制的理解和认识。