辐射诱导的p73变异体对肿瘤细胞凋亡的调控作用

This project focused on the tumor hotspot of p73 gene, which has a double-sided function of oncogene and tumor suppressor gene. This project explored the role of radiation-induced p73 variants in tumor cell apoptosis, and functional differences between the different variants. The proposed model of tumor cell apoptosis induced by ionizing radiation to detect differentially expressed between the different p73 variants through gene and protein expression; and p73 variants were screened by lipofection radiation sensitivity of different tumor cell lines, using flow cytometry and TUNEL to measure the impact of the variants of the cell cycle and apoptosis; Through gene expression, immunocytochemistry and laser scanning confocal, interaction between different p73 variants was detected. The aim of this project is to explore the radiation-induced tumor cell apoptosis molecular mechanism regulated by different p73 variants. This study is an important supplement to the apoptosis mechanism caused by the radiation, the p73 gene as a target for the design of radiation therapy sensitizer technology, to further improve the efficacy of the carcinogenicity of heavy ions to the theoretical basis.

聚焦于p73基因具有癌基因和抑癌基因的双面功能这一肿瘤研究热点。本项目探索辐射诱导的p73变异体在肿瘤细胞凋亡中所起的作用，及其不同变异体之间的功能差异。拟采用离子辐射诱导的肿瘤细胞凋亡模型，通过基因和蛋白表达等方法检测不同p73变异体之间的差异表达；将筛选得到的p73变异体通过脂质体转染辐射敏感性不同的肿瘤细胞株，运用流式细胞仪和TUNEL法测量该变异体对细胞周期和凋亡的影响；通过基因表达、免疫细胞化学、激光共聚焦法等检测不同p73变异体之间的相互作用。拟证实辐射诱导不同p73变异体调控肿瘤细胞凋亡分子机制。此研究是对辐射造成细胞凋亡机理的重要补充，为设计以p73基因为靶点放射治疗增敏技术，进一步提高重离子致癌疗效奠定理论基础。

重离子治癌技术值得信赖。但重离子治癌技术临床应用时间较短，其基础和临床研究工作还有待加强,尤其是重离子杀伤肿瘤细胞机理的研究有待深入研究。因此，本项目聚焦于p73基因具有癌基因和抑癌基因的双面功能这一肿瘤研究热点，研究了辐射诱导的凋亡“开关”p73对肿瘤细胞凋亡的调控机制，探索了辐射诱导的p73变异体在肿瘤细胞凋亡中所起的作用，及其不同变异体之间的功能差异。该研究采用了离子辐射诱导的肿瘤细胞凋亡模型，通过荧光定量PCR、Western blot、免疫组化、免疫荧光等方法检测了不同p73变异体之间的差异表达；运用流式细胞仪和TUNEL法测量该变异体对细胞周期和凋亡的影响；通过基因表达、免疫细胞化学、激光共聚焦法等检测不同p73变异体之间的相互作用。该项目建立了射诱导的细胞凋亡模型，证实了辐射诱导的TAp73和DNp73动态平衡在肿瘤细胞凋亡中所起的关键作用，阐明了提高TAp73和DNp73的比例可以提高肿瘤细胞的辐射敏感性，深入了解基因的选择性剪接造成的基因复杂度，蛋白和功能的多样性。总之，此研究是对辐射造成细胞凋亡机理的重要补充，为设计以p73基因为靶点放射治疗增敏技术，进一步提高重离子致癌疗效奠定理论基础。