基于计算机拟合的类雄激素的筛选及其介导雄激素非依赖性前列腺癌发生发展的机制研究

Androgen-independent prostate cancer (AIPC) is not only the leading cause of death in patients with prostate cancer, but also a difficult issue for current treatment. According to the latest theory, PCa recurrence after castration is not really "androgen-dependent", on the contrary, the reactivation of the androgen receptor (AR) signaling pathway was considered to be an important mechanism for the development of AIPC. Studies showed that high cholesterol levels and cancer development were closely linked, and testosterone was generated by the conversion of cholesterol in vivo. This work is based on the concept of translational medicine with the assistance of computer virtual screening technology, through studying the combination force of cholesterol metabolites in vivo and the androgen receptor, looking for possible endogenous substances induced hormone-independent prostate cancer, using gene knock and plasmid construction techniques to investigate the effects of candidate compounds for PCa from in vitro in vivo, clarify the relationship of cholesterol metabolism and development of PCa, reveal underlying mechanisms of AR downstream signaling （PSA,KLK2,FKBP5,HOMER2,TRPMH） with cholesterol metabolites, in order to provide theoretical basis and potential treatment options for prostate cancer treatment.

雄激素非依赖性前列腺癌（AIPC）是前列腺癌（PCa）患者死亡的主要原因，也是目前治疗的难点。最新研究发现，去势后复发并非真的“激素非依赖”，相反，雄激素受体（AR）信号通路的再激活被认为是AIPC发生发展的一个重要机制，然而其激活机制未明。研究发现由高胆固醇引起的代谢综合症与癌症的发生发展有密切的联系，胆固醇在体内有复杂的代谢产物，雄激素在体内也是由胆固醇转化而来。本课题基于转化医学的理念，应用计算机虚拟筛选技术，通过研究胆固醇代谢物与雄激素受体结合力的分析，寻找可能诱发AIPC的内源性物质，应用基因敲除、质粒构建等技术，分别从分子、细胞和动物三个水平考察候选代谢物对AIPC的影响，阐明胆固醇代谢产物与PCa发生发展的关系，揭示胆固醇代谢产物与PCa去势后AR再激活下游信号分子（PSA,KLK2,FKBP5,HOMER2,TRPMH）的内在机制，为研发新的治疗药物提供理论基础和实验依据。

雄激素非依赖性前列腺癌（AIPC）是前列腺癌（PCa）患者死亡的主要原因，也是目前治疗的难点。最新研究发现，雄激素受体（AR）信号通路的再激活被认为是AIPC发生发展的一个重要机制，然而其激活机制未明。本研究通过计算机虚拟筛选技术发现孕酮与雄激素受体结合力高，临床样本分析显示孕酮的表达与前列腺癌患者体内雄激素受体负相关。进一步研究显示孕酮可以明显抑制AR敏感前列腺癌细胞(LNCaP)的增殖且细胞活力明显降低，优于临床用药恩杂鲁胺，此外同等剂量下孕酮的细胞毒性作用低于恩杂鲁胺。对于AR不敏感前列腺癌细胞（22RV1）,孕酮与恩杂鲁胺相比，细胞毒性明显较低但IC50相当。在22RV 1和MYC-Cap小鼠前列腺癌细胞中，我们通过CI指数，探索出孕酮与恩杂鲁胺的联合用药剂量比，细胞增殖曲线及克隆形成试验结果均显示，两者的此种比例联用效果最优。随后我们构建了AR靶基因双荧光素酶报告质粒pMMTVv-Luc，结果显示孕酮与恩杂鲁胺联用可以有效抑制雄激素受体对AR靶基因的转录功能。