基于外泌体中ceRNA网络表达研究温肾散结法干预前列腺癌骨转移免疫微环境的作用机制研究

The most common metastatic mode of Prostate cancer is bone metastasis and we had little clinical treatment to treat bone metastasis of prostatic cancer. Base on the pathogenesis which is deficiency of kidney-yang and stagnation of yin-cold, it can decrease the PSA level in patients and improve their life qualities that use the method of warming the kidney and eliminating stagnation. By previous experiments, we discovered the method of warming the kidney and eliminating stagnation can inhibit the prostate cancer cell proliferation through P13K/AKT and mTOR signaling pathway. Further data analysis also found that the metastasis of prostate cancer was related to regulatory network of ceRNA and non-coding genes of immune pathways in exosome. Therefore, we hypothesized thatthe method of warming the kidney and eliminating stagnation can treat bone metastasis of prostatic cancer by regulating the gene network in exosome to change immune microenvironment of bone metastasis of prostatic cancer. The experiment from two aspects of tumor tissues and exosome will research the specific response between the circRNA and miRNA during the treatment and explore the mechanism of action that Chinese Herbs treats bone metastasis of prostatic cance by affecting the ceRNA network and P13K/AKT signaling pathway to change immune microenvironment of bone metastasis. Subsequently, we will test the mechanism of action of the method of warming the kidney and eliminating stagnation by nude mice model and function tests and provide scientific basis.

前列腺癌最易发生骨转移，临床无有效治疗手段。课题组基于晚期前列腺癌骨转移“肾阳衰败、阴寒凝结”的病机特点，运用“温肾散结法”能有效降低患者前列腺抗原PSA水平、改善生存质量。我们的前期实验研究也发现，温肾散结法通过PI3K/AKT及mTOR通路抑制前列腺癌细胞增殖。进一步数据挖掘，发现本病的转移与外泌体中ceRNA调控网络及免疫通路非编码基因相关。故我们提出假说：温肾散结法通过调控外泌体中ceRNA网络表达干预前列腺癌骨转移免疫微环境，并治疗前列腺癌骨转移。本研究拟通过基因芯片技术，从“肿瘤组织”与“血液外泌体”两方面，研究“温肾散结法”治疗过程中特异性响应circRNA与miRNA，挖掘中药影响ceRNA网络表达调控PI3K/AKT信号通路改变骨转移免疫微环境治疗前列腺癌骨转移的作用机制。最后通过裸鼠模型、功能实验验证“温肾散结法”的作用机制。为温肾散结法治疗前列腺癌骨转移提供科学依据。

前列腺癌是常见的泌尿系统恶性肿瘤之一，世界范围内，前列腺癌发病率在男性所有恶性肿瘤中位居第二。骨骼是最常见的前列腺癌转移靶器官，超过80%的前列腺癌患者会发生骨转移，针对晚期骨转移患者，众多内分泌和放化疗方法的临床疗效均不理想且费用高昂，迫切需要新的药物研发支持。课题组基于前期研究提出假说，认为温肾散结法能够通过调控外泌体中ceRNA网络表达干预前列腺癌骨转移免疫微环境，从而发挥治疗前列腺癌骨转移的作用。.本项目筛选并验证了前列腺癌组织与血浆外泌体两个层面上，与骨转移和用药相关的转录组水平表达差异，构建了相关ceRNA互作网络，相关实验结果证明了温肾散结中药可以通过干预“circ RNA_0070938 - miR-218 - BCAT1”ceRNA调控网络发挥干预前列腺癌骨转移的作用。通过检测前列腺癌骨转移患者用药前后血浆中成骨分化标志物和破骨活性标志物等细胞因子的变化，发现温肾散结组方对成骨分化具有促进作用，而对破骨分化呈现抑制作用。并通过检测模型裸鼠血样PI3K/AKT 通路标记物（PI3K、p-AKT）水平及特异性细胞因子（白介素IL-6、IL-10）水平的表达，揭示中药可能通过调控PI3K/AKT通路及抑制IL-6、IL-10因子表达改善不良免疫微环境从而激活免疫细胞活性干预前列腺癌骨转移。.综上，本项目在前期研究基础上进一步从外泌体ceRNA调控网络角度进行研究，解决了部分科学问题，包括前列腺癌骨转移过程中关键ceRNA调控网络的组成及差异，以及温肾散结法如何通过影响ceRNA网络和细胞因子调控免疫微环境干预前列腺癌骨转移。本项目运用以上研究模式，为中医药治疗前列腺癌骨转移作用的分子机制，探索出一条新的研究模式，为温肾散结法及相关中药组方的科学应用和发展打下了基础。