利用“遗传协同致死”模式研究丹参微量弱效多成分的效应倍增机制

Following conducting research of multiple projects supported by the National Natural Science Foundation of China, this group has found that Danshen, which is frequently used in anticancer compounds, could inhibit tumor metastasis and tumor angiogenesis by a special mode different from western medicine (one drug with one target). Domestic scholars put forward that superposition of multicomponent concentrations and synergistic effect caused by multicomponent on multitarget, were the important mechanisms mediated Traditional Chinese Medicine (TCMs) in exerting their pharmacological effects. Our research team will use advease thinking model, and we proposed that synthetic lethality among targets may be the important reason for multiplication effect caused by combination of varieties of traces and weak-potency components. To test this hypothesis, sprouting angiogenesis was selected as the breakthrough of study point. LARS and network topology were employed to build network of tumor sprouting angiogenesis. Then both forward and reverse genetics were utilized, and we found that synchro-suppression of VEGFR-3 and PFKFB could produce the effect of synthetic lethality. On that basis, reverse-phase protein molecular docking technology and the established fluorescent tumor sprout model characterized by a three-dimensional and dynamic manner to conducting biological validation. Furthermore, the cooperative synergism effect of combination of effective components on tumor sprouting, and the effective components of Danshen in inhibition of VEGFR-3 and PFKFB3, respectively, will also be investigated. Successful completion of the proposed studies will not only provide new interpret the mechanism of action of TCM from a new angle, but also provide new thought patterns on research and development of new drug of TCM.

丹参在抗癌方中应用频度很高，课题组在完成多个国家自然基金过程中，证实了丹参可以抑制肿瘤血管生成，且有别于西药一药一靶的作用模式。国内学者提出多成分浓度的叠加及多成分对多靶点的效应协同是中药发挥作用的重要机制。课题组以逆向思维的模式，提出靶标和靶标之间的“遗传协同致死效应”可能是丹参中多种微量、弱效成分组合效应倍增显效的重要原因，为了验证此假说，以肿瘤血管芽生作为研究切入点，结合网络拓扑学和LARS算法构建肿瘤血管芽生网络，利用正/反向遗传学手段，发现同步抑制VEGFR-3和PFKFB3对肿瘤血管芽生具有“协同致死”效应。在此基础上，拟利用反向蛋白分子对接技术结合已建立的三维、可视、动态肿瘤血管芽生模型进行实验验证，研究丹参中分别抑制VEGFR-3和PFKFB3的有效成分，及有效成分组合对抑制肿瘤血管芽生的“协同倍增”效应，从新的角度诠释中药作用机制，也为中药新药研发提供一种全新的思维模式。

丹参在抗癌方中应用频度很高，本研究首先确证了丹参对肿瘤血行转移的抑制作用，且有别于西药一药一靶的作用模式。进一步以肿瘤血管芽生为切入点，以逆向思维的模式，从靶标和靶标之间的“遗传协同致死效应”为策略，结合网络拓扑学和LARS算法构建肿瘤血管芽生网络，筛选出调控血管芽生关键靶标代表性有效成分——隐丹参酮，丹参酮IIA以及丹酚酸B，利用三维、动态、荧光肿瘤血管芽生模型对代表性成分进行了协同机制研究，并进一步基于整体血行转移模型进行了有效成分抑制肿瘤血管生成的贡献性评价。此外，在我们的研究过程中，进一步发现了丹参酮IIA和丹酚酸B进一步促进了肿瘤血管的正常化，重塑了肿瘤内血管的功能，提高化疗药物在肿瘤组织内部的递送。本课题进一步揭示了丹参抑制肿瘤血行转移的科学内涵。