双参颗粒调控S1PR1/STAT3介导的髓源性抑制细胞构筑肺转移前微环境的分子机制

Pre-metastasis niche is a prerequisite for the early stage of tumor metastasis. Recent studies show that myeloid derived suppressor cells (MDSCs) and specific tumor derived factor (TDSFs) play a key role in the formation of lung pre-metastasis niche, and the continuous amplification of MDSCs mediated by S1PR1-STAT3 is a crucial way to promote it. The pathological changes of pre-metastasis niche are correlated with Qi deficiency and blood stasis syndrome, and thus the method of ‘supplementing Qi and activating blood circulation’ is an important rule for prevention of lung metastasis at the early stage. The preliminary studies indicate that the usage of ShuangShen granules guided by this method can inhibit lung metastasis, and reduce the content of MDSCs in lung pre-metastasis niche. On the previous basis, this study used Lewis lung cancer mice marked by GFP+ and co-culture technique for making pre-metastasis niche model, and applied small animal fluorescence imaging system, the high flux of fluorescence enzyme standard instrument, flow cytometry, RT-PCR and other technologies to observe the effect of ShuangShen granules on metastatic outcome through pre-metastasis niche intervention. From MDSCs as the center mediated by S1PR1/STAT3 to set up pre-metastasis niche, this study reveals objective fact and molecular mechanisms of ‘supplementing qi and activating blood circulation’ (ShuangShen granules) to prevent lung metastasis at the early stage, and enrich the Chinese medical theory of ‘preventive treatment of disease’.

转移前微环境形成是肿瘤早期发生转移的先决条件。研究表明：髓源性抑制细胞（MDSCs）与特异性肿瘤源性分泌因子在肺转移前微环境形成中具有关键调控作用，而S1PR1/STAT3介导的MDSCs持续扩增是促进其形成的重要途径。肺转移前微环境的病理改变符合气虚血瘀证的生物学特征，益气活血法是早期预防肺转移的重要法则，前期研究显示：其治则下的双参颗粒具有抑制肺转移的作用，并可明显降低肺转移前微环境中MDSCs含量。本课题在前期基础上，利用GFP+标记的Lewis肺癌小鼠与共培养技术建立肺转移前微环境模型，运用小动物荧光成像、荧光酶标仪高通量精准检测、FACS、RT-PCR等技术，动态观察双参颗粒干预转移前微环境对转移结局的影响，从S1PR1/STAT3介导的MDSCs为中心构筑肺转移前微环境的新视角，揭示益气活血法早期预防肺转移的分子机制及客观规律，丰富中医肿瘤“治未病”理论。

近年研究表明转移前微环境形成是肿瘤早期发生转移的先决条件，髓源性抑制细胞（MDSCs）与特异性肿瘤源性分泌因子在肺转移前微环境形成中具有关键的调控作用，其中S1PR1/STAT3是促进转移前微环境中MDSCs持续扩增的重要信号通路。益气活血法是早期预防肺转移的重要法则，本课题在前期基础上，利用GFP+标记的Lewis肺癌小鼠与共培养技术建立体内外转移前微环境模型，运用小动物荧光成像、荧光酶标仪高通量精准检测、FACS、RT-PCR等技术，从S1PR1/STAT3介导的MDSCs为中心构筑肺转移前微环境的新视角进行研究，研究结果表明：（1）双参颗粒对原位肿瘤有一定的抑制作用，并可在一定程度上减少肺转移灶。（2）双参颗粒可有效减少肺转移前微环境中MDSCs的含量，可能与降低肿瘤细胞分泌的部分TDSFs有关。（3）肿瘤细胞s1pr1-stat3信号通路的激活与骨髓细胞向MDSCs的转化、肺脏s1pr1-stat3激活有关。双参颗粒可能通过降低肿瘤细胞s1pr1-stat3信号通路的激活程度，抑制骨髓细胞向MDSCs的转化，减少MDSCs在肺脏转移前微环境中的聚集。（4）双参颗粒通过降低肺转移前微环境中s1pr1-stat3信号通路的激活程度及MDSCs的积累，减少转移前微环境标记物Version、Fibronectin、Lox、MMP9的表达水平，逆转肺转移前微环境，减少肿瘤细胞肺转移。课题揭示益气活血法（双参颗粒）早期预防肺转移的分子网络机制，丰富中医肿瘤“治未病”理论。