胃癌外泌体激活Shh信号通路诱发MSCs恶性转化及半夏泻心汤干预作用的配伍机制研究

MSCs is the first vector for gastric cancer. Our previous studies show that MSCs can be transformed in gastric cancer microenvironment and can be intervened by Banxia Xiexin Prescription. Its effect is related to its compatibility, but its mechanism has not yet been fully elucidated. The exosomes of gastric cancer can regulate the microenvironment of gastric cancer, induce the proliferation and differentiation of MSCs, and the abnormal activation of Shh pathway can affect the proliferation and differentiation of MSCs. Put forward the work hypothesis of “Banxia Xiexin Prescription regulates Shh signaling pathway to interfere the gastric cancer exosomes induces MSCs malignant transformation”. The aim of this project is to develop non-contact co-culture of exosomes and MSCs in gastric cancer , and observe the expression of MSCs signaling pathway on gastric cancer exocrine body and uptake of key signaling molecules in MSCs Shh pathway, and its growth and proliferation, migration and invasion, and tumorigenesis in vivo. Meanwhile, we will observe the intervention effect of Banxia Xiexin Prescription, and elucidated the effect and mechanism of Banxia Xiexin Prescription on the malignant transformation of gastric cancer induced by exosomatic MSCs, and clarified the relationship between its effect and compatibility. This project from the exocrine pathway of gastric cancer to make a continuous in-depth study of malignant transformation of MSCs in gastric cancer microenvironment It will provide new ideas and new targets for drug action of the development of gastric cancer and the security applications of MSCs to treat gastric cancer

MSCs是胃癌生物治疗的首选载体，但有恶性转化风险。我们前期已证实MSCs在胃癌微环境中可恶性转化并能被半夏泻心汤干预，其效应与该方配伍相关，但其机制尚未完全阐明。胃癌外泌体可调控胃癌微环境、诱发MSCs增殖分化，异常激活的Shh通路可影响MSCs的增殖分化，据此提出“半夏泻心汤调控Shh信号通路干预胃癌外泌体诱发MSCs恶性转化”的工作假说。项目拟将胃癌外泌体与MSCs进行非接触共培养，观察MSCs对胃癌外泌体的摄取及摄取胃癌外泌体后MSCs Shh通路关键信号分子的表达及其生长增殖、迁移侵袭、体内致瘤等生物学特性的变化。同时观察半夏泻心汤的干预作用，阐明半夏泻心汤干预胃癌外泌体诱发MSCs恶性转化的作用及作用机制，明确其效应与配伍的关系。本课题将从胃癌外泌体途径对胃癌微环境中MSCs恶性转化进行连续深入研究，为探讨MSCs的安全应用及胃癌的发生发展提供新思路和药物作用的新靶点。

本研究主要是观察BMSCs对胃癌细胞来源外泌体的摄取及摄取胃癌外泌体后BMSCs的生长、增殖、迁移及侵袭等生物学特性的恶性转化，阐明胃癌细胞来源外泌体诱发BMSCs恶性转化与HH信号通路激活的关系，探讨半夏泻心汤对胃癌细胞来源外泌体诱发BMSCs恶性转化的干预作用及对HH信号通路的影响。通过研究证实胃癌细胞来源外泌体可靶向正常BMSCs并被摄取，半夏泻心汤能够抑制正常BMSCs对胃癌细胞来源外泌体的摄取。胃癌细胞来源外泌体可通过激活HH信号通路诱导BMSCs的生长、增殖、迁移及侵袭，半夏泻心汤可下调胃癌细胞来源外泌体诱导的BMSCs C-myc及HH信号通路中Smo、Shh、Ptch1、Gli1等关键分子表达水平，抑制BMSCs 的生长、增殖、迁移及侵袭。胃癌细胞来源外泌体诱导的BMSCs具有体内致瘤作用，半夏泻心汤可降低LAMP1蛋白，升高TCF-21蛋白的表达，对胃癌细胞来源外泌体诱导BMSCs体内致瘤具有抑制作用，以高剂量作用显著。