益气补肾方调控胃癌干细胞免疫微环境及Notch信号通路抗转移分子机制研究

Gastric cancer is a common malignant tumor, of which five-year survival rate is no more than 20 percent, and above 50 percent died of recurrence and metastasis. Cancer stem cells (CSC) are cells of the small part of the malignant tumor tissue with strong ability of self-renewal and undirectional differentiation, consequently the transfer and homing of CSC is the primary cause of tumor metastasis. Studies confirm that the formation of immune suppressive microenvironment in CSC niche provides the initial power for the start of transfer and homing,while the activation of Notch signaling pathway and the crosstalk in regulatory netwoks accelerating this progress.Therefore,the therapy targeting immune microenvironment in CSC niche and Notch signaling pathway may provide new strategies on anti-metastasis treatment. The preliminary study of the project team found that Yiqi Bushen Formula has significant effect of anti-metastasis on gastric cancer.This project will observe the influence on each link of proliferation,differentiation and metastasis in gastric CSC by Yiqi Bushen Formula,and through which we may clarify the molecular mechanisms of anti-metastasis of Yiqi Bushen Formula by regulating the immune suppressive microenvironment and Notch signaling pathway,and finally provide the theory basis for its clinical application in preventing recurrence and metastasis of gastric cancer.

胃癌是最常见恶性肿瘤之一，五年生存率不超过20%，超过50%死于复发和转移。肿瘤干细胞(CSC)是恶性肿瘤组织中一小部分具有极强自我更新能力和不定向分化能力的细胞，CSC的转移和归巢是肿瘤复发转移的根本原因。研究证实，CSC小生境（CSC niche）中免疫抑制微环境的形成为CSC转移的启动和归巢提供初始动力，而Notch信号通路的活化及其在细胞多维调控网络中的串话加速了CSC的转移和归巢行为。因此，靶向CSC niche免疫微环境和Notch信号通路的治疗将会给抗肿瘤转移治疗提供新策略。项目组前期临床和实验研究已发现益气补肾方具有较好的抗胃癌转移作用，本课题将进一步通过观察益气补肾方对胃癌CSC增殖、分化及转移各个环节的影响，明确其整体调控胃癌CSC niche免疫微环境和Notch信号通路抗胃癌转移的分子机制，为中医药防治胃癌复发及转移的临床应用提供新的理论依据。

胃癌是最常见的消化道恶性肿瘤之一，五年生存率不超过20%，超过50%死于复发和转移。肿瘤干细胞(CSC)是恶性肿瘤组织中一小部分具有极强自我更新能力和不定向分化能力的细胞，CSC的转移和归巢是肿瘤复发转移的根本原因。CSC小生境（CSC niche）中免疫抑制微环境的形成为CSC转移的启动和归巢提供初始动力，而Notch信号通路的活化及其在细胞多维调控网络中的串话加速了CSC的转移和归巢行为。因此，靶向CSC niche免疫微环境和Notch信号通路的治疗将会给抗胃癌转移治疗提供新策略。本项目通过体内外实验，观察益气补肾方对胃癌CSC增殖、分化及转移各个环节的影响，阐明益气补肾方多层次、多靶点抗胃癌转移的分子机制。体外实验研究发现，CD44、EpCAM可作为胃癌SGC-7901细胞CSC表面标志物；随着给药浓度的增加和给药时间的延长，益气补肾方含药血清能显著抑制SGC-7901增殖生长；益气补肾方干预后各组细胞中CD44+EpCAM+干细胞比例下调；益气补肾方能明显改变胃癌CSC的细胞形态和内在超微结构；益气补肾方能显著抑制SGC-7901侵袭转移。体内实验表明，益气补肾方能显著抑制胃癌原位移植瘤的生长和转移，能降低胃癌CSC表面标志物CD44、EpCAM的表达，降低血清TGF-β1表达水平。益气补肾方干预后，瘤体内肿瘤转移相关因子TGF-β1、VEGFR-1、OPN、HIF-1α、CXCR4的表达均有降低，Notch1、Jagged1蛋白及mRNA表达均下调。本研究明确了益气补肾方整体调控胃癌CSC niche免疫微环境和Notch信号通路抗胃癌转移的分子机制，为中医药防治胃癌复发及转移的临床应用提供新的理论依据。项目资助发表核心论文6篇，申请国家发明专利1项，参加国内外学术会议交流4次，获得杭州市科技进步奖1项，培养硕士研究生3名。