益气补肾方调控胃癌干细胞niche及Notch信号通路抗转移分子机制研究  

Study on Molecular Mechanisms of Anti-proliferation and Metastasis of Yiqi Bushen Formula by Regulating the Gastric Cancer Stem Cell Niche and Notch Signaling Pathway

作　　者：刘云霞[1] 徐叶峰[1] 蒋沈君[1] 姚勇伟[1] 王娜[2] 

机构地区：[1]杭州市第三人民医院,杭州310009 [2]浙江中医药大学第二临床医学院

出　　处：《浙江中医药大学学报》2016年第7期514-522,共9页Journal of Zhejiang Chinese Medical University

基　　金：国家自然科学基金面上项目(81473505);浙江省中医药科技计划项目(2013ZB105);杭州市科技局项目(20130733Q23)~~

摘　　要：[目的]探讨益气补肾方调控胃癌肿瘤干细胞(cancer stem cells,CSC)niche及Notch信号通路抗胃癌增殖转移的分子机制,为临床应用提供实验依据。[方法]采用人胃癌细胞株NCI-N87建立胃癌CSC富集裸鼠荷瘤模型。裸鼠随机分为阴性对照组、模型对照组、氟尿嘧啶组(5-Fu组)、益气补肾方组(益气补肾组)及益气补肾方+氟尿嘧啶组(协同组)。每组药物干预6周,观察裸鼠体质量变化及瘤体生长情况,比较各组肿瘤增殖及远处转移情况;采用ELISA法检测瘤体血管内皮生长因子受体-1(vascular endothelial growth factor receptor-1,VEGFR-1)、骨桥蛋白(osteopontin,OPN)、缺氧诱导因子-1α(hypoxia-inducible factor-1α,HIF-1α)、CXC趋化因子受体4(CXC chemokine receptor 4,CXCR4)表达;采用免疫组化法检测肿瘤组织中CD24、CD44、上皮细胞粘附分子(epithelial cell adhesion molecule,Ep CAM)的表达;采用Western blot检测肿瘤组织Notch1、Jagged1蛋白表达。[结果]给药6周后,益气补肾组、协同组及阴性对照组裸鼠体质量均明显重于模型对照组、5-Fu组,差异均有统计学意义(P<0.01);模型对照组与阴性对照组裸鼠的瘤体均重于5-Fu组、益气补肾组及协同组,差异有统计学意义(P<0.01)。5-Fu组、益气补肾组及协同组裸鼠的肝脏、腹壁、淋巴结及肺的总转移率明显少于模型对照组,差异有统计学意义(P<0.05)。与模型对照组比,5-Fu组、益气补肾组及协同组瘤体内VEGFR-1、OPN、HIF-1α、CXCR4的表达下调,差异有统计学意义(P<0.05,P<0.01)。益气补肾组与协同组CD24、CD44的表达均低于模型对照组及5-Fu组,差异有统计学意义(P<0.05)。益气补肾组Ep CAM的表达低于模型对照组及5-Fu组,差异有统计学意义(P<0.05)。益气补肾组及协同组瘤体组织中Notch1、Jagged1蛋白表达均低于模型对照组,差异有统计学意义(P<0.05)。[结论]益气补肾方具有显著的抗胃癌细胞增殖转移的作用,其机制可能[Objective]To investigate the molecular mechanisms of anti-proliferation and metastasis of Yiqi Bushen Formula by regulating the gastric cancer stem cell niche and Notch signaling pathway, in order to provide the theory basis for its clinical application. [Methods]The nude mice of human gastric carcinoma constructed using orthotopic transplantation and an enrichment in cancer stem cell of NCI-N87 gastric cancer cell line were randomized into negative control group, model control group, 5-fluorouracil group, Yiqi Bushen Formula group,combination group of Yiqi bushen Formula and 5-fluorouracil（collaborative group）. After being treated with different drugs for 6 weeks, observe and analyze changes in nude mice body weight, the growth and change of the transplanted tumor, metastasis rate and inhibition rate of tumor. Detecting the expression quantity in tumor tissues of vascular endothelial growth factor receptor-1（VEGFR-1）, osteopontin（OPN）, hypoxia-inducible factor-1α（HIF-1α）, CXC chemokine receptor 4（CXCR4） by Elisa method.Investigating the expression of CD24, CD44, epithelial cell adhesion molecule（Ep CAM） in tumor tissues by immunohistochemical method. Detecting the expression level of Notch1, Jagged1 in different tumor tissues by Western blot. [Results] The average weight of Yiqi Bushen Formula group, collaborative group and negative control group became significantly higher than those in model control group and 5-fluorouracil group（P〈0.01）. Meanwhile, the weight of the transplanted tumor in model control group and negative control group was significantly higher than those three treatment groups（P〈0.01）. The incidence rates of metastasis in liver,abdominal wall,lymph nodes and lung in 5-Fu group, Yiqi Bushen Formula group and collaborative group were significantly lower than the other two control groups（P〈0.05）. Compared with model control group, the expression level of VEGFR-1, OPN, HIF-1α, CXCR4 significantly decreased in treatment groups（P〈0.05）. The expressi

关 键 词：益气补肾方 胃癌 CSC NICHE NOTCH信号通路 增殖转移 分子机制 

分 类 号：R331[医药卫生—人体生理学]