人脐带间充质干细胞来源的微囊通过诱导HGF促进人肾癌细胞的生长和迁移侵袭能力

发布时间：2018-07-05 13:38

本文选题：人脐带间充质干细胞 + 微囊　；参考：《上海交通大学》2014年硕士论文

【摘要】：目的：初步探讨人脐带间充质干细胞来源的微囊（hWJMSC-MVs）对人肾癌细胞(RCC）的影响及其可能的作用机制。方法：1、选用人肾癌细胞株786-O作为研究对象，分三组进行干预：（1）+MVs组，（2）control组，（3）+RNase-MV组（经RNA酶处理的MV），24h、48h后检测肿瘤细胞增殖率（CCK-8）及细胞周期，进行划痕实验检测。运用实时定量PCR检测肾癌细胞中MMP-2、MMP-9基因的表达情况。 2、裸鼠皮下注射肾癌细胞建立肾癌移植瘤模型，每三天测量一次肿瘤大小，计算各组肿瘤面积及发生率，，36d后进行牺牲，分别进行免疫组化分析（HGF、KI-67及Tunel）。 3、运用Western blot技术检测各组肾癌标本中MMP-2、MMP-9、cyclinD1的蛋白表达情况。然后检测各组肾癌细胞或肿瘤标本中HGF、c-MET、ERK1/2、AKT蛋白表达情况，并对c-MET抑制剂干预后的肾癌细胞中p-ERK1/2、p-AKT蛋白表达情况进行检测。结论：hWJMSC-MVs促进了肾癌的生长及侵袭能力，其作用机制可能是MVs诱导HGF的增加从而增强了PI3k-AKT及Ras/MAPK（ERK1/2）信号通路的活化，进一步加强了肾癌细胞增殖及抗凋亡的能力。同时发现MV经RNA酶处理后丧失了进一步促进肾癌生长的能力，证明MVs对靶细胞的作用可能是通过传递有效的遗传物质（mRNA/miRNA）来实现的，MVs可能是介导细胞间相互作用的一种媒介物质。本实验也为HGF/c-MET途径可能成为肾癌治疗的靶点提供了实验依据。
[Abstract]:Aim: to investigate the effect of human umbilical cord mesenchymal stem cell derived microcapsules (hWJMSC-MVs) on human renal cell carcinoma (RCC) and its possible mechanism. Methods: 1. Human renal cell carcinoma cell line 786-O was selected as the study object. It was divided into three groups: (1) MVs group, (2) control group, (3) RNase-MV group (RNase-MV) 24 h and 48 h later, the tumor cell proliferation rate (CCK-8) and cell cycle were detected by scratch test. The expression of MMP-2 and MMP-9 gene in renal carcinoma cells was detected by real-time quantitative PCR. 2. The transplanted tumor model was established by subcutaneous injection of renal cancer cells in nude mice, and the tumor size was measured every three days. After 36 days' sacrifice, the tumor area and incidence of each group were analyzed by immunohistochemical analysis (HGFKI-67 and Tunel). Western blot was used to detect the expression of MMP-2, MMP-9 and cyclin D1 in renal cell carcinoma. Then, the expression of HGF- c-METERK1 / 2AK protein in renal cancer cells or tumor samples was detected, and the expression of p-ERK1 / 2 pAKT protein in renal cancer cells after intervention with c-MET inhibitor was detected. Conclusion the growth and invasiveness of RCC were promoted by WJMSC-MVs. The mechanism may be that MVs induces the increase of HGF, which enhances the activation of PI3k-AKT and Rasr-MAPK (ERK1 / 2) signaling pathway, and further strengthens the proliferation and anti-apoptosis ability of RCC cells. At the same time, it was found that MV lost the ability to further promote the growth of renal cell carcinoma after being treated with RNAase, which suggested that MVs' effect on target cells might be mediated by the transmission of effective genetic material (mRNA-miRNA). This study also provides experimental evidence that HGF-c-MET pathway may be a target for the treatment of renal cell carcinoma.
【学位授予单位】：上海交通大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.11

【共引文献】