肿瘤微环境中间充质干细胞相关生物学特性分析.docVIP

肿瘤微环境中间充质干细胞相关生物学特性分析基金项目：国家自然科学基金 基金项目：国家自然科学基金资助项目。 通讯作者： 朱静， Email： HYPERLINK mailto:zhujing310@ zhujing310@。 张亚兰1 朱静1 田杰2 邓兵1 张晓萍1 刘官信1 （重庆医科大学附属儿童医院；1.儿科研究所，重庆市重点实验室2.心血管内科,中国重庆400014） 【摘要】 目的 探讨大鼠骨髓间充质干细胞（bone marrow mesenchymal stem cells,BMSCs）在肿瘤微环境中是否获得肿瘤细胞的相关生物学特性，导致恶性转化。方法 通过六孔板结合transwell小室建立BMSCs和C6胶质瘤细胞的共培养体系,以共培养组为实验组,设单独培养的BMSCs为对照组；相差显微镜下观察培养后两组细胞形态学的改变；流式细胞术(Flow Cytometry,FCM)检测细胞周期变化；荧光定量PCR检测细胞中mdm2、p53 mRNA水平；免疫荧光法检测共培养后p53突变蛋白和mdm2癌蛋白的表达。结果 共培养后实验组细胞表现为胶质瘤细胞样的形态； 实验组G1期细胞比例降低,S期细胞比例升高，无统计学意义(p0.05)；实验组p53 mRNA水平明显降低(p0.05)，部分细胞中表达突变型的p53蛋白,而对照组细胞无突变p53蛋白表达(p0.05)，实验组mdm2 mRNA及其蛋白的表达水平均明显高于对照组(p0.05)。结论 肿瘤微环境使得大鼠骨髓间充质干细胞获得了肿瘤细胞的相关生物学特性,这利用干细胞作为载体治疗肿瘤的安全性研究提供前期实验依据。 【关键词】 骨髓间充质干细胞；肿瘤微环境；共培养；肿瘤转化 The Analysis of Biological Characteristics of Mesenchymal Stem Cells In Tumor Microenvironment ZHANG Ya-lan,ZHU Jing,TIAN Jie,et al（Pediatric Research Institute,Childrens Hospital,Chong Qing Medical University,Chongqing 400014，China） 【Abstract】 Objective: To study whether rat bone marrow-derived mesenchymal stem cells obtain tumor characteristics and have malignant transformation when exposed to tumor microenvironment. Methods: We established co-cultural system between rat bone marrow-derived mesenchymal stem cells and rat glioma cells by using six plate and transwell chamber,cells were divided into two groups:experimental group (co-cultural MSCs),control group (alone cultural MSCs) ;The morphological changes of bone marrow-derived mesenchymal stem cells were observed by phase-contrast microscope;cell cycle were determined by flow cytometry;p53 and mdm2 mRNA were examined by real-time quantitative polymerase chain reaction（PCR）method;the expression of p53 mutant protein and mdm2 protein were detected by immunofluoresce- nce method. Results: Experimental cell show glioma-like cell morphology;cell cycle had no significant changes in experimental group compared with control group(p0.05); p53 mRNA decreased and some cells of experimental