脂肪间充质干细胞源exosomes对乳腺癌细胞系MCF7中microRNA表达谱的影响  

The effects of exosomes from adipose-tissue mesenchymal stem cells on the expression of microRNAs in MCF7 cells

作　　者：王世华[1] 林瑞竹[1] 李霄霞[1] 赵春华[1] 

机构地区：[1]中国医学科学院基础医学研究所组织工程中心,北京100005

出　　处：《基础医学与临床》2016年第5期621-626,共6页Basic and Clinical Medicine

基　　金：国家"重大新药创制"国家重大科技专项(2014ZX09101042)

摘　　要：目的观察脂肪间充质干细胞源exosomes的生物学特性，初步探讨其对乳腺癌细胞系MCF7中microRNAs表达谱的影响。方法收集脂肪间充质干细胞培养上清液，以超滤和超离的方法提取exosomes；电镜下观察形态；用Western blot检测exosomes表面蛋白标志物的表达情况；用Dil染料标记exosomes后加入到乳腺癌细胞MCF7上清中，荧光显微镜下观察MCF7对exosomes的内吞作用；用microRNA芯片检测MCF7在exosomes处理24h后microRNAs表达谱的变化以及exosomes中含有的microRNAs种类。结果脂肪间充质干细胞分泌30～100nm的exosomes，exosomes表达CD63和HSP70，可被乳腺癌细胞MCF7内吞；exosomes作用24h，可引起MCF7中244个microRNAs表达发生变化（其中174个表达上调，70个表达下调，倍数〉2倍）；对脂肪间充质干细胞exosomes中microRNAs的分析提示MCF7中microRNAs的上调大部分是内源性的，不是exosomes输入的。结论脂肪间充质干细胞源exosomes可以诱导乳腺癌细胞MCF7microRNA表达谱发生变化，揭示了间充质干细胞影响乳腺癌细胞发生和发展的一个新机制。Objective To evaluate the biological characteristics of exosomes secreted by adipose-tissue derived mesenchymal stem cells（AD-MSCs） and to evaluate the effects of these exosomes on microRNA expression of breast cancer cell MCF7. Methods Exosomes were isolated from supernatant of AD-MSCs by ultracentrifugation and ultrafiltration. The morphology was observed by electron microscopy. Surface marker expression of exosomes was detected by Western blot. Dil-labeled exosomes were added into MCF7 culture medium and the intemalization of exosomes by MCF7 was observed under fluorescence microscope. MicroRNA array chip was used to investigate the microRNA expression changes in MCF7 before and after exosome treatment for 24 h. MicroRNA array chip was also used to detect the expression of microRNAs in exosomes. Results AD-MSCs secreted 30 - 100 nm exosomes which expressed protein makers CD63 and HSP70. Exosomes from AD-MSCs were be internalized by MCF7 and caused microRNA expression changes in MCF7. 244 microRNAs were differentially expressed in MCF7 after treatment with exosomes for 24 h （174 were up-regulated and 70 were down-regulated, with fold change 〉 2 ）. MicroRNA array analysis of exosomes indicated that most of the inereased mieroRNAs in MCF7 were generated endogenously, not derived from exosomes. Conclusions Exosomes from AD-MSCs can regulate microRNA expression of breast cancer cell MCF7, which might be the new mechanism by which AD-MSCs affeet MCF7.

关 键 词：EXOSOMES 间充质干细胞 MICRORNAS MCF7细胞 

分 类 号：R737.9[医药卫生—肿瘤]