摘要
目的:探讨胶质瘤干细胞(glioblastoma stem cell,GSC)来源的外泌体(extracellular vesicle,EV)对胶质瘤细胞的增殖、侵袭和替莫唑胺(temozolomide,TMZ)耐药能力的体内外影响。方法:选用原代GSC,通过分化实验获得相对应的分化肿瘤细胞(differentiated glioma cell,DGC)。通过超速离心法分别获得GSC和DGC细胞培养基上清液中的EV。利用CCK8和克隆形成实验、transwell侵袭实验检测GSC来源的EV对胶质瘤U87细胞增殖、TMZ耐药和侵袭能力的影响。小鼠颅内原位成瘤实验分析GSC来源的EV对U87细胞体内增殖、TMZ耐药的影响。结果:通过对原代GSC进行分化,获得了与之相对应的DGC,并且成功获得GSC和DGC来源EV。与DGC来源的EV相比,GSC来源的EV可显著增强U87细胞增殖、侵袭和替莫唑胺耐药能力。体内实验进一步证实,GSC来源的EV能显著促进肿瘤体内生长,明显降低小鼠存活期。结论:GSC来源的EV增强胶质瘤细胞的增殖、侵袭和替莫唑胺耐药能力。
Objective:To investigate the effects of extracellular vesicles(EVs)from glioblastoma stem cells(GSC)on the proliferation,invasion and temozolomide(TMZ)-resistance of glioblastoma(GBM)in vitro and in vivo. Methods:The GSCs were derived from the primary GBM patient and the matched differentiated glioma cells(DGCs)were obtained through differentiation experiments. The EVs in culture medium of GSC and DGC were obtained by ultracentrifugation,respectively. Cell Counting Kit 8(CCK8)assay,colony formation assay and transwell invasion assay were used to detect the effects of GSC-derived EVs on the proliferation,invasion and TMZ-resistance of U87 glioma cells. Intracranial xenografts murine experiment was used to validate the effects of GSC-derived EVs on the in vivo proliferation,invasion and TMZ-resistance of U87 cells. Results:GSC and the matched DGC were obtained through tumor dissociation and differentiation experiments,respectively. GSC and DGC derived EVs were obtained by ultracentrifugation methods. GSC-derived EVs significantly enhanced U87 cell proliferation,invasion and TMZ-resistance,compared to DGC-derived EVs. Consistently,in vivo experiments validated that GSC-derived EVs promoted tumor growth and reduced the survival time of mice. Conclusion:GSC-derived EVs enhance the proliferation,invasion and TMZ-resistance of glioma cells.
作者
高健成
陆晨飞
张梓枫
张军霞
尤永平
GAO Jiancheng;LU Chenfei;ZHANG Zifeng;ZHANG Junxia;YOU Yongping(Department of Neurosurgery,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2022年第12期1658-1663,共6页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金(81974389)。
