miR-150*修饰骨髓间充质干细胞来源的exosome对胶质瘤细胞的影响

miR150*-Loaded exosomes from marrow stromal cells could inhibit glioma cells growth

目的研究miR-150*修饰对骨髓间充质干细胞来源的囊泡(exosome)对胶质瘤细胞的影响。方法 qRT-PCR检测miR-150*在胶质母细胞瘤组织与正常组织间的表达量差异。培养骨髓间充质干细胞(BMSCs),分别转染miR-150*模拟物和阴性对照序列,上调BMSCs中miR-150*表达水平,提取BMSCs培养基中的exosome。Western blot验证exosomal的表面标记蛋白CD63和flotillin-1,电镜下观察exosome的形态。CCK-8和细胞周期实验验证miR-150*修饰BMSCs来源的exosome对胶质瘤细胞的影响。结果 miR-150*在胶质母细胞瘤组织中表达明显低于正常脑组织,转染miR-150*模拟物能明显提高BMSCs来源的exosome中miR-150*的表达。miR-150*修饰BMSCs来源的exosome能有效抑制胶质瘤细胞的增殖。结论 miR-150*(miR-150的互补核苷酸序列)在胶质母细胞瘤组织中低表达,miR-150*修饰BMSCs来源的exosome对胶质瘤细胞有抗增殖的作用。因此exosome可作为一种有效的基因治疗载体。

Objective MiRNA-based therapeutics hold great promise for tumor suppression,this study was to investigate the effect of miR-150＊-loaded exosomes on regulation of glioma cells proliferation and cell cycle. Methods Quantitative real-time PCR on 15 glioblastoma tissues samples and normal controls were used to confirm the miR-150＊expression level. Western blotting analysis and electron microscopy were employed to test exosomal biomarkers and their morphology.Transfection assay were used to collect miR-150＊-loaded exosomes from bone marrow mesenchymal stem cells（ BMSCs） culture medium. CCK-8 and cell cycle assays were used to analyze miR-150＊-loaded exosomes effects on glioma cells. Results Level of miR-150＊expression was much lower in glioblastoma than in normal tissues. Transfection assay successfully acquired miR150＊-loaded exosomes which derived from bone marrow mesenchymal stem cells（ BMSCs）. Furthermore,miR-150＊-loaded exosomes could largely inhibited glioma cells proliferation and suppress cell cycle progression. Cell counting kit 8（ CCK-8） assays also demonstrated miR-150＊delivered in exosomes was much less toxic. Conclusions This study demonstrated miR-150＊is down-regulated in glioblastoma. miR-150＊-loaded exosomes could suppress glioma cells and exosomes may be a potentially efficient therapeutic delivery system.