损伤来源外泌体促进胶质母细胞瘤细胞化疗抗性

目的观察细胞机械损伤后的外泌体对细胞化疗药物的影响，初步探讨手术造成的损伤对胶质母细胞瘤（GBM）细胞化疗抗性的影响。方法用含10%胎牛血清的DMEM完全培养基培养人GBM细胞U251及U87，分为对照组与损伤组。更换无血清培养基，损伤组使用10 μL枪头在贴壁细胞层上快速划痕后继续培养12 h；对照组同步换液培养，收集上清并与新鲜培养液等比例培养细胞，CCK-8、克隆形成、EdU实验检测细胞增殖情况，差速离心法分离上清中的外泌体，用等量的外泌体（75 μg/mL）培养细胞，通过CCK-8、克隆形成和EdU实验检测外泌体对细胞增殖的影响，IC50实验检测细胞对替莫唑胺与多柔比星的敏感性。结果损伤上清可促进细胞的增殖，其外泌体不仅显著促进细胞增殖能力，也显著促进细胞对化疗药物的耐受性。结论细胞受到机械损伤后分泌的外泌体促进GBM细胞的化疗抗性。

Objective To observe the effect of exosomes on chemotherapeutic drugs after cell mechanical injury, and to explore the effect of surgical injury on chemoresistance of glioblastoma(GBM) cells. Methods The human GBM cells U251 and U87 were cultured in DMEM complete medium containing 10% fetal bovine serum, and divided into the control group and the injury group. The serum-free medium was replaced. The injury group was quickly scratched on the adherent cell layer with a 10 μL pipette tip and then cells were cultured for additional 12 hours. The control group was synchronously changed for culture, and the supernatant was collected and cultured in the same proportion as the fresh culture medium. The CCK-8,clone formation and EdU experiments were conducted to detect cell proliferation. The exosomes in the supernatant were separated by differential centrifugation, and cells were cultured with an equal amount of exosomes(75 μg/mL). The effects of exosomes on cell proliferation were detected by CCK-8, clone formation and EdU experiments, and the sensitivity of cells to temozolomide and doxorubicin was detected by IC50 experiment. Results The injury supernatant could promote cell proliferation, and the exosomes not only significantly promote cell proliferation, but also significantly promote cell tolerance to chemotherapeutic drugs. Conclusion Exosomes secreted by cells after mechanical damage promote chemotherapy resistance in GBM cells.