人脐带间充质干细胞外泌体递送外源miR-130a-3p抑制人肺癌A549细胞的研究

Exosomes derived from human umbilical cord mesenchymal stem cells deliver exogenous miR-130a-3p to inhibit human lung cancer A549 cells

目的观察人脐带间充质干细胞(hUC-MSCs)外泌体介导的外源miR-130a-3p摄取对肺癌A549细胞的影响。方法采用超速离心法分离hUC-MSCs外泌体,透射电子显微镜鉴定外泌体形态和大小,Westernblot检测外泌体标志性蛋白(CD63、CD81)表达,纳米粒度分析仪(NTA)测定外泌体颗粒浓度和粒径分布;通过瞬时转染制备负载miR-130a-3p mimic的hUC-MSCs外泌体(130a-exo),用130a-exo处理A549细胞,RT-qPCR检测A549胞内miR-130a-3p含量,CCK-8法测定细胞增殖,Transwell实验检测细胞迁移,流式细胞术检测细胞凋亡。结果从hUC-MSCs培养上清中分离的纳米颗粒符合外泌体的典型特征。RT-qPCR结果显示,与对照组相比,130a-exo组A549细胞的miR-130a-3p水平显著升高(P<0.01)。与对照组相比,130a-exo组A549细胞的增殖能力显著降低(P<0.01),体外迁移能力明显下降(P<0.05),早期凋亡率显著升高(P<0.05),晚期凋亡率亦明显增加(P<0.01)。结论hUC-MSCs外泌体可将外源miR-130a-3p递送至肺癌A549细胞而发挥抑癌作用。

Objective To observe the effects of exogenous miR-130a-3p delivered by exosomes derived from human umbilical cord mesenchymal stem cells(hUC-MSCs)on lung cancer A549 cells.Methods Exosomes were isolated from culture supernatant of hUC-MSCs using the ultracentrifugation method.The shape and size of exosomes were identified by transmission electron microscopy,and the expression of marker proteins of exosomes was detected by western blot.The particle concentration and the size distribution of exosomes were measured by nanoparticle tracking analysis(NTA).The miR-130a-3p mimic was loaded into hUC-MSC exosomes(130a-exo)by transient transfection,which was then used to treat A549 cells.The miR-130a-3p level in A549 cells was examined by RT-qPCR.Cell proliferation,migration and apoptosis of A549 cells treated with 130a-exo were detected by CCK-8 method,Transwell experiment and flow cytometry,respectively.Results The nanoparticles derived from hUC-MSCs conformed to the typical characteristics of exosomes.Compared with the control group,the miR-130a-3p level was significantly elevated in A549 cells of 130a-exo group(P<0.01).Exogenous miR-130a 3p obviously inhibited cell growth(P<0.01)and migration(P<0.05)of A549 cells.Meantime,130a-exo induced the apoptosis of A549 cells evidenced by the increasing of both early apoptosis rate(P<0.05)and late apoptosis rate(P<0.01).Conclusion Exosomes derived from hUC-MSCs can deliver exogenous miR-130a-3p to lung cancer A549 cells and play an anti-cancer role.