外泌体传递miR-139 调控卵巢癌顺铂敏感性的机制研究

目的探讨外泌体介导的miR-139传递对卵巢癌顺铂(cDDP)敏感性的影响。方法分别提取卵巢癌cDDP敏感细胞及耐药细胞分泌的外泌体,透射电镜观察外泌体形态,Western blotting检测外泌体标志蛋白的丰度,qPCR定量检测miR-139水平,MTT检测cDDP敏感细胞、miR-139敲低敏感细胞源性外泌体对耐药细胞cDDP敏感性的影响。结果卵巢癌细胞培养基上清液中存在大量椭圆形或圆形、粒径50~100 nm且高表达外泌体标志因子Alix、CD63、CD9的囊状结构。相较于卵巢癌cDDP耐药细胞源性外泌体,cDDP敏感细胞源性外泌体中miR-139的丰度更高,且将cDDP敏感细胞源性外泌体与耐药细胞株共同孵育可增强后者的cDDP敏感性,而下调外泌体中miR-139的表达可削弱其对耐药细胞的cDDP敏感性诱导作用。结论卵巢癌cDDP敏感细胞可通过外泌体传递miR-139,增强耐药细胞对cDDP的敏感性。

Exosomes Transmit miR-139 to Regulate Cisplatin-Sensitivity in Ovarian Cancer

Objective To investigate the effect of exosome-mediated miR-139 delivery on the regulation of cisplatin(cDDP)sensitivity in ovarian cancer.Methods The exosomes secreted by CDDP-sensitive cells and CDDP-resistant cells were extracted respectively.The morphology of exosomes was observed by transmission electron microscopy.The abundance of exosomes marker proteins was detected by Western blotting,and the expression of miR-139 was detected by qPCR.MTT assay was used to detect the effects of cDDP-sensitive cellderived exosomes and miR-139 knockdown-sensitive cell-derived exosomes on cisplatin sensitivity of drug-resistant cells.Results In the supernatant of ovarian cancer cell culture medium,there were a large number of cystic structures with elliptical or circular shape,particle size of 50~100 nm,and high expression of exosome marker factors Alix,CD63 and CD9.The cDDP-sensitive cell-derived exosomes contained higher abundance of miR-139 than the cDDP-resistant cell-derived exosomes.Moreover,co-incubation of cDDP-sensitive cellderived exosomes with resistant cell lines can enhance the sensitivity of the latter to cDDP,while down-regulation of miR-139 in exosomes can attenuate their induction effects on resistant cells.Conclusion The cDDP-sensitive cells of ovarian cancer can enhance the sensitivity of resistant cells to cDDP by transporting miR-139 via exosomes.