摘要
目的:探讨外泌体(exosome,Exo)来源的miR-145传递对上皮性卵巢癌SKOV3细胞增殖、迁移、侵袭、血管形成及癌干细胞样特性的影响。方法:卵巢癌SKOV3细胞转染miR-145 mimic、miR-145 inhibitor或miR-145阴性对照(NC)。分离并鉴定SKOV3细胞分泌的Exo和miR-145过表达/低表达的SKOV3细胞分泌的外泌体。采用RT-qPCR法检测miR-145的表达。SKOV3细胞与不同来源的外泌体共培养。MTT法和Transwell法检测细胞增殖、迁移和侵袭。血管形成实验检测外泌体miR-145对人脐静脉内皮细胞血管生成能力的影响。采用肿瘤球形成实验评估SKOV3细胞系中CD133^(+)/CD44^(+)细胞亚群的干细胞特征。结果:miR-145 mimic-Exo处理后SKOV3细胞中miR-145表达显著上调,miR-145 inhibitor-Exo处理后SKOV3细胞中miR-145表达明显下调(P<0.05)。卵巢癌细胞来源的Exo可促进SKOV3细胞增殖、迁移和侵袭(P<0.05)。高水平的外泌体miR-145可降低SKOV3细胞增殖、迁移和增殖,抑制人脐静脉内皮细胞的血管生成,抑制SKVO3细胞形成肿瘤球的能力(P<0.05)。然而,miR-145表达下调的外泌体具有相反的作用(P<0.05)。结论:通过外泌体传递miR-145能够抑制上皮性卵巢癌细胞迁移、侵袭以及血管生成,并调控上皮性卵巢癌干细胞样特征。
Objective:To investigate the effects of epithelial ovarian cancer(EOC)cell-secreted exosomal miR-145 on the proliferation,migration and invasion abilities and stem celllike characteristics of EOC cells.Methods:Ovarian cancer SKOV3 cells were transfected with miR-145 mimic,miR-145inhibitor,or miR-145 negative control(NC).Exosomes(Exo)derived from SKOV3 cells and miR-145-overexpressing or miR-145-downregulated SKOV3 cells were isolated and identified.The expression of miR-145 was assessed by RT-qPCR.SKOV3cells were co-cultured with exosomes from different sources.Cell proliferation,migration and invasion were detected by MTT assay and Transwell assay.The impact of exosomal miR-145on angiogenic tube formation ability of endothelial cells was tested by tube formation assay.The stem cell characteristics of CD133^(+)/CD44^(+)cells sorted from SKOV3 cell line were evaluated by tumor sphere formation assay.Results:miR-145 expression was significantly upregulated in SKOV3 cells treated with miR-145 mimic-Exo and downregulated in cells treated with miR-145 inhibitor-Exo.Exo derived from ovarian cancer cells favored the proliferation,migration and invasion of SKOV3cells(P<0.05).High levels of exosomal miR-145 decreased the proliferation,migration and proliferation of SKOV3 cells,inhibited angiogenesis in human umbilical vein endothelial cells,and suppressed tumor sphere forming capacity of SKVO3cells(P<0.05).However,miR-145-downregula-ted exosomes had opposite effects(P<0.05).Conclusion:The transfer of miR-145 through exosomes can inhibit the migration,invasion and angiogenesis of epithelial ovarian cancer cells,and regulate the stem cell-like characteristics of epithelial ovarian cancer.
作者
张文晶
王静
张玉瑾
余明莲
王辉
Wenjing Zhang;Jing Wang;Yujin Zhang;Minglian Yu;Hui Wang(Department of Obstetrics and Gynecology,The Seventh Medical Center of PLA General Hospital,Beijing 100700,China)
出处
《广西医科大学学报》
CAS
2021年第11期2019-2027,共9页
Journal of Guangxi Medical University
基金
funded by National Natural Science Foundation of China(No.81341075)。
