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首页|期刊导航|中国医学前沿杂志(电子版)|经血间充质干细胞外泌体对人卵巢癌细胞A2780生物学行为的影响

经血间充质干细胞外泌体对人卵巢癌细胞A2780生物学行为的影响OACSTPCD

Effects of exosomes derived from menstrual blood-derived mesenchymal stem cells on biological behavior of human ovarian cancer cell line A2780

中文摘要英文摘要

目的 探讨经血源性间充质干细胞(menstrual blood-derived mesenchymal stem cells,MenSCs)来源的外泌体(MenSCs-derived exosomes,MenSCs-Exos)对人卵巢癌细胞 A2780 增殖、迁移、侵袭能力和细胞周期分布的影响和机制.方法 分离纯化MenSCs,流式细胞术鉴定其细胞表型,油红O 染色观察其成脂分化能力;超速离心法分离MenSCs-Exos,电子显微镜观察外泌体形态,纳米颗粒追踪分析其大小及分布,蛋白免疫印迹法检测外泌体标志性蛋白分子 CD9、TSG101、calnexin的表达.采用CCK-8 实验、划痕实验、Transwell细胞体外侵袭实验、流式细胞仪检测 MenSCs-Exos 对A2780 细胞增殖、迁移、侵袭能力和细胞周期分布的影响;蛋白免疫印迹法检测Wnt/β-catenin信号通路相关蛋白和迁移相关蛋白的表达.结果 成功分离具有干细胞表型和成脂分化潜能的 MenSCs,同时检测发现 MenSCs-Exos 为边缘清晰、大小分布均匀的膜性囊泡,粒径为 30~150 nm,平均为 72.89 nm,膜表面标志性蛋白 CD9、TSG101 表达呈阳性,calnexin表达呈阴性.与对照组相比,MenSCs-Exos能增强 A2780 细胞增殖、迁移和侵袭能力,影响其细胞周期分布,并且可上调β-catenin、c-Myc、TCF4、CyclinD1、Vimentin、N-cadherin的表达,下调 E-cadherin的表达.结论 MenSCs-Exos 可能是通过激活 Wnt/β-catenin信号通路促进 A2780 细胞增殖,并通过下调 E-cadherin的表达促进 A2780 细胞的迁移和侵袭.

Objective To investigate the effect and mechanism of menstrual blood-derived mesenchymal stem cells(MenSCs)derived exosomes(MenSCs-Exos)on proliferation,migration,invasion and cell cycle distribution of human ovarian cancer cell line A2780.Methods MenSCs were isolated and purified,cell phenotype was identified by flow cytometry,adipogenic differentiation ability was observed by red oil O staining,MenSCs-Exos were isolated by ultracentrifugation,the morphology of exosomes were observed by electron microscope,the size and distribution were analyzed by NTA,and the expression of CD9,calnexin and TSG101 were detected by Western blot.The effects of MenSCs-Exos on the proliferation,migration,invasion and cell cycle distribution of A2780 cells were detected by CCK-8 assay,wound healing assay,Transwell migration and invasion assay and flow cytometry.The expression of Wnt/β-catenin signaling pathway related proteins and migration related proteins were detected by Western blot.Results MenSCs with stem cell phenotype,osteogenic and adipogenic differentiation potential were successfully isolated.At the same time,MenSCs-Exos was found a membranous vesicle with clear edge and uniform size distribution,the diameter of which was about 30-150 nm,with an average of 72.89 nm.The expression of CD9 and TSG101 on the membrane surface was positive,while the expression of calnexin was negative.Compared with the control group,MenSCs-Exos treatment group could enhance the ability of proliferation,migration and invasion of A2780 cells,affect the distribution of cell cycle and up-regulate the expression of β-catenin,c-Myc,TCF4,CyclinD1,Vimentin and N-cadherin,and down-regulate the expression of E-cadherin.Conclusion MenSCs-Exos may promote the proliferation of ovarian cancer cells by activating Wnt/β-catenin signal pathway and promote the migration and invasion of A2780 cells by up-regulating the expression of E-cadherin.

陆茜林;丁娜;贺红林;袁怡君;贺天文;冯露;田超;刘康;李均

川北医学院第二临床医学院 (南充市中心医院)妇科,四川 南充 637000川北医学院第二临床医学院 (南充市中心医院)烧伤整形美容外科,四川 南充 637000川北医学院第二临床医学院 (南充市中心医院)组织工程与干细胞研究所,四川 南充 637000

外泌体经血源间充质干细胞卵巢癌Wnt/β-catenin信号通路

ExosomesMenstrual blood-derived mesenchymal stem cellsOvarian cancerWnt/β-catenin signal pathway

《中国医学前沿杂志(电子版)》 2024 (002)

50-57 / 8

南充市校科技战略合作项目(22SXQT0134) Nanchong Municipal School of Science and Technology Strategic Cooperation Special Project(22SXQT0134)

10.12037/YXQY.2024.02-08

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