摘要
目的探讨骨肉瘤外泌体对肿瘤相关成纤维细胞转化的影响及机制。方法提取并鉴定骨肉瘤细胞U2-OS、MG-63,人成骨细胞hFoB1.19外泌体,将PBS、hFoB1.19、U2-OS、MG-63外泌体与人肺成纤维细胞MRC-5共孵育,CCK8法检测MRC-5增殖能力,Transwell小室法检测MRC-5迁移和侵袭能力,qRT-PCR检测MRC-5细胞中炎症因子IL-1β、IL-6、IL-8表达,Western blot检测MRC-5细胞α-SMA、Vimentin、FAP表达,JAK2/STAT3信号通路的激活。结果透射电镜和Western blot鉴定所提取的外泌体符合其结构特征;与U2-OS、MG-63外泌体共孵育后,MRC-5增殖、迁移和侵袭能力显著增加,MRC-5中IL-1β、IL-6、IL-8表达显著增加,α-SMA、Vimentin、FAP表达显著增加,JAK2和STAT3磷酸化水平显著增加(均P<0.001)。结论骨肉瘤细胞外泌体通过激活JAK2/STAT3信号通路而促进成纤维细胞向肿瘤相关成纤维细胞转化。
Objective To investigate the effect and mechanism of osteosarcoma exosomes on tumor-associated fibroblast transformation.Methods The exosomes of osteosarcoma cells U2-OS,Mg-63 and human osteoblasts hfob1.19 were extracted and identified.The exosomes of PBS,hFoB1.19,U2-OS and Mg-63 were incubated with human lung fibroblasts MRC-5.The proliferation ability of MRC-5 was detected by CCK8 method,the migration and invasion ability of MRC-5 was detected by Transwell chamber method,and the inflammatory factor IL-1 in MRC-5 cells was detected by QRT-PCR-β、The expression of IL-6 and IL-8 was detected by Western blotαThe expression of SMA,vimentin and FAP,and the activation of JAK2/STAT3 signaling pathway.Results he exosomes were identified by transmission electron microscope and Western blot.When CO incubated with U2-OS and Mg-63 exosomes,the proliferation,migration and invasion of MRC 5 increased significantly;IL-1β,IL-6 and IL-8 in MRC 5 increased significantly;the expressions ofα-SMA,vimentin and FAP were significantly increased,and the phosphorylation levels of JAK2 and STAT3 were significantly increased(all P<0.001).Conclusion Osteosarcoma cell exosomes promote the transformation of fibroblasts into tumor-associated fibroblasts by activating JAK2/STAT3 signaling pathway.
作者
折胜利
宋兴华
周杨
何园
贾晶
刘彦博
王良华
ZHE Shengli;SONG Xinghua;ZHOU Yang;HE Yuan;JIA Jing;LIU Yanbo;WANG Lianghua(Department of Orthopedics, Northwestern Polytechnical University Hospital, Xi'an 710012, China;Department of Osteopathy/Bone Oncology, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, China;Department of Orthopedics, Xin'an Central Hospital of Shaanxi Province, Xi'an 710048, China;School of Basic Medicine, Xinjiang Medical University,Urumqi 830011, China)
出处
《西部医学》
2021年第8期1096-1100,1105,共6页
Medical Journal of West China
基金
国家自然科学基金地区科学基金项目(81860394/H0607)。
