摘要
目的 探讨骨肉瘤细胞来源的外泌体诱导肿瘤相关成纤维细胞生成的相关机制。方法 将提取并鉴定好的骨肉瘤细胞外泌体与人肺成纤维细胞共培养,通过RT-qPCR和Western blot方法检测肿瘤相关成纤维细胞特异性蛋白α-SMA、FAP的表达;Transwell方法检测共培养后成纤维细胞迁移侵袭能力。生物信息学分析骨肉瘤患者与健康人群血浆中差异表达的LncRNA,找到差异表达显著的LncRNA NCK1-DT,再通过qRT-PCR方法检测LncRNA NCK1-DT在骨肉瘤细胞外泌体和成骨细胞外泌体的表达情况。敲低骨肉瘤细胞外泌体中LncRNA NCK1-DT的表达并再次与成纤维细胞共培养,检测共培养后α-SMA、FAP表达的变化,以及成纤维细胞迁移侵袭能力的变化。结果 骨肉瘤细胞外泌体在与成纤维细胞共培养后,成纤维细胞内α-SMA、FAP的表达明显增加,且成纤维细胞的迁移和侵袭能力明显增强。敲低骨肉瘤细胞外泌体中LncRNA NCK1-DT的表达后,成纤维细胞中α-SMA、FAP的表达显著下降,同时成纤维细胞的迁移和侵袭能力也显著降低。结论 骨肉瘤细胞可通过分泌并传递携带LncRNA NCK1-DT的外泌体,诱导成纤维细胞的转化,促进肿瘤相关成纤维细胞的生成。
Objective To investigate the mechanism by which osteosarcoma-derived exosomes induce the gener-ation of cancer-associated fibroblasts(CAFs).Methods Human lung fibroblasts were co-cultured with isolated and characterized osteosarcoma-derived exosomes.The expressions of CAFs-specific markersα-smooth muscle actin(α-SMA)and fibroblast activation protein(FAP)were analyzed by using RT-qPCR and Western blot.Transwell assays were performed to evaluate the migratory and invasive capacities of co-cultured fibroblasts.Bioinformatic analysis identi-fied differentially expressed LncRNAs between osteosarcoma patients and healthy controls,with LncRNA NCK1-DT showing significant upregulation.qRT-PCR was employed to compare LncRNA NCK1-DT levels in osteosarcoma-de-rived exosomes versus osteoblast-derived exosomes.Knockdown of LncRNA NCK1-DT in osteosarcoma exosomes was followed by co-culture with fibroblasts to assess subsequent changes inα-SMA/FAP expression and fibroblast migra-tion/invasion.Results Co-culture with osteosarcoma exosomes significantly upregulatedα-SMA and FAP expression in fibroblasts while enhancing their migratory and invasive capacities.LncRNA NCK1-DT knockdown in osteosarcoma exosomes markedly reducedα-SMA/FAP expression and attenuated fibroblasts migration/invasion.Conclusion Osteo-sarcoma cells promote fibroblast transformation and CAFs generation through exosomal transfer of LncRNA NCK1-DT.
作者
王艺为
张辉
姚佩峰
滕松龄
WANG Yi-wei;ZHANG Hui;YAO Pei-feng;TENG Song-ling(Department of Hand Surgery,Central Hospital affiliated to Shenyang Medical College,Shenyang 110024,China)
出处
《解剖科学进展》
2025年第6期811-815,共5页
Progress of Anatomical Sciences
基金
辽宁省科学技术计划项目面上项目(2022-MS-411)。
