CAFs来源外泌体miR-3173-5p靶向ACSL4铁死亡途径调节口腔鳞癌细胞恶性表型  

The regulatory role of exosome-derived miR-3173-5p in targeting the ACSL4 ferroptosis pathway to modulate the malignant phenotype of oral squamous cell carcinoma cells

作　　者：邴利 周乐乐 陈玉[1] 金和 BING Li;ZHOU Le-le;CHEN Yu;JIN He(Department of Stomatology,Benq Hospital Affiliated to Nanjing Medical University,Jiangsu Nanjing 210019,China)

机构地区：[1]南京医科大学附属明基医院口腔科,江苏南京210019

出　　处：《临床口腔医学杂志》2024年第6期331-338,共8页Journal of Clinical Stomatology

基　　金：江苏省卫生健康委员会科研项目(202011364)。

摘　　要：目的:探讨肿瘤相关成纤维细胞(cancer-associated fibroblasts, CAFs)来源外泌体miR-3173-5p对口腔癌的恶性表型的影响及作用机制。方法:原代培养、分离、纯化、验证正常成纤维细胞(normal fibroblasts, NFs)和CAFs,收集各组细胞上清外泌体,透射电镜和纳米颗粒示踪分析观察和鉴定外泌体。RT-qPCR检测miR-3173-5p及铁死亡指标ACSL4的mRNA的表达。CCK-8、EdU实验、克隆形成实验、划痕实验和Transwell实验分别检测细胞活性、增殖、迁移和侵袭情况。双荧光素酶报告基因系统用于检测miR-3173-5p与ACSL4的相关性。Western blot检测外泌体相关标志物(HSP70、CD81、CD9、CD63、TSG101和Alix)及ACSL4蛋白表达。异种移植瘤实验验证CAFs来源外泌体miR-3173-5p在口腔癌中的促进作用。结果:口腔癌患者组织来源的CAFs外泌体中miR-3173-5p表达明显升高,促进口腔癌细胞增殖、迁移和侵袭。miR-3173-5p可直接靶向ACSL4。过表达ACSL4减弱miR-3173-5p对口腔癌细胞恶性表型的促进作用。体内实验证实外泌体miR-3173-5p促进口腔癌肿瘤的生长。结论:CAFs来源外泌体miR-3173-5p通过靶向铁死亡指标ACSL4驱动口腔癌细胞的恶性表型。Objective:An investigation was conducted to determine weather exosome miR-3173-5p from cancer-associated fibroblasts(CAFs) affects the malignant phenotype of oral cancer and its mechanisms.Methods:The initial step in this study involved establishing primary cultures of normal fibroblasts(NFs)and CAFs, followed by isolation, purification, and verification.Subsequently, exosomes were collected and identified using Transmission Electron Microscope(TEM)and nanoparticle tracers.The expression of miR-3173-5p and ACSL4 mRNA,a marker associated with ferroptosis, was analyzed using RT-qPCR.Cell activity, proliferation, migration, and invasion were assessed through various assays including CCK-8,EdU incorporation, clonogenesis, scratch, and Transwell assays.MiR-3173-5p and ACSL4 were detected using the dual luciferase reporter gene system.Detection of exosomal marker proteins(HSP70,CD81,CD9,CD63,TSG101,and Alix)and expression of ACSL4.Experiments on xenograft tumors confirm the promoting role of miR-3173-5p on oral cancer exosomes derived from CAFs.Results:CAFs exosomes derived from patients with oral cancer were significantly overexpressed with miR-3173-5p, resulting in cell proliferation, migration, and invasion.Oral cancer cells overexpressing ACSL4 have reduced miR-3173-5p's capacity to promote malignant phenotypes.An in vivo study has shown that the exosome miR-3173-5p promotes tumor growth in oral cancers.Conclusion:By targeting ACSL4,miR-3173-5p from CAFs-derived exosomes promotes the malignant phenotype of oral cancer cells.

关 键 词：口腔癌 肿瘤微环境 外泌体 miR-3173-5p 铁死亡 恶性表型 

分 类 号：R739.8[医药卫生—肿瘤]