口腔癌细胞来源外泌体miR-582-3p通过靶向SFRP1驱动口腔癌细胞的恶性表型  

Oral cancer cell-derived exosomal microRNA-582-3p promotes the malignant behaviors of oral cancer cells by targeting secreted frizzled-related protein 1

作　　者：彭德瑞 钟晓敏 PENG De-rui;ZHONG Xiao-min(Oral and Maxillofacial Surgery Department of Changsha Stomatological Hospital,Hunan Changsha 410004,China;Department of Stomatology,First Affiliated Hospital of Gannan Medical College,Jiangxi Ganzhou 341000,China)

机构地区：[1]长沙市口腔医院口腔颌面外科,湖南长沙410004 [2]赣南医学院第一附属医院口腔科,江西赣州341000

出　　处：《临床口腔医学杂志》2024年第2期67-74,共8页Journal of Clinical Stomatology

基　　金：湖南省创新型省份建设专项(S2019JJKWLH1721)。

摘　　要：目的:探讨口腔癌细胞来源外泌体miR-582-3p对口腔癌细胞的恶性表型的影响及作用机制。方法:收集口腔患者血清和细胞上清外泌体,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microRNA-582-3p(miR-582-3p)及分泌卷曲相关蛋白1(secreted frizzled-related protein 1,SFRP1)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-582-3p与SFRP1的相关性。Western blot检测外泌体相关标志物(HSP70、CD81、CD9、CD63、TSG101和Alix)及SFRP1蛋白表达。异种移植瘤实验验证口腔癌细胞来源外泌体miR-582-3p在口腔癌中的促进作用。结果:口腔癌患者组织、血清及血清和细胞外泌体中miR-193b-3p表达明显升高。口腔癌细胞外泌体miR-582-3p促进口腔癌细胞增殖、迁移和侵袭。miR-582-3p可直接靶向SFRP1。过表达SFRP1减弱miR-582-3p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-582-3p促进口腔癌肿瘤的生长。结论:口腔癌细胞来源外泌体miR-582-3p通过靶向SFRP1驱动口腔癌细胞的恶性表型。Objective:To explore the supportive effects of exosomal microRNA-582-3p(miR-582-3p)on the malignant behaviors of oral cancer cells and its potential mechanism.Methods:Oral cancer serum and cell media were collected for exosome isolation.Transmission electron microscopy and nanoparticle tracking analysis were performed for exosome identification and concentration and diameters measurement.miR-582-3p level and secreted frizzled-related protein 1(SFRP1)mRNA expression were detected by quantitative real-time PCR.Oral cancer cell viability,proliferation,migration,and invasion were assessed by cell count kit-8(CCK-8),EdU,colony formation,wound healing,and Transwell assays.The direct interaction between miR-582-3p and SFRP1 was determined by dual-luciferase activity assay.Western blot analysis was performed to detect exosome-related proteins,including HSP70,CD81,CD9,CD63,TSG101 and Alix,as well as SFRP1 protein expression levels.A xenograft experiment was utilized to confirm the tumor-promoting role of exosomal miR-582-3p in oral cancer.Results:miR-582-3p was highly expressed in oral cancer tissues,serum,cells and exosomes derived from serum and cells.Oral cancer cell-derived exosomal miR-582-3p significantly promoted the viability,proliferation,migration,and invasion of oral cancer cells.Dual-luciferase reporter assay revealed that miR-582-3p can directly bind to the 3′-UTR of SFRP1 and inhibit its expression.Restoration of SFRP1 expression rescued the malignant phenotypes induced by exosomal miR-582-3p.In addition,enforced expression of miR-582-3p facilitated oral cancer growth in a xenograft model,accompanied by downregulation of SFRP1.Conclusion:Oral cancer cell-derived exosomal miR-582-3p promotes the malignant behaviors of oral cancer cells by targeting SFRP1.

关 键 词：口腔癌 外泌体 miR-582-3p 分泌卷曲相关蛋白1(SFRP1) 

分 类 号：R739.8[医药卫生—肿瘤]