姜黄素介导的骨髓间充质干细胞外泌体对ACC-M细胞增殖、迁移和侵袭的影响  

Curcumin-mediated exosomes from bone marrow mesenchymal stem cells inhibits ACC-M cell proliferation,migration and invasion in vitro

作　　者：程昊 吴发印 刘智丹 CHENG Hao;WU Fayin;LIU Zhidan(Department of Oral and Maxillofacial Surgery,Fifth Affiliated(Zhuhai)Hospital,Zunyi Medical University,Zhuhai 519100,China)

机构地区：[1]遵义医科大学第五附属(珠海)医院口腔颌面外科,珠海519100

出　　处：《口腔颌面外科杂志》2024年第1期34-39,共6页Journal of Oral and Maxillofacial Surgery

摘　　要：目的:探讨姜黄素介导的骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)外泌体对ACC-M细胞增殖和转移的影响及其作用机制。方法:将7.5μmmol/L的姜黄素与人BMSCs共培养72 h后,分离外泌体并进行鉴定,将外泌体与ACC-M细胞进行共培养,并将ACC-M细胞分为control组(正常培养)、BMSC-Exo组(未经姜黄素处理的BMSCs外泌体与ACC-M细胞共培养)、Cur-BMSC-Exo组(经姜黄素处理的BMSCs外泌体与ACC-M细胞共培养)及Cur组(经7.5μmmol/L的姜黄素处理细胞);培养48 h后使用蛋白质印迹法(Western blotting)实验检测外泌体肿瘤易感基因101(tumor susceptibility gene 101,TSG-101)、CD63、CD9和重组人钙连蛋白(calnexin,CANX)及ACC-M细胞上皮钙黏素(E-cadherin)、神经钙黏素(N-cadherin)、波形蛋白(vimentin)和转化生长因子β1(transforming growth factor-β1,TGF-β1)、细胞外信号调节激酶(extracellular signal-regulated kinase,ERK)的蛋白表达情况;使用细胞计数试剂盒-8(cell counting kit-8,CCK8)实验检测ACC-M细胞存活率;使用transwell实验检测ACC-M细胞迁移和侵袭情况;使用实时定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测ACC-M细胞TGF-β1和ERK mRNA相对表达量。结果:与control组和BMSC-Exo组比较,Cur-BMSC-Exo组和Cur组的细胞活力、细胞迁移和侵袭数量及N-cadherin、vimentin、TGF-β1和ERK表达水平均显著降低(P<0.05),而E-cadherin蛋白水平显著增加(P<0.05);与Cur-BMSC-Exo组比较,Cur组细胞活力、细胞迁移和侵袭数量及N-cadherin、vimentin、TGF-β1和ERK表达水平均显著增加(P<0.05),而E-cadherin蛋白水平显著降低(P<0.05)。结论:BMSCs分泌的外泌体可作为姜黄素的载体,抑制ACC-M细胞的增殖和转移并调控TGF-β1/ERK信号通路。Objective:To investigate the effect of curcumin-mediated exosomes of bone marrow mesenchymal stem cells(BMSCs)on proliferation and metastasis of ACC-M cells and its mechanism.Methods:After being co-cultured with 7.5μmmol/L curcumin and BMSCs for 72 h,exosomes were isolated and identified,and co-cultured with ACC-M cells.ACC-M cells were divided into control group(normal culture),BMSC-Exo group(no curcumin-treated BMSCs exosomes co-cultured with ACC-M cells),Cur-BMSC-Exo group(curcumin-treated BMSCs exosomes co-cultured with ACC-M cells)and Cur group(7.5μmmol/L curcumin treated cells).Western blotting was used to detect the expression of tumor susceptibility gene 101(TSG-101),CD63,CD9 and recombinant human calnexin(CANX)proteins in exosome and the expression of E-cadherin,N-cadherin,vimentin,transforming growth factor-β1(TGF-β1)and p-ERK proteins in ACC-M cells after 48 h of culture.The survival rate of ACC-M cells was determined by cell counting kit-8(CCK-8)assay.The migration and invasion of ACC-M cells were detected by transwell assay.The relative expression levels of TGF-β1 and ERK mRNA in ACC-M cells were detected by real-time quantitative polymerase chain reaction(RT-qPCR)assay.Results:Compared with control group and BMSC-Exo group,the cell viability,cell migration and invasion number,and the expression levels of N-cadherin,vimentin,TGF-β1 and ERK were significantly decreased in Cur-BMSC-Exo and Cur groups(P<0.05),while E-cadherin protein level was significantly increased(P<0.05);compared with the Cur-BMSC-Exo group,the cell viability,cell migration and invasion numbers,and the expression levels of N-cadherin,vimentin,TGF-β1 and ERK were significantly increased in the Cur group(P<0.05),while E-cadherin protein level was significantly decreased(P<0.05).Conclusion:BMSCs exosomes can act as curcumin carrier to inhibit proliferation and metastasis of ACC-M cells and regulate TGF-β1/ERK signaling pathway.

关 键 词：姜黄素 骨髓间充质干细胞 外泌体 ACC-M 增殖 

分 类 号：R782[医药卫生—口腔医学]