《Journal of Oral and Maxillofacial Surgery》 ›› 2024, Vol. 34 ›› Issue (1): 34-39. doi: 10.12439/kqhm.1005-4979.2024.01.005

• Basic Scientific Study • Previous Articles     Next Articles

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

CHENG Hao(), WU Fayin(), LIU Zhidan   

  1. Department of Oral and Maxillofacial Surgery, Fifth Affiliated (Zhuhai) Hospital, Zunyi Medical University, Zhuhai 519100, China
  • Received:2022-08-08 Accepted:2023-03-31 Online:2024-02-28 Published:2024-02-28

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

程昊(), 吴发印(), 刘智丹   

  1. 遵义医科大学第五附属(珠海)医院口腔颌面外科,珠海 519100
  • 通讯作者: 吴发印,主任医师. E-mail:dududu202112@163.com
  • 作者简介:
    程昊,主治医师. E-mail:

Abstract:

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 7.5 μmmol/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 (7.5 μmmol/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.

Key words: curcumin, bone marrow mesenchymal stem cells, exosome, ACC-M, proliferation

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7.5 µmmol/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7.5 µmmol/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。

关键词: 姜黄素, 骨髓间充质干细胞, 外泌体, ACC-M, 增殖

CLC Number: