Exosomal Dvl3 promoted the aggressive phenotypic transformation of RA-FLS via wnt pathway
Exosomal Dvl3 promoted the aggressive phenotypic transformation of RA-FLS via wnt pathway
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DOI：
10.1080/08916934.2022.2067984
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发表时间：
2022-05
期刊：
影响因子：
3.5
通讯作者：
Weixing Tan;N. Chen;Y. Qiu;Xiaomei Feng;Shuwen Li;Yongjin Zhang;Haoran Li;Jie Gao;Dongbao Zhao
中科院分区：
文献类型：
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作者：
Weixing Tan;N. Chen;Y. Qiu;Xiaomei Feng;Shuwen Li;Yongjin Zhang;Haoran Li;Jie Gao;Dongbao Zhao
【摘要】目的通过外泌体干预，探讨Dvl3在RA-FLS中的作用及机制。方法采用免疫组化法、WB法、qPCR法检测Dvl3的表达谱。将转染了Dvl3过表达慢病毒（OE）的RA-FLS培养上清获得的修饰外泌体注入目标RA-FLS。分别采用CKK8试剂盒、Tunel、迁移试验、qPCR和酶联免疫吸附试验（ELISA）检测外泌体Dvl3影响的存活能力、迁移能力和炎症因子的产生；流式细胞术分析外泌体对CD4+ T细胞的炎症调节作用。通过qPCR和WB筛选Dvl3可能的下游通路，并通过双荧光素酶报告基因实验进行验证。结果RA和CIA组织中Dvl3表达水平明显升高。OE组外泌体能显著促进细胞增殖活性、迁移/侵袭能力。外泌体Dvl3-OE组TNF-α、IL-1β、IL-17、IL-21水平升高。外泌体Dvl3增强了Th1和Th17细胞的极化和相关的细胞因子。过表达Dvl3的同时，β-catenin和RhoA活性显著升高。结论本研究发现RA患者外泌体中Dvl3的高表达可能具有通过Wnt途径促进RA- fls表型转化的能力。
Abstract Objective This study was performed to explore the function and mechanism of Dvl3 in RA-FLS by exosome intervention. Methods The expression pattern of Dvl3 was examined by IHC, WB, and qPCR. Modified exosomes obtained from culturing supernatant of RA-FLS infected with Dvl3 over expression (OE) lentivirus were administrated to the target RA-FLS. The ability of survival, migration, and the production of inflammatory factor influenced by exosomal Dvl3 were detected by CKK8 kits, Tunel, migration test, qPCR, and enzyme-linked immunosorbent assay (ELISA) respectively; Flow cytometry analysis was conducted to explorer the inflammatory moderate role of exosomes on CD4+ T cells. The possible downstream pathways of Dvl3 were screened by qPCR and WB and verified by double luciferase reporter experiment. Results The expression level of Dvl3 was significantly increased in RA and CIA. Exosomes from the OE group could significantly promote cell proliferation activity, migration/invasion ability. The augment of TNF-α, IL-1β, IL-17, and IL-21 was observed in exosomal Dvl3-OE group. Th1 and Th17 cells polarisation and cytokines related were both enhanced by Exosomal Dvl3. Over expression of Dvl3 was accompanied by the significant increase of β-catenin and RhoA activities. Conclusion This study discovered the high expression of Dvl3 of exosomes derived from RA patients which may possessed the ability to promote phenotypic transformation of RA-FLS through Wnt pathway.