M2-like tumor-associated macrophages-secreted Wnt1 and Wnt3a promotes dedifferentiation and metastasis via activating β-catenin pathway in thyroid cancer
M2-like tumor-associated macrophages-secreted Wnt1 and Wnt3a promotes dedifferentiation and metastasis via activating β-catenin pathway in thyroid cancer
复制标题
M2类肿瘤相关巨噬细胞分泌的Wnt1和Wnt3a通过激活甲状腺癌中的β-连环蛋白途径促进去分化和转移
DOI：
10.1002/mc.23268
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发表时间：
2020-12-07
影响因子：
4.6
通讯作者：
Deng, Zhi-Yong
中科院分区：
文献类型：
--
作者：
Lv, Juan;Feng, Zhi-Ping;Deng, Zhi-Yong
背景甲状腺癌(TC)的发病率在全球范围内迅速上升，已成为一个全球性的问题。尽管大多数TC侵袭性不强，预后良好，但对间变性TC的治疗相对有限，其机制尚不清楚。用流式细胞术鉴定细胞培养中M2样肿瘤相关巨噬细胞(TAMs)的表面标志。采用实时定量聚合酶链式反应、免疫印迹分析、免疫组织化学等方法检测Wnt1、Wnt3a、Wnt/β-catenin途径各组分及增殖/上皮-间充质转化(EMT)相关蛋白的表达。应用碱性磷酸酶活性测定、克隆形成实验和Transwell实验分别检测Wnt1、Wnt3a和β-catenin途径在TC细胞去分化、增殖、迁移和侵袭中的作用。结果与M2样TAMs共培养可促进TC细胞的去分化、增殖、迁移和侵袭。在共培养的TC细胞中，EMT和增殖相关蛋白也被促进。在共培养体系中，WNT1和WNT3a的水平升高。阻断WNT1或WNT3a可抑制共培养肿瘤细胞的恶性行为。此外，Wnt1或Wnt3a基因敲除抑制了Wnt/β-catenin信号通路，并抑制了共培养肿瘤细胞的EMT和增殖相关信号。结论M2样TAMs通过分泌Wnt1和Wnt3a，进而激活β-连环素，促进TC的脱分化、增殖和转移。
Background Thyroid carcinoma (TC) has been a global issue for its rapid increasing incidence worldwide. Although most TC was not so aggressive with a good prognosis, treatment against anaplastic TC was relatively limited and the mechanisms are not well elucidated yet.Methods TC cell lines (IHH4 and TPC-1) were used. Flow cytometry was used to identify the surface marker of M2-like tumor-associated macrophages (TAMs) from cell culture. Quantitative real-time polymerase chain reaction, western blot analysis, immunostaining, and immunohistochemistry were used to detect the expression of Wnt1, Wnt3a, components of Wnt/beta-catenin pathway, and proliferation/epithelial-mesenchymal transition (EMT)-related proteins. Alkaline phosphatase activity assay, colony formation assay, and transwell assay were used to examine the roles of Wnt1, Wnt3a, and beta-catenin pathway in cell dedifferentiation, proliferation, migration, and invasion of TC cells, respectively. Subcutaneous tumor growth was monitored in nude mice.Results Coculture with M2-like TAMs facilitated dedifferentiation, proliferation, migration, and invasion in TC cells. EMT and proliferation-related proteins were also promoted in cocultured TC cells. The level of Wnt1 and Wnt3a was increased in the coculture system. Block of Wnt1 or Wnt3a suppressed malignant behaviors in cocultured tumor cells. Furthermore, Wnt1 or Wnt3a knockdown inhibited Wnt/beta-catenin signaling pathway, and suppressed EMT and proliferation-related signals in cocultured tumor cells. Knockdown of Wnt1 or Wnt3a inhibited tumor growth in xenograft model.Conclusion M2-like TAMs promoted dedifferentiation, proliferation, and metastasis of TC by Wnt1 and Wnt3a secretion and ensuing beta-catenin activation.