M2-like tumour-associated macrophage-secreted IGF promotes thyroid cancer stemness and metastasis by activating the PI3K/AKT/mTOR pathway
M2-like tumour-associated macrophage-secreted IGF promotes thyroid cancer stemness and metastasis by activating the PI3K/AKT/mTOR pathway
复制标题
M2样肿瘤相关巨噬细胞分泌的IGF通过激活PI3K/AKT/mTOR通路促进甲状腺癌干细胞和转移
DOI：
10.3892/mmr.2021.12249
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发表时间：
2021-08-01
影响因子：
3.4
通讯作者：
Deng, Zhi-Yong
中科院分区：
文献类型：
--
作者：
Lv, Juan;Liu, Chao;Deng, Zhi-Yong
已证实M2样肿瘤相关巨噬细胞(TAMs)促进间变性甲状腺癌(ATC)的生长。然而，ATC中M2样TAMs的潜在机制仍不清楚。因此，本研究旨在探讨M2样TAMs在ATC中的作用及其机制。用PMA+IL-4、IL-13诱导M2样TAMs，并用流式细胞仪鉴定。Transwell法和球形法检测ATC细胞的侵袭性和干性。用酶联免疫吸附试验和逆转录-定量聚合酶链式反应检测胰岛素样生长因子-1和胰岛素样生长因子-2的表达水平。Western blotting检测与上皮-间充质转化(EMT)、干性和PI3K/AKT/mTOR通路相关的蛋白。采用免疫组织化学方法检测M2样标志物CD68和CD206在甲状腺癌组织和甲状腺腺瘤组织中的表达。结果表明，PMA联合IL-4和IL-13可成功地诱导出类M2 TAMs。与类M2细胞共培养后，ATC细胞的侵袭能力和干细胞性显著增强。EMT相关标志物N-cadherin和Vimentin，干性相关标志物Oct4、Sox2和CD133以及胰岛素受体(IR)-A/IGF1受体(IGF1R)的表达水平显著上调，而E-cadherin的表达水平显著降低。此外，胰岛素样生长因子-1和胰岛素样生长因子-2的产生显著增加。值得注意的是，外源性IGF-1/IGF-2促进了C643细胞的侵袭和干化，而阻断IGF-1和IGF-2则通过抑制IR-A/IGF-1R介导的PI3K/AKT/mTOR信号通路来抑制共培养体系中的转移和干化。免疫组化结果显示CD68和CD206在ATC组织中的表达明显增强。综上所述，M2样TAMs促进了ATC细胞的转移，增加了ATC细胞的干性，其机制可能与M2样TAMs切割IGF，激活IR-A/IGF1R介导的PI3K/AKT/mTOR信号通路有关。
M2-like tumour-associated macrophages (TAMs) have been demonstrated to promote the growth of anaplastic thyroid carcinoma (ATC). However, the underlying mechanism of M2-like TAMs in ATC remains unclear. Thus, in the present study, the role and mechanism of M2-like TAMs in ATC were investigated. M2-like TAMs were induced by treatment with PMA, plus IL-4 and IL-13, and identified by flow cytometry. Transwell and sphere formation assays were applied to assess the invasion and stemness of ATC cells. The expression levels of insulin-like growth factor (IGF)-1 and IGF-2 were examined by ELISA and reverse transcription-quantitative PCR. Proteins related to the epithelial-mesenchymal transition (EMT), stemness and the PI3K/AKT/mTOR pathway were examined via western blotting. Immunohistochemistry (IHC) was used to detect the expression of the M2-like TAM markers CD68 and CD206 in ATC tissues and thyroid adenoma tissues. It was found that treatment with PMA plus IL-4 and IL-13 successfully induced M2-like TAMs. Following co-culture with M2-like TAMs, the invasive ability and stemness of ATC cells were significantly increased. The expression levels of the EMT-related markers N-cadherin and Vimentin, the stemness-related markers Oct4, Sox2 and CD133, and the insulin receptor (IR)-A/IGF1 receptor (IGF1R) were markedly upregulated, whereas E-cadherin expression was significantly decreased. In addition, the production of IGF-1 and IGF-2 was significantly increased. Of note, exogenous IGF-1/IGF-2 promoted the invasion and stemness of C643 cells, whereas blocking IGF-1 and IGF-2 inhibited metastasis and stemness by repressing IR-A/IGF-1R-mediated PI3K/AKT/mTOR signalling in the co-culture system. IHC results showed that the expression of CD68 and CD206 was obviously increased in ATC tissues. To conclude, M2-like TAMs accelerated the metastasis and increased the stemness of ATC cells, and the underlying mechanism may be related to the section of IGF by M2-like TAMs, which activates the IR-A/IGF1R-mediated PI3K/AKT/mTOR signalling pathway.