Department of Nuclear Medicine, The Third Affiliated Hospital of Kunming Medical University, Kunming, Yunnan 650118, P.R. China.
Mol Med Rep. 2021 Aug;24(2). doi: 10.3892/mmr.2021.12249. Epub 2021 Jun 29.
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.
M2 样肿瘤相关巨噬细胞(TAMs)已被证实可促进间变性甲状腺癌(ATC)的生长。然而,M2 样 TAMs 在 ATC 中的潜在机制尚不清楚。因此,在本研究中,研究了 M2 样 TAMs 在 ATC 中的作用和机制。通过用 PMA 加 IL-4 和 IL-13 处理来诱导 M2 样 TAMs,并通过流式细胞术进行鉴定。Transwell 和球体形成实验用于评估 ATC 细胞的侵袭和干性。通过 ELISA 和逆转录定量 PCR 检测胰岛素样生长因子(IGF)-1 和 IGF-2 的表达水平。通过 Western blot 检测上皮-间充质转化(EMT)、干性和 PI3K/AKT/mTOR 通路相关蛋白的表达。免疫组织化学(IHC)用于检测 ATC 组织和甲状腺腺瘤组织中 M2 样 TAM 标志物 CD68 和 CD206 的表达。结果发现,用 PMA 加 IL-4 和 IL-13 处理成功诱导 M2 样 TAMs。与 M2 样 TAMs 共培养后,ATC 细胞的侵袭能力和干性明显增加。EMT 相关标志物 N-钙粘蛋白和波形蛋白、干性相关标志物 Oct4、Sox2 和 CD133 以及胰岛素受体(IR)-A/IGF1 受体(IGF1R)的表达明显上调,而 E-钙粘蛋白的表达明显下调。此外,IGF-1 和 IGF-2 的产生明显增加。值得注意的是,外源性 IGF-1/IGF-2 促进了 C643 细胞的侵袭和干性,而阻断 IGF-1 和 IGF-2 通过抑制共培养系统中 IR-A/IGF1R 介导的 PI3K/AKT/mTOR 信号通路抑制转移和干性。IHC 结果表明,CD68 和 CD206 在 ATC 组织中的表达明显增加。综上所述,M2 样 TAMs 加速了 ATC 细胞的转移并增加了其干性,其潜在机制可能与 M2 样 TAMs 对 IGF 的切割有关,该过程激活了 IR-A/IGF1R 介导的 PI3K/AKT/mTOR 信号通路。