Department of Hepatobiliary Surgery, Tianjin First Central Hospital, School of Medicine, Nankai University, Tianjin, China.
School of Medicine, Nankai University, Tianjin, China.
Front Immunol. 2023 Sep 22;14:1251648. doi: 10.3389/fimmu.2023.1251648. eCollection 2023.
The relationship between the tumor microenvironment and the network of key signaling pathways in cancer plays a key role in the occurrence and development of tumors. Tumor-associated macrophages (TAMs) are important inflammatory cells in the tumor microenvironment and play an important role in tumorigenesis and progression. Macrophages in malignant tumors, mainly the M2 subtype, promote tumor progression by producing cytokines and down-regulating anti-inflammatory immune responses. Several articles have investigated the effect of macrophages on the sensitivity of cancer chemotherapeutic agents, but few such articles have been reported in cholangiocarcinoma, so we investigated the effect of M2 macrophage on the sensitivity of cholangiocarcinoma cells to Lenvatinib compared to M1.
THP-1 monocytes were polarized to M0 macrophage by phorbol 12-myristate 13-acetate (PMA) and then induced to differentiate into M1 and M2 macrophages by LPS, IFN-γ and IL-4 and IL-13, respectively. Macrophages and cholangiocarcinoma cells were co-cultured prior to 24 hours of Lenvatinib administration, cancer cell apoptosis was detected by western-blot, FACS analysis of Annexin V and PI staining. Furthermore, we use xCELLigence RTCA SP Instrument (ACEA Bio-sciences) to monitor cell viability of Lenvatinib administration in co-culture of cholangiocarcinoma cells and macrophages. After tumorigenesis in immunodeficient mice, Lenvatinib was administered, and the effects of M2 on biological characteristics of cholangiocarcinoma cells were investigated by immuno-histochemistry.
mRNA and protein expression of M1 and M2 markers confirmed the polarization of THP-1 derived macrophages, which provided a successful and efficient model of monocyte polarization to TAMs. Lenvatinib-induced apoptosis of cholangiocarcinoma cells was significantly reduced when co-cultured with M2 macrophage, whereas apoptosis of cholangiocarcinoma cells co-cultured with M1 macrophage was increased. In the CDX model, Lenvatinib-induced cancer cell apoptosis was markedly reduced, and proliferative cells increased in the presence of M2 macrophages. Angiogenesis related factors was significantly increased in cholangiocarcinoma cells co-cultured with M2.
Compared with M1, M2 macrophages can inhibit the anti-tumor effect of Lenvatinib on cholangiocarcinoma through immune regulation, which may be related to the tumor angiogenesis factor effect of M2 macrophage.
肿瘤微环境与癌症关键信号通路网络之间的关系在肿瘤的发生和发展中起着关键作用。肿瘤相关巨噬细胞（TAMs）是肿瘤微环境中的重要炎性细胞，在肿瘤发生和进展中发挥重要作用。恶性肿瘤中的巨噬细胞，主要是 M2 亚型，通过产生细胞因子和下调抗炎免疫反应来促进肿瘤进展。有几篇文章研究了巨噬细胞对癌症化疗药物敏感性的影响，但在胆管癌中很少有此类文章报道，因此我们研究了与 M1 相比，M2 巨噬细胞对胆管癌细胞对仑伐替尼敏感性的影响。
用佛波醇 12-肉豆蔻酸 13-乙酸酯（PMA）将 THP-1 单核细胞极化为 M0 巨噬细胞，然后用 LPS、IFN-γ 和 IL-4、IL-13 分别诱导分化为 M1 和 M2 巨噬细胞。在给予仑伐替尼前 24 小时，将巨噬细胞和胆管癌细胞共培养，通过 Western blot、Annexin V 和 PI 染色的 FACS 分析检测癌细胞凋亡。此外，我们使用 xCELLigence RTCA SP 仪器（ACEA Bio-sciences）监测共培养的胆管癌细胞和巨噬细胞中仑伐替尼给药后的细胞活力。在免疫缺陷小鼠肿瘤发生后，给予仑伐替尼，并通过免疫组织化学研究 M2 对胆管癌细胞生物学特性的影响。
M1 和 M2 标志物的 mRNA 和蛋白表达证实了 THP-1 衍生巨噬细胞的极化，这为单核细胞向 TAMs 极化为成功和有效的模型提供了依据。当与 M2 巨噬细胞共培养时，仑伐替尼诱导的胆管癌细胞凋亡明显减少，而与 M1 巨噬细胞共培养时胆管癌细胞凋亡增加。在 CDX 模型中，仑伐替尼诱导的癌细胞凋亡明显减少，而存在 M2 巨噬细胞时增殖细胞增加。与 M2 共培养的胆管癌细胞中血管生成相关因子显著增加。
与 M1 相比，M2 巨噬细胞可通过免疫调节抑制仑伐替尼对胆管癌的抗肿瘤作用，这可能与 M2 巨噬细胞的肿瘤血管生成因子作用有关。
