Department of Immunology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; Cancer, Petroleum and Environmental Pollutants Research Center, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
Department of Immunology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran; Cellular and Molecular Research Center, Medical Basic Sciences Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
Int Immunopharmacol. 2024 May 10;132:111960. doi: 10.1016/j.intimp.2024.111960. Epub 2024 Mar 29.
Scorpion venoms identified as agents with anti-tumor and anti-angiogenic features. Tumor microenvironment (TME) plays a pivotal role in the process of tumorigenesis, tumor development, and polarization of M2 phenotype tumor associated macrophages (TAMs). M2 polarized cells are associated with tumor growth, invasion, and metastasis. The fractionation process was performed by gel filtration chromatography on a Sephadex G50 column. To elucidate whether scorpion venom can alter macrophage polarization, we treated interleukin (IL)-4-polarized M2 cells with isolated fractions from Mesobuthus eupeus. Next, we evaluated the cytokine production and specific markers expression for M2 and M1 phenotype using enzyme linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (PCR), respectively. The phagocytic capacity of macrophages was also assessed. In addition, the migration assay and MTT analysis were performed to investigate the effects of reprogrammed macrophages on the CT-26 colon cancer cells. The results indicated that F1 fraction of venom significantly upregulated the levels and expression of M1-associated cytokines and markers, including tumor necrosis factor-alpha (TNF-α) (p < 0.001), IL-1 (p < 0.01), interferon regulatory factor 5 (IRF5) (p < 0.0001), induced nitric oxide synthase (iNOS) (p < 0.0001), and CD86 (p < 0.0001), and downregulated M2-related markers, including transforming growth factor-beta (TGF-β) (p < 0.05), IL-10 (p < 0.05), Fizz1 (p < 0.0001), arginase-1 (Arg-1) (p < 0.0001), and CD206 (p < 0.001). The macrophage phagocytic capacity was enhanced after treatment with F1 fraction (p < 0.01). Moreover, incubation of CT-26 cell line with conditioned media of F1-treated macrophages suppressed migration (p < 0.0001) and proliferation (p < 0.01) of tumor cells. In conclusion, our findings demonstrated the potential of Mesobuthus eupeus venom in M2-to-M1 macrophage polarization as a promising therapeutic approach against proliferation and metastasis of colon cancer cells.
蝎毒液被鉴定为具有抗肿瘤和抗血管生成作用的药物。肿瘤微环境（TME）在肿瘤发生、肿瘤发展和 M2 表型肿瘤相关巨噬细胞（TAMs）极化过程中起着关键作用。M2 极化细胞与肿瘤生长、浸润和转移有关。通过 Sephadex G50 柱凝胶过滤层析进行分级分离过程。为了阐明蝎毒液是否可以改变巨噬细胞极化，我们用从 Mesobuthus eupeus 中分离的各馏分处理白细胞介素（IL）-4 诱导的 M2 细胞。接下来，我们使用酶联免疫吸附测定（ELISA）和实时聚合酶链反应（PCR）分别评估 M2 和 M1 表型的细胞因子产生和特异性标志物表达。还评估了巨噬细胞的吞噬能力。此外，还进行了迁移实验和 MTT 分析，以研究重编程巨噬细胞对 CT-26 结肠癌细胞的影响。结果表明，毒液的 F1 馏分显著上调了 M1 相关细胞因子和标志物的水平和表达，包括肿瘤坏死因子-α（TNF-α）（p < 0.001）、白细胞介素-1（IL-1）（p < 0.01）、干扰素调节因子 5（IRF5）（p < 0.0001）、诱导型一氧化氮合酶（iNOS）（p < 0.0001）和 CD86（p < 0.0001），并下调了 M2 相关标志物，包括转化生长因子-β（TGF-β）（p < 0.05）、白细胞介素-10（IL-10）（p < 0.05）、Fizz1（p < 0.0001）、精氨酸酶-1（Arg-1）（p < 0.0001）和 CD206（p < 0.001）。用 F1 馏分处理后，巨噬细胞的吞噬能力增强（p < 0.01）。此外，用 F1 处理的巨噬细胞条件培养基孵育 CT-26 细胞系可抑制肿瘤细胞的迁移（p < 0.0001）和增殖（p < 0.01）。总之，我们的研究结果表明，Mesobuthus eupeus 毒液具有将 M2 向 M1 巨噬细胞极化的潜力，是一种有前途的治疗结肠癌增殖和转移的方法。
