目的:骨肉瘤是一种缺乏最佳临床治疗方案的原发性骨肿瘤。肿瘤微环境中的肿瘤相关巨噬细胞与肿瘤的发展和转移密切相关。研究已经确定具有胶原结构的巨噬细胞受体(MARCO)是在巨噬细胞中表达的特异性受体。本研究旨在研究抗MARCOmAb治疗是否可以诱导肿瘤微环境中的巨噬细胞极化并引起抗肿瘤作用。
方法:THP-1细胞用20ng/mL佛波醇12-肉豆蔻酸酯13-乙酸酯和80ng/mL白介素-4处理48小时,以诱导巨噬细胞极化为交替活化的巨噬细胞(M2)。酶联免疫吸附测定,实时定量聚合酶链反应,流式细胞术,和生物信息学分析进行评估巨噬细胞极化。共培养组包括一个空白组,M2巨噬细胞和U2OS共培养组,和抗MARCOmAb处理的M2巨噬细胞组。细胞活力测定,细胞划痕测试,凋亡,进行细胞周期分析以确定抗MARCOmAb处理的巨噬细胞对骨肉瘤细胞的作用。
结果:证明了抗MARCOmAb可以驱动巨噬细胞向经典激活的巨噬细胞(M1)极化。抗MARCO单克隆抗体促进巨噬细胞分泌促炎因子,包括肿瘤坏死因子-α(TNF-α),白细胞介素-1β,白细胞介素-6和白细胞介素-23.对体外共培养模型的研究表明,用抗MARCOmAb处理的巨噬细胞可以抑制骨肉瘤细胞的生长和迁移,诱导细胞凋亡,并通过巨噬细胞的M1极化抑制骨肉瘤细胞的细胞周期进程。
结论:抗MARCOmAb治疗通过影响巨噬细胞向M1巨噬细胞的极化发挥抗骨肉瘤作用,为治疗骨肉瘤提供了一种潜在的新治疗方法。
OBJECTIVE: Osteosarcoma is a primary bone tumor lacking optimal clinical treatment options. Tumor-associated macrophages in the tumor microenvironment are closely associated with tumor development and metastasis. Studies have identified the macrophage receptor with collagenous structure (MARCO) as a specific receptor expressed in macrophages. This study aimed to investigate whether anti-MARCO mAb treatment can induce macrophage polarization in the tumor microenvironment and elicit anti-tumor effects.
METHODS: THP-1 cells were treated with 20 ng/mL phorbol 12-myristate 13-acetate and 80 ng/mL interleukin-4 for 48 h to induce macrophage polarization to alternatively activated macrophages (M2). Enzyme-linked immunosorbent assay, real-time quantitative polymerase chain reaction, flow cytometry, and bioinformatic analyses were performed to evaluate macrophage polarization. The co-culture groups included a blank group, an M2 macrophage and U2OS co-culture group, and an anti-MARCO mAb-treated M2 macrophage group. Cell viability assays, cell scratch tests, apoptosis, and cell cycle analyses were performed to determine the effects of anti-MARCO mAb-treated macrophages on osteosarcoma cells.
RESULTS: It was demonstrated that anti-MARCO mAb can drive macrophages toward classically activated macrophage (M1) polarization. Anti-MARCO mAb promoted the secretion of pro-inflammatory factors by macrophages, including tumor necrosis factor-alpha (TNF-α), interleukin-1beta, interleukin-6 and interleukin-23. Studies on in vitro co-culture models have revealed that macrophages treated with anti-MARCO mAb can suppress the growth and migration of osteosarcoma cells, induce cell apoptosis, and inhibit cell cycle progression of osteosarcoma cells through M1 polarization of macrophages in vitro.
CONCLUSIONS: Anti-MARCO mAb treatment exerts anti-osteosarcoma effects by affecting macrophage polarization toward M1 macrophages, offering a potential new therapeutic approach for treating osteosarcoma.