目的:M2样肿瘤相关巨噬细胞(TAMs)在促进肿瘤增殖中起关键作用,血管生成,和转移。在目前的研究中,我们研究了巨噬细胞极化与阿曲苯内酯II(AT-II)在肺癌细胞中的抗肿瘤作用之间的关系。
方法:细胞活力,迁移,用MTT法测定侵袭,伤口愈合试验,和transwell分析,分别。流式细胞术分析显示CD206+细胞的百分比。通过实时PCR测定基因表达,西方印迹,和免疫荧光染色。使用Lewis肺癌小鼠异种移植和转移模型来检查AT-II对体内肺癌的影响。
结果:AT-II(2.5和5μM)未引起A549细胞活力的显著抑制,但显著抑制IL-4/IL-13诱导的M2样极化,M2表面标记CD206的表达降低,特异性M2标记基因(Arg-1,IL-10和TGF-β)的下调以及M2巨噬细胞介导的A549细胞的侵袭和迁移的抑制。此外,AT-II抑制IL-4/IL-13诱导的STAT6信号传导途径的激活,这在巨噬细胞的M2样极化中是至关重要的。在动物模型中,AT-II的给药(50mgkg-1,i.g.,QD21天)显着抑制肿瘤生长,肺转移结节减少,并下调肿瘤组织和肺转移结节中M2巨噬细胞(F4/80+和CD206+)在总巨噬细胞(F4/80+)中的百分比。
结论:AT-II有效抑制M2样极化,从而在体内和体外抑制肺癌细胞转移,揭示了AT-II抗肿瘤作用的新潜在策略。
OBJECTIVE: M2-like tumor-associated macrophages (TAMs) play a crucial role in promoting tumor proliferation, angiogenesis, and metastasis. In the current study, we investigated the relationship between macrophage polarization and the antitumor effect of Atractylenolide II (AT-II) in lung cancer cells.
METHODS: Cell viability, migration, and invasion were determined by MTT assay, wound healing assay, and transwell assay, respectively. Flow cytometry analysis showed the percentage of CD206+ cells. Gene expression was determined by real-time PCR, western blotting, and immunofluorescence staining. Lewis lung carcinoma mouse xenograft and metastasis models were used to examine the effects of AT-II on lung cancer in vivo.
RESULTS: AT-II (2.5 and 5 µM) did not cause significant inhibition of A549 cell viability but markedly inhibited IL-4/IL-13-induced M2-like polarization, evidenced by the decreased expression of the M2 surface marker CD206, down-regulation of specific M2-marker genes (Arg-1, IL-10 and TGF-β) as well as inhibition of M2 macrophages-mediated invasion and migration of A549 cells. In addition, AT-II inhibited IL-4/IL-13-induced activation of the STAT6 signaling pathway that is vital in the M2-like polarization of macrophages. In animal models, administration of AT-II (50 mg kg-1, i.g., QD for 21 days) significantly inhibited tumor growth, reduced pulmonary metastatic nodules, and down-regulated the percentages of M2 macrophages (F4/80+ and CD206+) in total macrophages (F4/80+) in tumor tissues and pulmonary metastatic nodules.
CONCLUSIONS: AT-II effectively inhibits M2-like polarization, thereby inhibiting lung cancer cell metastasis both in vivo and in vitro, revealing a novel potential strategy for the antitumor effect of AT-II.