替代激活的单核细胞促进人乳腺癌细胞SKBR3的生长侵袭

Alternatively activated macrophages /mononuclear phagocytes promote growth and invasion of breast cancer cell line SKBR3

目的研究不同激活状态的单核细胞在乳腺癌发展中的作用,探索肿瘤相关巨噬细胞在乳腺癌中的作用机制。方法体外分离培养人外周血单核细胞,分为经典激活、替代激活、空白对照3组,分别采用细菌脂多糖IL-4、培养基进行相应激活。48～72h后,采用RT-PCR、与人乳腺癌细胞株SKBR3共培养、建立鸡胚尿膜囊血管生成模型等方法检测各组TNF-α、AMAC-1、β-actinmRNA表达的差异,观察3种不同活化状态的单核巨噬细胞对乳腺癌细胞生长的影响以及对肿瘤诱导新生血管形成的影响。结果经典激活组高表达TNF-α,替代激活组则高表达AMAC-1,3组间分子标志物的表达差异具有统计学意义(P<0.05)。共培养实验显示经典激活的单核细胞呈现出明显抑瘤效应,3d后效应明显高于其他两组(P<0.05),替代激活的单核细胞则在第5天开始呈现出明显促肿瘤效应(P<0.05)。在鸡胚尿囊膜血管生成模型中,替代激活的单核细胞可以明显促进肿瘤新生血管的生成(57.33±14.82),与经典激活(20.44±6.50)及未激活组(32.22±8.33)相比,差异有统计学意义(P<0.05)。结论IL-4可以促进人外周血单核细胞向替代激活的方向分化并高表达替代激活的分子标志物AMAC-1,而替代激活的单核巨噬细胞在与肿瘤细胞的共培养中可以促进肿瘤细胞的生长以及刺激肿瘤新生血管的生成。提示肿瘤相关巨噬细胞在体内很可能主要以替代激活的方式存在,从而直接或间接地促进了肿瘤的进展。

Objective To study the effect of altematively activated macrophages/mononuclear phagocytes （MNP） on breast cancer cells and explore the mechanisms for the action of tumor-associated macrophages in breast cancer. Methods Human peripheral blood monocytes were isolated and cultured in vitro and divided into 3 groups, namely classically activated monocytes （CAM） which were induced by lipopolysaccharide, altematively activated monocytes （AAM） induce by IL-4, and control cells treated with the culture medium only. After cell culture for 48-72 h, the mRNA of tumor necrosis factor-α （TNF-α, altemative monocytes activation- associated CC-chemokine 1 （AMAC-1）, and β-actin of the 3 groups were extracted for P,T-PCR, or the cells were cocultured with breast cancer cell line SKBP,3, or seeded in chicken chorioallantoic membrane along with SKBR3. Results TNF-α mRNA was significantly increased in CAM, and AMAC-1 was highly expressed in AAM. The coculture experiments showed that CAM exhibited obvious inhibitory effect on SKBR3 cells after a 3-day culture whereas AAM significantly promoted the growth of SKBR3 cells after a 5-day culture. In chioken on chorioallantoic membrane experiment, the macrophages promoted tumor angiogenesis and AAM showed the most obvious effect. Conclusion IL-4 induces high expression of AMAC- 1, a molecular marker of AAM, in the macrophages, and AAM can promote the growth of SKBR3 cells and tumor angiogenesis.