Laboratoire M,E,R,C,I - EA 3829, Faculté de Médecine et de Pharmacie, Université de Rouen, 76183 Rouen cedex, France.
BMC Cancer. 2010 Jul 17;10:375. doi: 10.1186/1471-2407-10-375.
Infiltration by macrophages (Mphi) indicates a poor prognosis in breast cancers, in particular by inducing angiogenesis. Our study aimed 1) to investigate the mechanism by which cooperation between Mphi and aggressive breast cancer cells (MDA-MB-231) induces angiogenesis; 2) to examine the effect of tetrathiomolybdate (TM) on this angiogenic activity.
Mphi coincubated with MDA-MB-231 were used as a model to mimic the inflammatory microenvironment. Angiogenesis induced by the culture media was tested in the chick chorioallantoic membrane (CAM). Mphi phenotype was evaluated by 1) expression of the M1 marker CD80, and secretion of interleukin 10 (IL-10), an M2 marker; 2) capacity to secrete Tumour Necrosis Factor alpha (TNFalpha) when stimulated by lipopolysaccharide/interferon gamma (LPS/IFNgamma); 3) ability to induce MDA-MB-231 apoptosis. To explore the molecular mechanisms involved, cytokine profiles of conditioned media from MDA-MB-231, Mphi and the coculture were characterised by an antibody cytokine array. All experiments were carried out both in presence and in absence of TM.
Incubation of Mphi with MDA-MB-231 induced a pro-angiogenic effect in the CAM. It emerged that the angiogenic activity of the coculture is due to the capacity of Mphi to switch from M1 Mphi towards M2, probably due to an increase in Macrophage Colony Stimulating Factor. This M1-M2 switch was shown by a decreased expression of CD80 upon LPS/IFNgamma stimulation, an increased secretion of IL-10, a decreased secretion of TNFalpha in response to LPS/IFNgamma and an inability to potentiate apoptosis. At the molecular level, the angiogenic activity of the coculture medium can be explained by the secretion of CXC chemokines/ELR+ and CC chemokines. Although TM did not modify either the M2 phenotype in the coculture or the profile of the secreted chemokines, it did decrease the angiogenic activity of the coculture medium, suggesting that TM inhibited angiogenic activity by interfering with the endothelial cell signalling induced by these chemokines.
Cooperation between Mphi and MDA-MB-231 transformed M1 Mphi to an angiogenic, M2 phenotype, attested by secretion of CXC chemokines/ELR+ and CC chemokines. TM inhibited this coculture-induced increase in angiogenic activity, without affecting either Mphi phenotype or cytokine secretion profiles.
巨噬细胞（Mphi）浸润表明乳腺癌预后不良，特别是通过诱导血管生成。我们的研究旨在 1）研究巨噬细胞与侵袭性乳腺癌细胞（MDA-MB-231）合作诱导血管生成的机制；2）研究四硫钼酸盐（TM）对这种血管生成活性的影响。
用巨噬细胞与 MDA-MB-231 共培养作为模型来模拟炎症微环境。用鸡胚绒毛尿囊膜（CAM）检测培养基诱导的血管生成。通过 1）M1 标志物 CD80 的表达和 M2 标志物白细胞介素 10（IL-10）的分泌，评估巨噬细胞表型；2）用脂多糖/干扰素γ（LPS/IFNγ）刺激时分泌肿瘤坏死因子α（TNFalpha）的能力；3）诱导 MDA-MB-231 凋亡的能力。为了探索所涉及的分子机制，用抗体细胞因子阵列对 MDA-MB-231、巨噬细胞和共培养物的条件培养基中的细胞因子谱进行了表征。所有实验均在有和没有 TM 的情况下进行。
巨噬细胞与 MDA-MB-231 孵育在 CAM 中诱导了促血管生成作用。结果表明，共培养的血管生成活性归因于巨噬细胞从 M1 巨噬细胞向 M2 巨噬细胞的转变能力，这可能是由于巨噬细胞集落刺激因子的增加。这种 M1-M2 转换表现为 LPS/IFNγ 刺激时 CD80 的表达降低，IL-10 分泌增加，LPS/IFNγ 反应时 TNFalpha 分泌减少，以及不能增强细胞凋亡。在分子水平上，共培养培养基的血管生成活性可以用 CXC 趋化因子/ELR+和 CC 趋化因子的分泌来解释。尽管 TM 既不改变共培养物中的 M2 表型，也不改变分泌的趋化因子谱，但它确实降低了共培养物培养基的血管生成活性，这表明 TM 通过干扰这些趋化因子诱导的内皮细胞信号转导来抑制血管生成活性。
巨噬细胞与 MDA-MB-231 的合作将 M1 巨噬细胞转化为血管生成的 M2 表型，这表现在 CXC 趋化因子/ELR+和 CC 趋化因子的分泌上。TM 抑制了这种共培养诱导的血管生成活性增加，而不影响巨噬细胞表型或细胞因子分泌谱。
