M2 macrophages induce ovarian cancer cell proliferation via a heparin binding epidermal growth factor/matrix metalloproteinase 9 intercellular feedback loop.
M2 macrophages induce ovarian cancer cell proliferation via a heparin binding epidermal growth factor/matrix metalloproteinase 9 intercellular feedback loop.
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DOI：
10.18632/oncotarget.13474
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发表时间：
2016-12-27
期刊：
影响因子：
--
通讯作者：
Kreeger PK
中科院分区：
文献类型：
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作者：
Carroll MJ;Kapur A;Felder M;Patankar MS;Kreeger PK
在卵巢癌中，抗炎M2与促炎M1巨噬细胞比例高与患者预后不良相关。由于M2巨噬细胞在肿瘤微环境中的存在而导致肿瘤预后不良的机制尚不清楚，并且用现有技术进行研究具有挑战性。因此，在本研究中，我们利用我们实验室先前开发的微培养装置来模拟集中的旁分泌信号，以解决我们的假设，即M2巨噬细胞和卵巢癌细胞之间的相互作用诱导肿瘤细胞增殖。通过微培养装置，我们确定与m2分化的原代巨噬细胞或THP-1共培养可使OVCA433的增殖增加10-12%。表皮生长因子受体（EGFR）或肝素结合表皮生长因子（HB-EGF）中和抗体消除了这种影响，卵巢癌患者外周血单个核细胞中HBEGF的表达比健康个体高9倍，表明HB-EGF在肿瘤进展中起作用。然而，在巨噬细胞或巨噬细胞条件培养基分泌的水平上添加HB-EGF并没有达到同样程度的诱导增殖，这表明在这一过程中有其他因素的作用。基质金属蛋白酶-9 （Matrix metalloproteinase-9, MMP-9）可切割膜结合的HB-EGF，在共培养中升高，其抑制作用降低了增殖。在每个群体中利用抑制剂和siRNA对抗MMP9，我们确定巨噬细胞分泌的MMP-9从巨噬细胞中释放HB-EGF，从而增加OVCA433中的MMP9，从而形成一个正反馈循环，驱动HB-EGF释放并增加共培养中的增殖。诸如此类的多细胞相互作用的鉴定可能为如何最有效地控制卵巢癌的进展提供见解。
In ovarian cancer, a high ratio of anti-inflammatory M2 to pro-inflammatory M1 macrophages correlates with poor patient prognosis. The mechanisms driving poor tumor outcome as a result of the presence of M2 macrophages in the tumor microenvironment remain unclear and are challenging to study with current techniques. Therefore, in this study we utilized a micro-culture device previously developed by our lab to model concentrated paracrine signaling in order to address our hypothesis that interactions between M2 macrophages and ovarian cancer cells induce tumor cell proliferation. Using the micro-culture device, we determined that co-culture with M2-differentiated primary macrophages or THP-1 increased OVCA433 proliferation by 10–12%. This effect was eliminated with epidermal growth factor receptor (EGFR) or heparin-bound epidermal growth factor (HB-EGF) neutralizing antibodies and HBEGF expression in peripheral blood mononuclear cells from ovarian cancer patients was 9-fold higher than healthy individuals, suggesting a role for HB-EGF in tumor progression. However, addition of HB-EGF at levels secreted by macrophages or macrophage-conditioned media did not induce proliferation to the same extent, indicating a role for other factors in this process. Matrix metalloproteinase-9, MMP-9, which cleaves membrane-bound HB-EGF, was elevated in co-culture and its inhibition decreased proliferation. Utilizing inhibitors and siRNA against MMP9 in each population, we determined that macrophage-secreted MMP-9 released HB-EGF from macrophages, which increased MMP9 in OVCA433, resulting in a positive feedback loop to drive HB-EGF release and increase proliferation in co-culture. Identification of multi-cellular interactions such as this may provide insight into how to most effectively control ovarian cancer progression.