PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro.
PEDF increases the tumoricidal activity of macrophages towards prostate cancer cells in vitro.
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DOI：
10.1371/journal.pone.0174968
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发表时间：
2017
期刊：
影响因子：
3.7
通讯作者：
Filleur S
中科院分区：
文献类型：
--
作者：
Martinez-Marin D;Jarvis C;Nelius T;de Riese W;Volpert OV;Filleur S
尽管炎症和前列腺癌（PCa）已经联系在一起，但巨噬细胞和前列腺癌细胞之间的分子相互作用尚不清楚。色素上皮衍生因子（PEDF）是一种抗血管生成和抗肿瘤因子。我们之前的研究表明，PEDF在体外诱导巨噬细胞募集，与人前列腺巨噬细胞密度相关，并刺激巨噬细胞向经典激活途径极化。在这里，我们证明PEDF通过双向信号调节巨噬细胞和PCa细胞之间的相互作用，导致肿瘤细胞凋亡和吞噬。RAW 264.7、THP-1细胞和BMDMs分别与PC3或CL1肿瘤细胞单独或共培养。测定PEDF及其衍生的P18肽对巨噬细胞分化、迁移、超氧化物产生以及肿瘤细胞凋亡和吞噬的影响。采用定量RT-PCR、western blot、免疫荧光和流式细胞术检测巨噬细胞和肿瘤细胞中PEDF受体（ATP5B、PNPLA2和LRP6）和CD47 mRNA和蛋白的表达。我们发现PEDF诱导巨噬细胞向肿瘤三维球体和二维培养物迁移。在共培养中，PEDF通过间接凋亡依赖机制增加了PCa细胞的吞噬能力。此外，PEDF刺激巨噬细胞产生超氧化物。与对照条件培养基相比，巨噬细胞暴露于PEDF的条件培养基诱导肿瘤细胞凋亡，表明ROS可能参与肿瘤细胞凋亡。PEDF分别上调和下调巨噬细胞上的ATP5B和PNPLA2 PEDF受体和肿瘤细胞上的CD47受体。作为PEDF，阻断CD47诱导吞噬。抑制ATP5B可减少吞噬作用。相反，PNPLA2抑制抑制分化，但维持吞噬作用。抑制ATP5B可部分恢复cd47诱导的吞噬作用，表明两者具有互补作用。P18 pedf衍生肽也观察到类似的效果。这些数据表明，使用PEDF调节巨噬细胞与PCa细胞之间的分子相互作用可能是治疗PCa的一种有希望的策略。
Although inflammation and prostate cancer (PCa) have been linked, the molecular interactions between macrophages and PCa cells are poorly explored. Pigment Epithelium-Derived Factor (PEDF) is an anti-angiogenic and anti-tumor factor. We previously showed that PEDF induces macrophages recruitment in vitro, correlates with macrophages density in human prostate, and stimulates macrophages polarization towards the classically activated pathway. Here, we demonstrate that PEDF modulates the interaction between macrophages and PCa cells through a bidirectional signalling leading to tumor cell apoptosis and phagocytosis. RAW 264.7 and THP-1 cells, and BMDMs were grown in vitro as mono- or co-cultures with PC3 or CL1 tumor cells. The effects of PEDF and its derived P18 peptide were measured on macrophages differentiation, migration, and superoxide production, and tumor cell apoptosis and phagocytosis. PEDF receptors (ATP5B, PNPLA2, and LRP6) and CD47 mRNA and protein expression were quantified in macrophages and tumor cells by quantitative RT-PCR, western blot, immunofluorescence and flow cytometry. We found that PEDF induced the migration of macrophages towards tumor 3D spheroids and 2D cultures. In co-culture, PEDF increased PCa cells phagocytosis through an indirect apoptosis-dependent mechanism. Moreover, PEDF stimulated the production of superoxide by macrophages. Conditioned media from macrophages exposed to PEDF induced tumor cells apoptosis in contrast to control conditioned media suggesting that ROS may be involved in tumor cells apoptosis. ATP5B and PNPLA2 PEDF receptors on macrophages and CD47 on tumor cells were respectively up- and down-regulated by PEDF. As PEDF, blocking CD47 induced phagocytosis. Inhibiting ATP5B reduced phagocytosis. Inversely, PNPLA2 inhibition blocks differentiation but maintains phagocytosis. CD47-induced phagocytosis was partially reverted by ATP5B inhibition suggesting a complementary action. Similar effects were observed with P18 PEDF-derived peptide. These data established that modulating the molecular interactions between macrophages and PCa cells using PEDF may be a promising strategy for PCa treatment.