Inhibition of let-7b-5p contributes to an anti-tumorigenic macrophage phenotype through the SOCS1/STAT pathway in prostate cancer.
Inhibition of let-7b-5p contributes to an anti-tumorigenic macrophage phenotype through the SOCS1/STAT pathway in prostate cancer.
复制标题
DOI：
10.1186/s12935-020-01563-7
复制
发表时间：
2020
影响因子：
5.8
通讯作者：
Wang Z
中科院分区：
文献类型：
--
作者：
Rong J;Xu L;Hu Y;Liu F;Yu Y;Guo H;Ni X;Huang Y;Zhao L;Wang Z
microRNAs（miRNAs）功能障碍是炎症细胞因子异常表达的主要原因，并有助于巨噬细胞极化。促炎性M1巨噬细胞促进辅助性T细胞（Th）1应答并显示出杀肿瘤活性，而M2巨噬细胞在组织修复和重塑中显示出调节功能并促进Th 2免疫应答。先前的研究表明，miRNA let-7与细胞分化相关，并且let-7 b-5 p的表达在M2巨噬细胞中显著增强。然而，let-7 b-5 p调节前列腺癌（PCa）中巨噬细胞分化的机制在很大程度上仍然未知。通过来自健康男性供体的血液单核细胞诱导人巨噬细胞，并通过用100 ng/ml脂多糖和100 ng/ml IFN-γ刺激M1巨噬细胞过夜使其极化。用PC-3细胞的条件培养基体外诱导前列腺巨噬细胞（M-CM），然后将let-7 b-5 p模拟物或抑制剂转染M1和M-CM 72 h。采用高分辨率内涵式图像分析系统检测各组细胞分化抗原206（CD 206）的表达。我们使用定量实时聚合酶链反应（qRT-PCR）检测巨噬细胞中炎性细胞因子IL-10、IL-12、IL-13、TNF-α和let-7 b的表达。ELISA法检测SOCS 1蛋白水平，CST信号通路芯片检测STAT家族成员蛋白磷酸化差异。采用吞噬实验、CCK-8细胞计数试剂盒和集落形成实验检测巨噬细胞的吞噬功能和巨噬细胞对前列腺癌PC-3细胞增殖的影响。用双荧光素酶报告基因证实了SOCS 1和let-7 b-5 p之间的关系。在用let-7 b-5 p模拟物处理的M1巨噬细胞中，分化簇206（CD 206，M2样巨噬细胞表面分子）的表达显著增加，而在用let-7 b-5 p抑制剂处理的M-CM中，CD 206表达降低。let-7 b-5 p的过表达或敲低显著影响巨噬细胞中炎症因子的表达，包括白细胞介素10（IL-10）、IL-12、IL-13和肿瘤坏死因子α。与其他组相比，let-7 b-5 p在M-CM和M1巨噬细胞中下调细胞因子信号转导抑制因子1（SOCS 1）的表达，并增加信号转导和转录激活因子1（STAT 1），STAT 3和STAT 5a蛋白的磷酸化。此外，随着let-7 b-5 p表达的增加，巨噬细胞的吞噬作用表现出相应的显著降低。结果，用let-7 b-5 p抑制剂处理的M-CM减少PC-3 PCa细胞的增殖。总的来说，这些数据表明let-7 b-5 p可以通过SOCS 1/STAT途径调节M2极化，并且let-7 b-5 p抑制剂逆转M2分化增强巨噬细胞吞噬作用，最终抑制PCa细胞的增殖。
Dysfunction of microRNAs (miRNAs) is a major cause of aberrant expression of inflammatory cytokines and contributes to macrophage polarization. Proinflammatory M1 macrophages promote T helper (Th) 1 responses and show tumoricidal activity, whereas M2 macrophages display regulatory functions in tissue repair and remodeling and promote Th2 immune responses. Previous studies have shown that miRNA let-7 is associated with cellular differentiation and that the expression of let-7b-5p is significantly augmented in M2 macrophages. However, the mechanism by which let-7b-5p regulates macrophage differentiation in prostate cancer (PCa) remains largely unknown. Human macrophages were induced by blood monocytes from healthy male donors, and M1 macrophages were polarized by stimulating them overnight with 100 ng/ml of lipopolysaccharides and 100 ng/ml of IFN-γ. Conditioned medium from PC-3 cells was used to induce prostatic macrophages (M-CMs) in vitro, and we then transfected let-7b-5p mimics or inhibitors into M1 and M-CMs for 72 h. The expression of cluster of differentiation 206 (CD206) in each group was detected with the High-Throughput Connotation of Imaging System. We used quantitative real-time polymerase chain reaction (qRT-PCR) to examine the expression of the inflammatory cytokines IL-10, IL-12, IL-13, TNF-alpha, and let-7b in macrophages. SOCS1 protein levels were evaluated by ELISA, and the phosphorylation difference in STAT family member proteins was analyzed using CST signal-pathway chip. Phagocytosis by macrophages and the effect of macrophages on the proliferation of prostate cancer PC-3 cells were evaluated with phagocytosis assay or the Cell Counting Kit-8 (CCK-8) and colony formation assay. The relationship between SOCS1 and let-7b-5p was confirmed with a dual-luciferase reporter. The expression of cluster of differentiation 206 (CD206, a M2-like macrophage surface molecule) was significantly increased in M1 macrophages treated with let-7b-5p mimics, while CD206 expression was decreased in M-CMs treated with let-7b-5p inhibitors. Overexpression or knockdown of let-7b-5p significantly affected the expression of inflammatory factors in macrophages—including interleukin 10 (IL-10), IL-12, IL-13, and tumor necrosis factor alpha. Let-7b-5p downregulated the expression of suppressor of cytokine signaling 1 (SOCS1) and increased the phosphorylation of signal transducer and activator of transcription 1 (STAT1), STAT3, and STAT5a proteins in M-CMs and M1 macrophages with let-7b-5p mimics relative to the other groups. In addition, with the elevated expression of let-7b-5p, the phagocytosis by macrophages showed a commensurate and significant decrease. As a result, M-CMs treated with let-7b-5p inhibitors reduced the proliferation of PC-3 PCa cells. Collectively, these data indicated that let-7b-5p may regulate M2 polarization through the SOCS1/STAT pathway and that reversal of M2 differentiation by let-7b-5p inhibitors enhanced macrophage phagocytosis, ultimately inhibiting the proliferation of PCa cells.
登录
查看更多内容
DOI：
10.4049/jimmunol.1202496
发表时间：
2013-06-15
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Banerjee S;Xie N;Cui H;Tan Z;Yang S;Icyuz M;Abraham E;Liu G
通讯作者：
Liu G
影响因子：
4.8
作者：
McCormick, Sarah M.;Gowda, Nagaraj;Heller, Nicola M.
通讯作者：
Heller, Nicola M.
影响因子：
82.9
作者：
Moore, RJ;Owens, DM;Balkwill, F
通讯作者：
Balkwill, F
影响因子：
6.6
作者：
Jacobsson, Hanna;Harrison, Hannah;Landberg, Goran
通讯作者：
Landberg, Goran
影响因子：
20.3
作者：
Lu, Changming;Huang, Xin;Zhou, Xianzheng
通讯作者：
Zhou, Xianzheng