PKN2 in colon cancer cells inhibits M2 phenotype polarization of tumor-associated macrophages via regulating DUSP6-Erk1/2 pathway.
PKN2 in colon cancer cells inhibits M2 phenotype polarization of tumor-associated macrophages via regulating DUSP6-Erk1/2 pathway.
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结肠癌细胞中的PKN2通过调节DUSP6-Erk1/2通路抑制肿瘤相关巨噬细胞的M2表型极化
DOI：
10.1186/s12943-017-0747-z
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发表时间：
2018-01-24
期刊：
影响因子：
37.3
通讯作者：
Gong S
中科院分区：
文献类型：
--
作者：
Cheng Y;Zhu Y;Xu J;Yang M;Chen P;Xu W;Zhao J;Geng L;Gong S
研究背景蛋白激酶N2（Protein kinase N2，PKN 2）是一种与PKC相关的丝氨酸/苏氨酸蛋白激酶。PKN 2是肿瘤细胞迁移、侵袭和凋亡所必需的。PKN 2在结肠癌组织中的表达与肿瘤相关巨噬细胞（tumor associated macrophages，TAMs）极化的关系尚不清楚。通过RT-PCR、IHC和流式细胞术评价M1/M2巨噬细胞特征。在体外和体内研究PKN 2对肿瘤生长和TAM极化的影响。PKN 2靶向细胞因子/通路通过基因表达分析进行分析，并通过PCR、荧光素酶测定或蛋白质印迹进行进一步确认。通过ChIP-qPCR证实PKN 2与IL 4和IL 10的转录因子之间的相关性。激酶活性测定法测定PKN 2和DUSP 6的催化活性。PKN 2和DUSP 6之间的相互作用被证实由Co-IP.ResultsThe结肠癌细胞PKN 2的表达预测了良好的预后，并与低M2巨噬细胞在人结肠癌组织中的含量。PKN 2在小鼠异种移植模型中抑制肿瘤生长，并在体外和体内抑制M2表型极化。PKN 2通过抑制Erk 1/2磷酸化来抑制结肠癌细胞中IL 4和IL 10的表达，Erk 1/2磷酸化是CREB和Elk-1磷酸化和结合IL 4和IL 10启动子所必需的。DUSP 6通过与PKN 2直接结合而被磷酸化和激活，抑制Erk 1/2的活化。结论PKN 2在结肠癌细胞中的表达抑制了肿瘤相关的M2巨噬细胞极化和肿瘤生长。针对PKN 2信号通路可能为结肠癌提供一种潜在的治疗策略。
BackgroundProtein kinase N2 (PKN2) is a PKC-related serine/threonine-protein kinase. PKN2 is required for tumor cell migration, invasion and apoptosis. However, the functional role of PKN2 in regulating tumor associated macrophages (TAMs) polarization in colon cancer has never been reported.MethodsPKN2 expression in human colon cancer tissues was examined with immunohistochemistry (IHC). M1/M2 macrophage signatures were evaluated by RT-PCR, IHC and flow cytometry. The effects of PKN2 on tumor growth and TAM polarization were investigated both in vitro and in vivo. PKN2 targeted cytokines/pathway were analyzed by gene expression analysis and further confirmed by PCR, luciferase assay or western blot. Correlations between PKN2 and transcriptional factors for IL4 and IL10 were confirmed by ChIP-qPCR. The catalytic activities of PKN2 and DUSP6 were determined by kinase activity assay. Interactions between PKN2 and DUSP6 were confirmed by Co-IP.ResultsThe expression of PKN2 in colon cancer cells predicted a favorable prognosis and was associated with low M2 macrophage content in human colon cancer tissues. PKN2 inhibited tumor growth in mice xenograft model and inhibited M2 phenotype polarization both in vitro and in vivo. Mechanistically, PKN2 suppresses the expression of IL4 and IL10 from colon cancer cells by inhibiting Erk1/2 phosphorylation, which is required for phosphorylation and binding of CREB and Elk-1 to the promoters of IL4 and IL10. DUSP6, which is phosphorylated and activated through direct association with PKN2, suppresses Erk1/2 activation.ConclusionsThe expression of PKN2 in colon cancer cells suppresses tumor associated M2 macrophage polarization and tumor growth. Targeting PKN2 signaling pathway may provide a potential therapeutic strategy for colon cancer.
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