Wnt/β-Catenin Signaling Regulates Cytokine-Induced Human Inducible Nitric Oxide Synthase Expression by Inhibiting Nuclear Factor-κB Activation in Cancer Cells
Wnt/β-Catenin Signaling Regulates Cytokine-Induced Human Inducible Nitric Oxide Synthase Expression by Inhibiting Nuclear Factor-κB Activation in Cancer Cells
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DOI：
10.1158/0008-5472.can-09-0014
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发表时间：
2009-05-01
期刊：
影响因子：
11.2
通讯作者：
Geller, David A.
中科院分区：
文献类型：
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作者：
Du, Qiang;Zhang, Xinglu;Geller, David A.
人诱导型一氧化氮合酶（hiNOS）基因受核因子κ B（NF-κ B）调节，并且最近已被证明是Wnt/β-连环蛋白途径的靶点。在这项研究中，我们验证了Wnt/β-连环蛋白信号可能通过与NF-κ B相互作用调节细胞因子或肿瘤坏死因子α（TNF α）诱导的hiNOS表达的假设。TNF α +白细胞介素（IL）-1 β + IFN γ的细胞因子混合物诱导HCT 116和DLD 1结肠癌细胞中hiNOS启动子活性增加2至3倍，但在SW 480结肠癌细胞中产生2倍的降低。在肝癌细胞（HepG 2、Huh 7和Hep 3B）中观察到类似的差异活性。β-连环蛋白的过表达导致NF-κ B报告基因活性呈剂量依赖性降低，并降低细胞因子混合物诱导的hiNOS启动子活性。TNF α诱导的hiNOS NF-κ B激活的凝胶位移显示β-连环蛋白突变HA β 18细胞中p50结合减少和NF-κ B报告活性降低。相反，在缺乏β-连环蛋白信号传导的RA β 85细胞中观察到增强的p50结合和增加的NF-κ B报告活性。免疫共沉淀证实β-连环蛋白与p65和p50 NF-κ B蛋白复合。NF-κ B依赖性Traf 1蛋白表达也与β-连环蛋白水平呈负相关。此外，用野生型腺瘤性息肉病稳定转化的SW 480细胞显示出Pcatenin蛋白减少和TNF α诱导的p65 NF-κ B结合以及iNOS和Traf 1增加。表情最后，在肝细胞癌肿瘤样本中，catenin与体内iNOS和Fas表达呈负相关。我们在体外和体内的数据显示，β-连环蛋白信号负相关与马槟榔诱导的hiNOS和其他NF-κ B依赖性基因的表达。这些发现强调了Wnt/β-连环蛋白、NF-κ B和iNOS信号传导在炎症相关癌发生的病理生理学中的复杂作用。[Cancer Res 2009;69（9）：3764-71]
The human inducible nitric oxide synthase (hiNOS) gene is regulated by nuclear factor kappa B (NF-kappa B) and has recently been shown to be a target of the Wnt/beta-catenin pathway. In this study, we tested the hypothesis that Wnt/beta-catenin signaling might regulate cytokine- or tumor necrosis factor a (TNF alpha)-induced hiNOS expression through interaction with NF-kappa B. A cytokine mixture of TNF alpha + interleukin (IL)-1 beta + IFN gamma induced a 2- to 3-fold increase in hiNOS promoter activity in HCT116 and DLD1 colon cells, but produced a 2-fold decrease in SW480 colon cancer cells. A similar differential activity was seen in liver cancer cells (HepG2, Huh7, and Hep3B). Overexpression of beta-catenin produced a dose-dependent decrease in NF-kappa B reporter activity and decreased cytokine mixture-induced hiNOS promoter activity. Gel shift for TNF alpha-induced hiNOS NF-kappa B activation showed decreased p50 binding and decreased NF-kappa B reporter activity in the beta-catenin-mutant HA beta 18 cells. Conversely, enhanced p50 binding and increased NF-kappa B reporter activity were seen in RA beta 85 cells, which lack beta-catenin signaling. Coimmunoprecipitation confirmed that beta-catenin complexed with both p65 and p50 NF-kappa B proteins. NF-kappa B-dependent Traf1 protein expression also inversely correlated with the level of beta-catenin. Furthermore, SW480 cells stably transformed with wild-type adenomatous polyposis coli showed decreased Pcatenin protein and increased TNF alpha-induced p65 NF-kappa B binding as well as iNOS and Traf1. expression. Finally, catenin inversely correlated with iNOS and Fas expression in vivo in hepatocellular carcinoma tumor samples. Our in vitro and in vivo data show that beta-catenin signaling inversely correlates with cytokine-induced hiNOS and other NF-kappa B-dependent gene expression. These findings underscore the complex role of Wnt/beta-catenin, NF-kappa B, and iNOS signaling in the pathophysiology of inflammation-associated carcinogenesis. [Cancer Res 2009;69(9):3764-71]