Annexin A2 regulates a disintegrin and metalloproteinase 17-mediated ectodomain shedding of pro-tumor necrosis factor-α in monocytes and colon epithelial cells.
Annexin A2 regulates a disintegrin and metalloproteinase 17-mediated ectodomain shedding of pro-tumor necrosis factor-α in monocytes and colon epithelial cells.
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膜联蛋白 A2 调节单核细胞和结肠上皮细胞中解整合素和金属蛋白酶 17 介导的促肿瘤坏死因子-α 的胞外域脱落。
DOI：
10.1097/mib.0b013e318281f43a
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发表时间：
2013
期刊：
影响因子：
--
通讯作者：
Joh T.
中科院分区：
文献类型：
--
作者：
Tsukamoto H;Tanida S;Ozeki K;Ebi M;Mizoshita T;Shimura T;Mori Y;Kataoka H;Kamiya T;Fukuda S;Higashiyama S;Joh T.
背景了解肿瘤坏死因子（tumor necrosis factor，TNF）-α释放的机制是非常重要的，因为TNF-α触发炎症性肠病的发生发展。去整合素和金属蛋白酶（ADAM）17不仅是1型膜锚定蛋白双调蛋白脱落的关键酶，而且是2型蛋白TNF-α脱落的关键酶。然而，ADAM 17切割1型和2型膜锚定蛋白的详细机制尚不清楚。膜联蛋白（ANX）A2参与ADAM17介导的双调蛋白脱落。在这项研究中，我们研究了ANX A2是否参与TNF-α shedding. MethodsWe制备了U937，HT29和HCT 116细胞过表达碱性磷酸酶（AP）标记的proTNF-α和耗尽ADAM 17和ANX A2。我们通过酶联免疫吸附试验和AP试验分别测定白细胞介素-1 β或12-O-十四烷酰佛波醇-13-乙酸酯（TPA）刺激后条件培养基中TNF-α释放浓度和AP活性来评估TNF-α释放和脱落。结果酶联免疫吸附试验和AP试验显示白细胞介素-1 β可诱导HCT 116和HT 29细胞中TNF-α的释放，而TPA可诱导U937细胞中TNF-α的释放。KB-R7785和ADAM17耗竭显著阻断TPA引起的TNF-α释放。ANX A2耗竭显著抑制IL-1 β和TPA引起的TNF-α释放。与此相反，ANX A2耗竭并没有消除ADAM 17介导的双调蛋白和肝素结合表皮生长因子样生长因子脱落。ANX A2与ADAM17直接相关。结论ANX A2与ADAM17密切相关，在TPA释放TNF-α中起重要作用。抑制ANX A2可能是预防炎症性肠病炎症过程中TNF-α释放的一种新的治疗策略。
BackgroundUnderstanding the mechanism of tumor necrosis factor (TNF)-α shedding is important because TNF-α triggers inflammatory bowel disease development. A disintegrin and metalloproteinase (ADAM) 17 is a key enzyme for the shedding of not only the type 1 membrane-anchored protein, amphiregulin, but also the type 2 protein, TNF-α. However, the detailed mechanism by which ADAM17 cleaves type 1 and 2 membrane-anchored proteins is unclear. Annexin (ANX) A2 is involved in ADAM17-mediated amphiregulin shedding. In this study, we examined whether ANX A2 is involved in TNF-α shedding.MethodsWe prepared U937, HT29, and HCT116 cells overexpressing alkaline phosphatase (AP)-tagged proTNF-α and depleted ADAM17 and ANX A2. We assessed TNF-α release and shedding by measuring the TNF-α release concentration and AP activities in conditioned media after interleukin-1β or 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulation by enzyme-linked immunosorbent assay and AP assay, respectively. A direct association of ANX A2 with ADAM17 was examined with immunoprecipitation and Western blotting.ResultsEnzyme-linked immunosorbent assay and AP assay showed interleukin-1β–induced TNF-α shedding in HCT116 and HT29 cells and TPA-induced TNF-α release in U937 cells. KB-R7785 and ADAM17 depletion significantly blocked TNF-α shedding by TPA. ANX A2 depletion significantly inhibited TNF-α shedding by interleukin-1β and TPA. In contrast, ANX A2 depletion did not abrogate ADAM17-mediated amphiregulin and heparin-binding epidermal growth factor-like growth factor shedding. ANX A2 was directly associated with ADAM17.ConclusionsANX A2 was closely associated with ADAM17 and played an important role in TNF-α shedding by TPA. Inhibition of ANX A2 might be a new therapeutic strategy for prevention of TNF-α shedding during inflammatory bowel disease inflammation.