S100B Protein Stimulates Proliferation and Angiogenic Mediators Release through RAGE/pAkt/mTOR Pathway in Human Colon Adenocarcinoma Caco-2 Cells
S100B Protein Stimulates Proliferation and Angiogenic Mediators Release through RAGE/pAkt/mTOR Pathway in Human Colon Adenocarcinoma Caco-2 Cells
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DOI：
10.3390/ijms20133240
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发表时间：
2019-07-01
影响因子：
5.6
通讯作者：
Esposito, Giuseppe
中科院分区：
文献类型：
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作者：
Seguella, Luisa;Capuano, Riccardo;Esposito, Giuseppe
慢性炎症和血管生成与结肠癌发生有关。肠胶质细胞来源的 S100B 蛋白被认为是连接结肠炎症和癌症的理想桥梁，因为它具有通过晚期糖基化终末产物 (RAGE) 信号传导受体上调核因子 -kappaB (NF-kappa B) 转录和隔离野生型促凋亡野生型 (wt)p53 的双重能力。然而，其对癌细胞的促血管生成作用仍未得到研究。为此，我们分别通过以下方法评估了单独的外源 S100B (0.05-5 mu M) 蛋白或在 S100B 阻断单克隆抗体 (mAb)（1:10(5)-1:10(4) v/v 稀释）存在下对 (1) 培养的 Caco-2 细胞增殖、迁移和侵袭性的影响。 3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四唑溴化物 (MTT)-甲臜、伤口愈合和基质胶侵袭测定以及 (2) 通过 ELISA 和免疫荧光分析其对促血管生成因子（如血管内皮生长因子 (VEGF)）释放的影响。然后通过免疫印迹分析研究单独的 S100B 或在 S100BmAb 存在下对 RAGE/pAkt/哺乳动物雷帕霉素靶点 (mTOR) 信号通路的影响。我们的结果表明，S100B 通过释放促血管生成 VEGF 和 NO，同时显着降低 RAGE-p38 丝裂原激活蛋白激酶 (MAPK)/pAkt-mTOR 和缺氧诱导因子 1-α (HIF1 α) 途径介导的 wtp53 表达，从而显着增加 Caco-2 细胞的增殖和侵袭力。这种效应被 S100BmAb 抵消，表明 S100B 靶向是抑制结肠癌增殖和血管生成的潜在方法。
Chronic inflammation and angiogenesis are associated with colonic carcinogenesis. Enteric glia-derived S100B protein has been proposed as an ideal bridge, linking colonic inflammation and cancer, given its dual ability to up-regulate nuclear factor-kappaB (NF-kappa B) transcription via receptor for advanced glycation end products (RAGE) signaling and to sequestrate wild type pro-apoptotic wild type (wt)p53. However, its pro-angiogenic effects on cancer cells are still uninvestigated. To this aim, we evaluated the effect of exogenous S100B (0.05-5 mu M) protein alone or in the presence of S100B blocking monoclonal antibody (mAb) (1:10(5)-1:10(4) v/v diluted) on (1) cultured Caco-2 cells proliferation, migration and invasiveness in vitro, respectively by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT)-formazan, wound healing and matrigel invasion assays and (2) its effect on the release of pro-angiogenic factors, such as vascular endothelial growth factor (VEGF) by ELISA and immunofluorescence analyses. The effect of S100B alone or in the presence of S100BmAb was then investigated on RAGE/pAkt/mammalian target of rapamycin (mTOR) signaling pathway by immunoblot analysis. Our results showed that S100B markedly increases proliferation and invasiveness of Caco-2 cells, through the release of pro-angiogenic VEGF and NO paralleled to a significant decrease of wtp53 expression mediated by RAGE-p38 mitogen-activated protein kinase (MAPK)/pAkt-mTOR and hypoxia-inducible factor 1-alpha (HIF1 alpha) pathways. Such effects were counteracted by S100BmAb, indicating that S100B targeting is a potential approach to inhibit colon carcinoma proliferation and angiogenesis.