Urriola-Muñoz Paulina, Li Xue, Maretzky Thorsten, McIlwain David R, Mak Tak W, Reyes Juan G, Blobel Carl P, Moreno Ricardo D
Facultad de Ciencias Biológicas, Departamento de Fisiología, Pontificia Universidad Católica de Chile, Santiago, Chile.
Instituto de Química, Pontificia Universidad Católica de Valparaíso, Valparaíso, Chile.
J Cell Physiol. 2018 Mar;233(3):2247-2256. doi: 10.1002/jcp.26097. Epub 2017 Aug 25.
The xenoestrogens bisphenol-A (BPA) and nonylphenol (NP) are endocrine disruptors used in the plastic polymer industry to manufacture different products for human use. Previous studies have suggested a role of these compounds in the shedding of signaling molecules, such as tumor necrosis factor α (TNF-α). The aim of this work was to evaluate the effect of BPA and NP on the sheddase ADAM17 and its newly discovered regulators iRhom1 and iRhom2 in the release of EGFR-ligands. We report that BPA and NP can stimulate the release of the ADAM17-substrates HB-EGF and TGF-α. In cells lacking ADAM17 (Adam17 mEFs) BPA-stimulated release of HB-EGF, but not TGF-α, was strongly reduced, whereas NP-stimulated shedding of HB-EGF and TGF-α was completely abolished. Inactivation of both ADAM17 and the related ADAM10 (Adam10/17 mEFs) completely prevented the release of these substrates. In the absence of iRhom1, BPA- or NP-stimulated release of HB-EGF or TGF-α was comparable to wild-type control mEFs, conversely the BPA-induced release of HB-EGF was abolished in iRhom2 mEFs. The defect in shedding of HB-EGF in iRhom2 mEF cells could be rescued by overexpressing iRhom2. Interestingly, the NP-stimulated release of HB-EGF was not affected by the absence of iRhom2, suggesting that NP could potentially activate both ADAM10 and ADAM17. We tested this hypothesis using betacellulin (BTC), an EGFR-ligand that is a substrate for ADAM10. We found that NP, but not BPA stimulated the release of BTC in Adam17 , iRhom2 , or iRhom1/2 , but not in Adam10/17 cells. Taken together, our results suggest that BPA and NP stimulate the release of EGFR-ligands by differentially activating ADAM17 or ADAM10. The identification of specific effects of these endocrine disruptors on ADAM10 and ADAM17 will help to provide a better understanding of their roles in cell signaling and proinflammatory processes, and provide new potential targets for treatment of reproductive or inflammatory diseases such as asthma or breast cancer that are promoted by xenoestrogens.
异雌激素双酚A（BPA）和壬基酚（NP）是内分泌干扰物，用于塑料聚合物工业中制造各种供人类使用的产品。先前的研究表明这些化合物在信号分子如肿瘤坏死因子α（TNF-α）的脱落中起作用。这项工作的目的是评估BPA和NP对去整合素和金属蛋白酶17（ADAM17）及其新发现的调节因子iRhom1和iRhom2在表皮生长因子受体（EGFR）配体释放中的影响。我们报告称，BPA和NP可刺激ADAM17底物肝素结合表皮生长因子（HB-EGF）和转化生长因子α（TGF-α）的释放。在缺乏ADAM17的细胞（Adam17小鼠胚胎成纤维细胞）中，BPA刺激的HB-EGF释放，但不是TGF-α释放，显著减少，而NP刺激的HB-EGF和TGF-α脱落则完全被消除。ADAM17和相关的ADAM10（Adam10/17小鼠胚胎成纤维细胞）失活完全阻止了这些底物的释放。在缺乏iRhom1的情况下，BPA或NP刺激的HB-EGF或TGF-α释放与野生型对照小鼠胚胎成纤维细胞相当，相反，在iRhom2小鼠胚胎成纤维细胞中，BPA诱导的HB-EGF释放被消除。iRhom2小鼠胚胎成纤维细胞中HB-EGF脱落的缺陷可通过过表达iRhom2来挽救。有趣的是，NP刺激的HB-EGF释放不受iRhom2缺失的影响，这表明NP可能潜在地激活ADAM10和ADAM17。我们使用β细胞素（BTC）（一种EGFR配体，是ADAM10的底物）来验证这一假设。我们发现，NP而非BPA刺激了Adam17、iRhom2或iRhom1/2细胞中BTC的释放，但在Adam10/17细胞中未刺激释放。综上所述，我们的结果表明，BPA和NP通过差异激活ADAM17或ADAM10来刺激EGFR配体的释放。确定这些内分泌干扰物对ADAM10和ADAM17的特定作用将有助于更好地理解它们在细胞信号传导和促炎过程中的作用，并为治疗由异雌激素促进的生殖或炎症性疾病（如哮喘或乳腺癌）提供新的潜在靶点。
