Ren Fei, Ning Hongmei, Ge Yaming, Yin Zhihong, Chen Lingli, Hu Dongfang, Shen Shanshan, Wang Xinrui, Wang Siting, Li Rongbo, He Junping
College of Veterinary Medicine, Shanxi Agricultural University, Jinzhong, Shanxi 030801, China; College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, Henan 453003, China.
College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, Henan 453003, China.
Toxicology. 2022 Mar 30;470:153138. doi: 10.1016/j.tox.2022.153138. Epub 2022 Feb 24.
Bisphenol A (2,2-bis(4'-hydroxyphenyl) propane, BPA) is a well-known endocrine-disrupting compound that is widely used in various daily products and exhibits embryonic development toxicity and genotoxicity. However, the affected signaling pathways involved in embryonic development especially the interactions of involved proteins remain unclear. In our previous study (Ge et al., 2021), BPA induces DNA damage and apoptosis in Xenopus embryos, resulting in multiple malformations of larvae. However, the signaling pathways induced for apoptosis response to DNA damage are still not well elucidated. Here, we systematically elucidated the enriched pathways affected by BPA and illustrated the interactions of involved proteins. Results indicated that BPA affected multiple embryonic development pathways including Hippo, TGF-β, Wnt, and Notch pathways. Furthermore, the protein-protein interaction network suggested that the c-Abl/YAP/p73 pathway may play a key role in apoptosis induction in response to DNA damage. P19 embryonal carcinoma stem cells, as a developmental toxicity model, were treated with different BPA concentrations to establish an in vitro model to verify the role of the c-Abl/YAP/p73 pathway in apoptosis. BPA triggered DNA damage and significantly upregulated the expression levels of c-Abl, phosphorylated YAP, phosphorylated p73, and cleaved caspase-3 protein (p < 0.05), thus decreasing cell viability and transcriptionally activating the p73 target genes Bax and Puma. These data suggested that BPA activated the c-Abl/YAP/p73 pathway in response to DNA damage. Imatinib, an inhibitor of tyrosine kinase c-Abl, significantly downregulated the elevated expression levels of p-YAP, p-p73 and cleaved caspase-3 (p < 0.05) caused by BPA and then ameliorated the cell index of P19 cells in the BPA-treated group. Therefore, this substance restrained the phosphokinase activity of c-Abl and suppressed the c-Abl/YAP/p73 pathway. Results showed that the c-Abl/YAP/p73 pathway served as a mechanism for caspase-3 activation that induced the apoptosis response to DNA damage stress.
双酚A（2,2-双（4'-羟基苯基）丙烷，BPA）是一种著名的内分泌干扰化合物，广泛应用于各种日常用品中，并具有胚胎发育毒性和基因毒性。然而，参与胚胎发育的受影响信号通路，尤其是相关蛋白质的相互作用仍不清楚。在我们之前的研究（Ge等人，2021年）中，BPA诱导非洲爪蟾胚胎中的DNA损伤和凋亡，导致幼虫出现多种畸形。然而，针对DNA损伤的凋亡反应所诱导的信号通路仍未得到很好的阐明。在此，我们系统地阐明了受BPA影响的富集通路，并说明了相关蛋白质的相互作用。结果表明BPA影响多个胚胎发育通路，包括Hippo、TGF-β、Wnt和Notch通路。此外，蛋白质-蛋白质相互作用网络表明，c-Abl/YAP/p73通路可能在响应DNA损伤的凋亡诱导中起关键作用。作为发育毒性模型的P19胚胎癌细胞系用不同浓度的BPA处理，以建立体外模型来验证c-Abl/YAP/p73通路在凋亡中的作用。BPA引发DNA损伤，并显著上调c-Abl、磷酸化YAP、磷酸化p73和裂解型半胱天冬酶-3蛋白的表达水平（p < 0.05），从而降低细胞活力并转录激活p73靶基因Bax和Puma。这些数据表明BPA在响应DNA损伤时激活了c-Abl/YAP/p73通路。酪氨酸激酶c-Abl的抑制剂伊马替尼显著下调了由BPA引起的p-YAP、p-p73和裂解型半胱天冬酶-3升高的表达水平（p < 0.05），然后改善了BPA处理组中P19细胞的细胞指数。因此，该物质抑制了c-Abl的磷酸激酶活性并抑制了c-Abl/YAP/p73通路。结果表明，c-Abl/YAP/p73通路作为半胱天冬酶-3激活的一种机制，诱导了对DNA损伤应激的凋亡反应。
