6PPD and 6PPD Quinone Induce Endometrial Cell Dysfunction via Activating ERα and GPER at Human-Relevant Levels

6PPD and 6PPD Quinone Induce Endometrial Cell Dysfunction via Activating ERα and GPER at Human-Relevant LevelsClick to copy article linkArticle link copied! - Lei TangLei TangCollege of Environment and Ecology, Hunan Agricultural University, 1 Nongda Road, Furong District, Changsha 410128, ChinaMore by Lei Tang - Xin LiXin LiCollege of Environment and Ecology, Hunan Agricultural University, 1 Nongda Road, Furong District, Changsha 410128, ChinaMore by Xin Li - Shi-Ye ZhuShi-Ye ZhuCollege of Environment and Ecology, Hunan Agricultural University, 1 Nongda Road, Furong District, Changsha 410128, ChinaMore by Shi-Ye Zhu - Yu-Ke WanYu-Ke WanCollege of Environment and Ecology, Hunan Agricultural University, 1 Nongda Road, Furong District, Changsha 410128, ChinaMore by Yu-Ke Wan - Shi-Ke ZhangShi-Ke ZhangCollege of Environment and Ecology, Hunan Agricultural University, 1 Nongda Road, Furong District, Changsha 410128, ChinaMore by Shi-Ke Zhang - Jie-Zhi MaJie-Zhi MaDepartment of Obstetrics and Gynecology, Xiangya Third Hospital, Central South University, Changsha 410013, ChinaMore by Jie-Zhi Ma - Yuan YangYuan YangCollege of Environment and Ecology, Hunan Agricultural University, 1 Nongda Road, Furong District, Changsha 410128, ChinaMore by Yuan Yang - An-Wei ChenAn-Wei ChenCollege of Environment and Ecology, Hunan Agricultural University, 1 Nongda Road, Furong District, Changsha 410128, ChinaMore by An-Wei Chen - Lin Luo*Lin Luo*Email: [email protected]College of Environment and Ecology, Hunan Agricultural University, 1 Nongda Road, Furong District, Changsha 410128, ChinaMore by Lin Luo - Lin-Ying Cao*Lin-Ying Cao*Email: [email protected]. Tel./Fax: 86 0731-84618166.College of Environment and Ecology, Hunan Agricultural University, 1 Nongda Road, Furong District, Changsha 410128, ChinaMore by Lin-Ying Cao Abstract The widespread environmental prevalence of tire-derived N-(1,3-dimethylbutyl)-N′-phenyl-p-phenylenediamine (6PPD) and 6PPD-quinone (6PPD-Q) has provoked public concern about their health risks. This study aimed to investigate the potential of 6PPD and 6PPD-Q to induce endometrial cell dysfunction through nuclear estrogen receptor (ER) and G-protein-coupled estrogen receptor (GPER) signaling pathways. Fluorescence competitive binding and reporter gene assays revealed that 6PPD and 6PPD-Q selectively bound to ERα (not ERβ) and activated ER transcriptional activity, with the lowest observed effective concentrations (LOECs) of 500 and 10 nM, respectively. Calcium mobilization assays further demonstrated that both 6PPD and 6PPD-Q activated the GPER nongenomic pathway in a concentration-dependent manner (LOEC = 1 nM). 6PPD-Q exhibited stronger ERα and GPER activation potency than 6PPD, which was explained well by molecular dynamics simulation. 6PPD and 6PPD-Q stimulated endometrial cell proliferation via ERα/GPER signaling pathways, mechanistically linked to Cyclin D1/Ki67 upregulation. Furthermore, 6PPD/6PPD-Q promoted endometrial cell migration through an ERα/GPER-regulated epithelial-mesenchymal transition and inflammatory responses. Notably, the LOECs for these functional disruptions reached nanomolar levels relevant to human exposure. Collectively, we elucidated the molecular initial events and downstream key events of 6PPD/6PPD-Q-induced endometrial cell dysfunction, which implied their threat to the reproductive system and provided novel perspectives for their health risk evaluation. 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