Abstract：
Triclocarban (TCC) and triclosan (TCS) have been detected ubiquitously in human body and evoked increasing concerns. This study aimed to reveal the induction risks of TCC and TCS on triple negative breast cancer through non-genomic GPER-mediated signaling pathways. Molecular simulation indicated that TCC exhibited higher GPER binding affinity than TCS theoretically. Calcium mobilization assay displayed that TCC/TCS activated GPER signaling pathway with the lowest observed effective concentrations (LOEC) of 10 nM/100 nM. TCC and TCS also upregulated MMP-2/9, EGFR, MAPK3 but downregulated MAPK8 via GPER-mediated signaling pathway. Proliferation assay showed that TCC/TCS induced 4 T1 breast cancer cells proliferation with the LOEC of 100 nM/1000 nM. Wound-healing and transwell assays showed that TCC/TCS promoted 4 T1 cells migration in a concentration-dependent manner with the LOEC of 10 nM. The effects of TCC on breast cancer cells proliferation and migration were stronger than TCS and both were regulated by GPER. TCC/TCS induced migratory effects were more significantly than proliferative effect. Mechanism study showed that TCC/TCS downregulated the expression of epithelial marker (E-cadherin) but upregulated mesenchymal markers (snail and N-cadherin), which was reversed by GPER inhibitor G15. These biomarkers results indicated that TCC/TCS-induced 4 T1 cells migration was a classic epithelial to mesenchymal transition mechanism regulated by GPER signaling pathway. Orthotopic tumor model verified that TCC promoted breast cancer in-situ tumor growth and distal tissue metastasis via GPER-mediated signaling pathway at human-exposure level of 10 mg/kg/d. TCC-induced tissue metastasis of breast cancer was more significantly than in-situ tumor growth. Overall, we demonstrated for the first time that TCC/TCS could activate the GPER signaling pathways to induce breast cancer progression.
摘要：
三氯卡班（TCC）和三氯生（TCS）已在人体中普遍存在，并引起越来越多的关注。本研究旨在揭示TCC和TCS通过非基因组GPER介导的信号通路对三阴性乳腺癌的诱导风险。分子模拟表明，TCC在理论上表现出比TCS更高的GPER结合亲和力。钙动员测定显示TCC/TCS激活GPER信号传导途径,观察到的最低有效浓度(LOEC)为10nM/100nM。TCC和TCS也上调MMP-2/9，EGFR，MAPK3但通过GPER介导的信号通路下调MAPK8。增殖试验显示TCC/TCS诱导4T1乳腺癌细胞增殖,LOEC为100nM/1000nM。伤口愈合和transwell测定显示TCC/TCS以浓度依赖性方式促进4T1细胞迁移,LOEC为10nM。TCC对乳腺癌细胞增殖和迁移的影响强于TCS，且均受GPER调控。TCC/TCS诱导的迁移效应比增殖效应更显著。机制研究表明，TCC/TCS下调上皮标志物（E-cadherin）的表达，但上调间充质标志物（蜗牛和N-cadherin）的表达，被GPER抑制剂G15逆转。这些生物标志物结果表明，TCC/TCS诱导的4T1细胞迁移是GPER信号通路调控的一种经典的上皮间质转化机制。原位肿瘤模型证实，在人暴露水平为10mg/kg/d时，TCC通过GPER介导的信号通路促进乳腺癌原位肿瘤生长和远端组织转移。TCC诱导的乳腺癌组织转移比原位肿瘤生长更显著。总的来说,我们首次证明TCC/TCS可以激活GPER信号通路诱导乳腺癌进展.
