Organophosphorus chlorpyrifos (CPF) is currently considered an endocrine disruptor (ED), as it can imitate hormone actions both in vitro and in vivo. We recently reported that CPF induces migration and invasion in 2D cultures and changes the expression of key molecular markers involved in epithelial mesenchymal transition in MCF-7 and MDA-MB-231 cell lines. In this study, we investigated whether CPF could behave as a predisposing factor for tumors to become more metastatic and aggressive using 3D culture models. In MCF-7 cells, 0.05 μM CPF induced an increase in the number and size of mammospheres via estrogen receptor alpha (ERα) and c-SRC. Furthermore, 0.05 μM CPF increased the area of spheroids generated from MCF-7 cells, induced invasion using both Matrigel® and type 1 collagen matrices, and increased cell migration capacity via ERα in this 3D model. In turn, 50 μM CPF increased cell migration capacity and invasion using type 1 collagen matrix. In monolayers, CPF increased the phosphorylation and membrane translocation of c-SRC at both concentrations assayed. CPF at 0.05 μM boosted p-AKT, p-GSK-3β and p-P38. While p-AKT rose in a ERα-dependent way, p-GSK-3β was dependent on ERα- and c-SRC, and p-P38 was only dependent on c-SRC. On the other hand, the increase in p-AKT and p-P38 induced by 50 μM CPF was dependent on the c-SRC pathway. We also observed that 0.05 μM CPF increased IGF-1R and IRS-1 expression and that 50 μM CPF induced IGF-1Rβ phosphorylation. In the MDA-MB-231 cell line, 0.05 and 50 μM CPF increased p-c-SRC. Finally, p-AKT and p-GSK-3β were also induced by CPF at 0.05 and 50 μM, and an increase in p-P38 was observed at 50 μM. Taken together, these data provide support for the notion that CPF may represent a risk factor for breast cancer development and progression.

有机磷毒死蜱 (CPF) 目前被认为是一种内分泌干扰物 (ED)，因为它可以在体外和体内模拟激素作用. 我们最近报道了 CPF 在 2D 培养物中诱导迁移和侵袭，并改变 MCF-7 和 MDA-MB-231 细胞系中参与上皮间充质转化的关键分子标志物的表达。在这项研究中，我们使用 3D 培养模型研究了 CPF 是否可以作为肿瘤变得更具转移性和侵袭性的诱发因素。在 MCF-7 细胞中，0.05 μM C​​PF 通过雌激素受体 α (ERα) 和 c-SRC 诱导乳腺球的数量和大小增加。此外，在这个 3D 模型中，0.05 μM C​​PF 增加了 MCF-7 细胞产生的球体的面积，使用 Matrigel® 和 1 型胶原基质诱导侵袭，并通过 ERα 增加了细胞迁移能力。反过来，50 μM C​​PF 使用 1 型胶原基质增加了细胞迁移能力和侵袭能力。在单层中，在两种测定浓度下，CPF 都增加了 c-SRC 的磷酸化和膜易位。0.05 μM 的 CPF 增强了 p-AKT、p-GSK-3β 和 p-P38。虽然 p-AKT 以 ERα 依赖的方式上升，但 p-GSK-3β 依赖于 ERα-和 c-SRC，而 p-P38 仅依赖于 c-SRC。另一方面，50 μM C​​PF 诱导的 p-AKT 和 p-P38 的增加依赖于 c-SRC 途径。我们还观察到 0.05 μM C​​PF 增加 IGF-1R 和 IRS-1 表达，而 50 μM C​​PF 诱导 IGF-1Rβ 磷酸化。在 MDA-MB-231 细胞系中，0.05 和 50 μM C​​PF 增加了 pc-SRC。最后，CPF 在 0.05 和 50 μM 时也诱导了 p-AKT 和 p-GSK-3β，在 50 μM 时观察到 p-P38 的增加。总之，这些数据为CPF可能代表乳腺癌发展和进展的风险因素的观点提供了支持。