Department of Internal Medicine and Therapeutics, University of Pavia, 27100, Italy.
Department of Internal Medicine and Therapeutics, University of Pavia, 27100, Italy; Istituti Clinici Scientifici Maugeri IRCCS, Unit of Internal Medicine and Endocrinology, Laboratory for Endocrine Disruptors, 27100 Pavia, Italy.
Environ Int. 2024 May;187:108717. doi: 10.1016/j.envint.2024.108717. Epub 2024 May 6.
Exposure to environmental pollutants is suspected to be one of the potential causes accounting for the increase in thyroid cancer (TC) incidence worldwide. Among the ubiquitous pollutants, per-polyfluoroalkyl substances (PFASs), were demonstrated to exert thyroid disrupting effects. Perfluoroalkyl carboxylates (PFCAs) represent a subgroup of PFAS and include perfluoro carboxylic acids (PFOA and PFHxA) and perfluoropolyether carboxylic acid (C6O4). The potential relationship between exposure to PFCAs and TC was not yet fully elucidated. This in vitro study investigated whether certain PFCAs (C6O4, PFOA, and PFHxA) can influence the composition of TC microenvironment.
Two models of normal thyroid cells in primary cultures: Adherent (A-NHT) and Spheroids (S-NHT) were employed. A-NHT and S-NHT were exposed to C6O4, PFOA or PFHxA (0; 0.01; 0.1, 1; 10; 100; 1000 ng/mL) to assess viability (WST-1 and AV/PI assay), evaluate spherification index (SI) and volume specifically in S-NHT. CXCL8 and CCL2 (mRNA and protein), and EMT-related genes were assessed in both models after exposure to PFCAs.
PFHxA reduced the viability of both A-NHT and S-NHT. None of the PFCAs interfered with the volume or spherification process in S-NHT. CXCL8 and CCL2 mRNA and protein levels were differently up-regulated by each PFCAs, being PFOA and PFHxA the stronger inducers. Moreover, among the tested PFCAs, PFHxA induced a more consistent increase in the mRNA levels of EMT-related genes.
This is the first evaluation of the effects of exposure to PFCAs on factors potentially involved in establishing the TC microenvironment. PFHxA modulated the TC microenvironment at three levels: cell viability, pro-tumorigenic chemokines, and EMT-genes. The results provide further evidence of the pro-tumorigenic effect of PFOA. On the other hand, a marginal effect was observed for C6O4 on pro-tumorigenic chemokines.
环境污染暴露被怀疑是导致全球甲状腺癌 (TC) 发病率上升的潜在原因之一。在无处不在的污染物中，全氟烷基物质 (PFAS) 被证明具有甲状腺干扰作用。全氟烷基羧酸 (PFCAs) 是 PFAS 的一个亚组，包括全氟己酸 (PFOA 和 PFHxA) 和全氟聚醚羧酸 (C6O4)。接触 PFCAs 与 TC 之间的潜在关系尚未完全阐明。这项体外研究调查了某些 PFCAs (C6O4、PFOA 和 PFHxA) 是否会影响 TC 微环境的组成。
使用两种原代培养的正常甲状腺细胞模型：贴壁 (A-NHT) 和球体 (S-NHT)。将 A-NHT 和 S-NHT 暴露于 C6O4、PFOA 或 PFHxA(0；0.01；0.1、1；10；100；1000 ng/mL) 中，以评估活力 (WST-1 和 AV/PI 测定)，评估 S-NHT 中的球体化指数 (SI) 和体积。在接触 PFCAs 后，评估两种模型中的 CXCL8 和 CCL2(mRNA 和蛋白质)和 EMT 相关基因。
PFHxA 降低了 A-NHT 和 S-NHT 的活力。没有一种 PFCAs 会干扰 S-NHT 的体积或球体化过程。每种 PFCAs 都不同程度地上调了 CXCL8 和 CCL2 的 mRNA 和蛋白质水平，其中 PFOA 和 PFHxA 的诱导作用更强。此外，在测试的 PFCAs 中，PFHxA 诱导 EMT 相关基因的 mRNA 水平更一致地增加。
这是首次评估接触 PFCAs 对潜在参与建立 TC 微环境的因素的影响。PFHxA 在三个层面上调节 TC 微环境：细胞活力、促肿瘤趋化因子和 EMT-基因。结果进一步证明了 PFOA 的促肿瘤作用。另一方面，C6O4 对促肿瘤趋化因子的作用很小。
