Department of Internal Medicine and Therapeutics, University of Pavia, Pavia (PV), 27100, Italy.
Department of Clinical and Experimental Medicine, Endocrinology Unit, University of Pisa (PI), via Paradisa 2, 56124, Pisa, Italy.
Environ Sci Pollut Res Int. 2023 May;30(23):63522-63534. doi: 10.1007/s11356-023-26797-6. Epub 2023 Apr 13.
Industrial chemical PFAS are persistent pollutants. Long chain PFAS were taken out of production due to their risk for human health, however, new congeners PFAS have been introduced. The in vitro effects of the long-chain PFOA, the short-chain PFHxA and the new-generation C6O4 were evaluated in normal and in thyroid cancer cell lines in terms of cell viability and proliferation, and secretion of a pro-tumorigenic chemokine (CXCL8), both at the mRNA and at the protein level. The Nthy-ory 3-1 normal-thyroid cell line, the TPC-1 and the 8505C (RET/PTC rearranged and BRAFV600e mutated, respectively) thyroid-cancer cell lines were exposed to increasing concentrations of each PFAS in a time-course. We evaluated viability using WST-1 (confirmed by AnnexinV/PI) and proliferation using the cristal-violet test. To evaluate CXCL8 mRNA we used RT-PCR and measured CXCL8 in the supernatants by ELISA. The exposure to none PFAS did not affect thyroid cells viability (except for a reduction of 8505C cells viability after 144 h) or proliferation. Individual PFAS differently modulated CXCL8 mRNA and protein level. PFOA increased CXCL8 both at mRNA and protein level in the three cell lines; PFHxA increased CXCL8 mRNA in the three cell lines, but increased the protein only in TPC-1 cells; C6O4 increased the CXCL8 mRNA only in thyroid cancer cell lines, but never increased the CXCL8 protein. The results of the present study indicate that the in vitro exposure to different PFAS may modulate both at the mRNA and secreted protein levels of CXCL8 in normal and cancer thyroid cells. Strikingly different effects emerged according to the specific cell type and to the targeted analyte (CXCL8 mRNA or protein).
工业用化学物质全氟辛酸（PFAS）是持久性污染物。由于对人类健康存在风险，长链 PFAS 已被淘汰，但新的同类物 PFAS 已被引入。本研究旨在评估长链 PFOA、短链 PFHxA 和新一代 C6O4 对正常甲状腺细胞系和甲状腺癌细胞系（Nthy-ory 3-1、TPC-1 和 8505C，分别为 RET/PTC 重排和 BRAFV600e 突变）的体外细胞活力和增殖以及促肿瘤生成趋化因子（CXCL8）的分泌的影响，评估指标包括 mRNA 和蛋白水平。在时间进程中，用不同浓度的每种 PFAS 处理 Nthy-ory 3-1 正常甲状腺细胞系、TPC-1 和 8505C（RET/PTC 重排和 BRAFV600e 突变）甲状腺癌细胞系。用 WST-1（通过 AnnexinV/PI 确认）评估细胞活力，用结晶紫试验评估增殖。通过 RT-PCR 评估 CXCL8 mRNA，通过 ELISA 测量上清液中的 CXCL8。没有一种 PFAS 会影响甲状腺细胞活力（除了 8505C 细胞活力在 144 小时后降低）或增殖。单独的 PFAS 以不同的方式调节 CXCL8 mRNA 和蛋白水平。PFOA 在三种细胞系中均增加 CXCL8 的 mRNA 和蛋白水平；PFHxA 在三种细胞系中增加 CXCL8 的 mRNA，但仅在 TPC-1 细胞中增加 CXCL8 的蛋白；C6O4 仅在甲状腺癌细胞系中增加 CXCL8 的 mRNA，但从未增加 CXCL8 的蛋白。本研究结果表明，不同 PFAS 的体外暴露可能会在正常和甲状腺癌细胞中同时调节 CXCL8 的 mRNA 和分泌蛋白水平。根据特定的细胞类型和靶向分析物（CXCL8 mRNA 或蛋白），出现了明显不同的效果。
