Zhang Rui-Jia, Lin Ying-Shi, Tu Lan-Yin, Chen Zi-Tong, Zhang Wei-Wei, Luan Tian-Gang, Chen Bao-Wei
School of Life and Health Sciences, Hainan University, Haikou 570228, China.
School of Life Sciences, Sun Yat-sen University, Guangzhou 510275, China.
Se Pu. 2025 Jan;43(1):96-103. doi: 10.3724/SP.J.1123.2024.05022.
While human exposure to perfluorooctanoic acid (PFOA) can lead to ulcerative colitis, the molecular mechanisms responsible for PFOA-induced intestinal toxicity are unclear. Herein, we examined the toxicity of PFOA toward human colorectal cancer cells (HCT116) from three dimensions: the cytotoxic phenotype, cell respiration, and transcription levels of metabolism-related genes. Formazan was used to assess how PFOA exposure affects HCT116-cell relative viability, after which the mitochondrial respiratory activities of these cells were determined by analyzing extracellular flux. The quantitative real-time polymerase chain reaction (qPCR) method was used to detect metabolism-related gene expression levels. The cytotoxicity assay revealed that the HCT116 showed significantly inhibited relative activities compared to those of the control when exposed to 300 μmol/L PFOA for 48 h (<0.01), with most cells retained at the G0/G1 stage. In contrast, the mitochondrial respiratory activities of the HCT116 were promoted by concentrations of PFOA as low as 50 μmol/L. Two genes related to cellular metabolism (dipeptidase 1 () and sphingosine kinase 1 ()) were found to be related to the PFOA-promoted formation of ulcerative colitis using our self-developed Metabolic Gene and Pathway Query software and Comparative Toxicogenomics Database (CTD). The qPCR studies revealed that and expression levels were enhanced by 8-10 times in HCT116 exposed to 300 μmol/L PFOA relative to the control, whereas this trend was not observed for HCT116 exposed to 50 μmol/L PFOA. Collectively, these results suggest that the respiratory activity of cellular mitochondria may serve as an index for determining the interference effects associated with PFOA and that metabolic pathways mediated by and may be involved in the development of PFOA-induced ulcerative colitis. Future studies should investigate the relationships between changes in metabolism-related genes ( and ) and the mitochondrial respiratory activities of intestinal cells, and verify the roles played by the and genes in PFOA-induced intestinal inflammation using in-vivo models.
虽然人类接触全氟辛酸（PFOA）会导致溃疡性结肠炎，但PFOA诱导肠道毒性的分子机制尚不清楚。在此，我们从细胞毒性表型、细胞呼吸和代谢相关基因的转录水平三个维度研究了PFOA对人结肠癌细胞（HCT116）的毒性。用甲臜评估PFOA暴露如何影响HCT116细胞的相对活力，之后通过分析细胞外通量来测定这些细胞的线粒体呼吸活性。采用定量实时聚合酶链反应（qPCR）方法检测代谢相关基因的表达水平。细胞毒性试验表明，当暴露于300 μmol/L PFOA 48小时时，HCT116的相对活性与对照组相比显著受到抑制（<0.01），大多数细胞停滞在G0/G1期。相反，低至50 μmol/L的PFOA浓度就能促进HCT116的线粒体呼吸活性。利用我们自行开发的代谢基因和通路查询软件以及比较毒理基因组学数据库（CTD），发现两个与细胞代谢相关的基因（二肽酶1（）和鞘氨醇激酶1（））与PFOA促进溃疡性结肠炎的形成有关。qPCR研究表明，相对于对照组，暴露于300 μmol/L PFOA的HCT116中 和 的表达水平提高了8至10倍，而暴露于50 μmol/L PFOA的HCT116中未观察到这种趋势。总体而言，这些结果表明，细胞线粒体的呼吸活性可能作为确定与PFOA相关的干扰效应的指标，并且由 和 介导的代谢途径可能参与PFOA诱导的溃疡性结肠炎的发展。未来的研究应调查代谢相关基因（ 和 ）的变化与肠道细胞线粒体呼吸活性之间的关系，并使用体内模型验证 和 基因在PFOA诱导的肠道炎症中所起的作用。
