多维方法评价全氟辛酸对人结直肠癌细胞的毒性效应  

Evaluation of the toxicity of perfluorooctanoic acid toward human colorectal cancer cells using multi-dimensional approaches

作　　者：张睿佳 林颖诗 涂蓝尹 陈梓潼 张维蔚 栾天罡[2] 陈保卫 ZHANG Ruijia;LIN Yingshi;TU Lanyin;CHEN Zitong;ZHANG Weiwei;LUAN Tiangang;CHEN Baowei(School of Life and Health Sciences,Hainan University,Haikou 570228,China;School of Life Sciences,Sun Yat-sen University,Guangzhou 510275,China;School of Marine Sciences,Sun Yat-sen University,Zhuhai 519082,China;School of Artificial Intelligence,Sun Yat-sen University,Zhuhai 519082,China;Guangzhou Center for Disease Control and Prevention,Guangzhou 510440,China)

机构地区：[1]海南大学生命健康学院,海南海口570228 [2]中山大学生命科学学院,广东广州510275 [3]中山大学海洋科学学院,广东珠海519082 [4]中山大学人工智能学院,广东珠海519082 [5]广州市疾病控制中心,广东广州510440

出　　处：《色谱》2025年第1期96-103,共8页Chinese Journal of Chromatography

基　　金：国家自然科学基金(22206207,22127810,22276224);广州市科技计划(2024A04J3793,2024A03J0414)。

摘　　要：全氟辛酸(PFOA)的暴露与溃疡性结肠炎的发生密切相关,但目前缺少PFOA暴露对人结直肠癌细胞(HCT116)产生毒性效应的相关分子机理研究。本研究从细胞毒性表型、细胞呼吸和代谢相关基因转录水平3个层次评价了PFOA对HCT116的毒性效应。首先,利用水溶性甲臜化合物(MTS)来评价PFOA暴露对HCT116细胞相对活性的影响;随后,利用细胞外流量分析仪对HCT116的线粒体呼吸活性进行测定;最后,用定量即时聚合酶链式反应(qPCR)对HCT116中代谢相关基因的转录水平进行检测。细胞毒性实验结果表明,与对照组相比,经高浓度(300μmol/L)PFOA暴露48 h后,HCT116的细胞活性受到显著抑制(p<0.01),并在G0/G1细胞周期受到阻滞,而低浓度(30、50μmol/L)的PFOA反而提高了细胞相对活性;低浓度(50μmol/L)的PFOA能够促进HCT116的线粒体呼吸活性。利用自主开发的Metabolic Gene and Pathway Query检索软件和比较毒理基因组学数据库,本研究发现代谢相关基因二肽酶1(DPEP1)和鞘甘氨酸-1激酶(SPHK1)与PFOA引起的溃疡性结肠炎相关。qPCR实验结果表明,高浓度(300μmol/L)PFOA能够显著诱导DPEP1和SPHK1的转录表达上调(约8~10倍),低浓度(50μmol/L)PFOA未引起DPEP1和SPHK1转录表达水平的变化。本文发现细胞线粒体呼吸活性是评价低浓度PFOA干扰效应的一个敏感指标,DPEP1和SPHK1介导的细胞代谢过程可能是PFOA引起肠道毒性的潜在机制。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 300μmol/L PFOA for 48 h(p<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 50μmol/L.Two genes related to cellular metabolism(dipeptidase 1(DPEP 1)and sphingosine kinase 1(SPHK 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 DPEP 1 and SPHK 1 expression levels were enhanced by 8-10 times in HCT116 exposed to 300μmol/L PFOA relative to the control,whereas this trend was not observed for HCT116 exposed to 50μmol/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 DPEP 1 and SPHK 1 may be involved in the development of PFOA-induced ulcerative colitis.Future studies should investigate the relationships between changes in metabolism-related genes(DPEP 1 and SPHK 1)and the mitochondrial respiratory activities of intestinal cells,and v

关 键 词：全氟辛酸 人结直肠肠道毒性 代谢紊乱 代谢基因和通路检索软件 

分 类 号：O658[理学—分析化学]