Evaluation of the toxicity of perfluorooctanoic acid toward human colorectal cancer cells using multi-dimensional approaches
While human exposure to perfluorooctanoic acid(PFOA)can lead to ulcerative coli-tis,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(p＜0.01),with most cells re-tained 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(DPEP1)and sphingosine kinase 1(SPHK1))were found to be re-lated 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 DPEP1 and SPHK1 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 ob-served for HCT116 exposed to 50 μmol/L PFOA.Collectively,these results suggest that the re-spiratory activity of cellular mitochondria may serve as an index for determining the interfer-ence effects associated with PFOA and that metabolic pathways mediated by DPEP1 and SPHK1 may be involved in the development of PFOA-induced ulcerative colitis.Future studies should investigate the relationships between changes in metabolism-related genes(DPEP1 and SPHK1)and the mitochondrial respiratory activities of intestinal cells,and verify the roles played by the DPEP1 and SPHK1 genes in PFOA-induced intestinal inflammation using in-vivo models.
perfluorooctanoic acid(PFOA)human colorectal enterotoxicitymetabolic disor-derMetabolic Gene and Pathway Query software
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