Abstract
This study aimed to explore the effects of perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) on apoptosis and cell cycle in a zebrafish (Danio rerio) liver cell line (ZFL). Treatment groups included a control group, PFOA-IC50, PFOA-IC80, PFOS-IC50 and PFOS-IC80 groups. IC50 and IC80 concentrations were identified by cellular modeling and MTT assays. mRNA levels of p53, Bcl-2, Bax, Caspase-3 and NF-κB p65 were detected by qPCR. Cell apoptosis and cell cycle were detected by flow cytometry and the protein levels of p53, Bcl-2, Bax, Caspase-3 and NF-κB p65 were determined by western blotting. Both PFOA and PFOS inhibited the growth of zebrafish liver cells, and the inhibition rate of PFOS was higher than that of PFOA. Bcl-2 expression levels in the four groups were significantly higher than the control group and Bcl-2 increased significantly in the PFOA-IC80 group. However, the expression levels of Bax in the four treatment groups were higher than the control group. The percentage of cell apoptosis increased significantly with the treatment of PFOA and PFOS (p < 0.05). Cell cycle and cell proliferation were blocked in both the PFOA-IC80 and PFOS-IC80 groups, indicating that PFOA-IC80 and PFOS-IC50 enhanced apoptosis in ZFL cells.
Highlights
Multicellular animals dispose of unneeded or damaged cells in an ordered fashion by apoptosis [1]
MTT assay different concentrations of perfluorooctanoic acid (PFOA) or perfluorooctane sulfonate (PFOS), and the inhibition rate was determined by an MTT assay
P65 expression level was induced in all groups except the NF-κB p65 expression level was induced in all groups except the PFOA-IC8080 group. These results showed that PFOA and PFOS treatment could affect the expression of apoptosis related genes, showed that PFOA and PFOS treatment could affect the expression of apoptosis related genes, including p53, Bcl-2, Bax, Caspase-3 and NF-κB p65
Summary
Multicellular animals dispose of unneeded or damaged cells in an ordered fashion by apoptosis [1]. Apoptosis is a programmed cell death pathway that removes damaged cells to protect the body, and it is especially important in vertebrate embryonic development [2]. The relationship between stress-induction and apoptosis during the development has been confirmed by experimental studies using zebrafish (Danio rerio) embryos, and researches on apoptotic genes, including the Bcl-2 gene family, p53, and the caspase family, are still under the spotlight [3,4,5,6]. Zebrafish is considered to be an effective animal model to reveal the relationship between stress-induction and apoptosis. Perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS) are two of the most widely used PFCs. PFOA and PFOS have already been considered as emerging persistent
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