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ACTIVATION ASSAY FOR PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR- ALPHA (PPARÁ) BY PERFLUOROALKYL ACIDS (PFAAS) IN COS-1 CELLS
Citation:
WOLF, C. J., M. TAKACS, AND B. D. ABBOTT. ACTIVATION ASSAY FOR PEROXISOME PROLIFERATOR-ACTIVATED RECEPTOR- ALPHA (PPARÁ) BY PERFLUOROALKYL ACIDS (PFAAS) IN COS-1 CELLS. Presented at Current Concepts in Toxicology Workshop, Arlington, VA, February 14 - 16, 2007.
Description:
PFAAs have been found to elicit various physiological effects including peroxisome proliferation, indicating the mechanism of action for these chemicals could involve PPAR. This study investigates the ability of PFAAs to bind and activate mouse and human PPARα in COS-1 cells. COS-1 cells were transiently transfected with a single plasmid that contains two contstructs; one containing the ligand binding domain of the mouse or human PPARα linked to a GAL-4 DNA binding domain, and the other a PPARα-responsive luciferase promoter-reporter construct. Transfected cells were exposed to water or DMSO as negative vehicle controls, PPAR-α agonist WY14643 as a positive control, or to PFAAs in increasing doses, for 24 hours. Activation of the plasmid was measured by chemiluminescence on a luminometer. Relative luminometer units were log 10 transformed and normalized to the mean of the negative control values. Differences in activation were evaluated using analysis of variance and Bonferroni or Dunnett's multiple comparison tests. In all cases, WY14683 significantly increased activation of PPAR¿ plasmid. Perfluorooctanoic acid (PFOA) increased activation of human and mouse PPARα plasmid in a dose-dependent fashion, and the lowest significant increase was 10 µM. Perfluorodecanoate (PFDA) induced activation of murine PPARα plasmid in a dose-dependent fashion with low standard errors, and the lowest significant increase was 10 µM, while PFDA did not induce significant activation of human PPARα. Our study suggests that in transiently transfected COS-1 cells, PFOA activate mouse and human PPARα, and PFDA activates mouse PPAR¿. This assay has the potential to evaluate and compare PPAR activation by members of the PFAA family of compounds. This abstract does not necessarily reflect USEPA policy.