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Martina Stepankova DOI: 10.3252/pso.eu.18ECE.2016 Nuclear receptors and Signal transduction 2. Effects of statin enantiomers on transcriptional activity of PXR in LS180 cells. Prior to gene reporter assays, we examined the cytotoxicity of tested compounds in human colon adenocarcinoma LS180 cell line. For this purpose, the cells were incubated for 24 h with individual enantiomers of atorvastatin, fluvastatin and rosuvastatin at concentrations ranging from 100 pM to 100 μM. The vehicle was DMSO (0.1% v/v). The data are the mean ± SD from experiments performed in three consecutive passages of cells and are expressed as percentage of viability of control cells. The values of IC50 were calculated where appropriate and they are indicated in plots (Figure 2, upper panels). Transcriptional activity of PXR was tested in LS180 cells transiently transfected with p3A4luc reporter construct, incubated for 24 h with individual statin enantiomers in concentration ranging from 100 pM to 100 μM in the absence (agonist mode – middle panels) or presence (antagonist mode – lower panels) of rifampicin (RIF; 10 μM). The vehicle was DMSO (0.1% v/v). After the treatments, cells were lysed and luciferase activity was measured. Treatments were performed in triplicates. Data are expressed as a fold induction of luciferase activity over control cells (agonist mode) or as a percentage of maximal activation attained by RIF (antagonist mode). The values of EC50 and IC50 from n independent cell passages were calculated where appropriate and the average values are indicated in plots. Representative gene reporter assays are shown.

Optical isomers of statins enantiospecifically activate pregnane X receptor PXR and induce CYP2A6, CYP2B6 and CYP3A4 in human hepatocytes