Cyp1A and ABC transporter expression and function in PLHC-1cell line after microcystin-LR and Benzo (a)pyrene exposure
; Celander, M.C. ; Edenius, M. ; Painefilú, Julio César ; Venturino, Andres ; Luquet, Carlos Marcelo ; Lindkvist, A. Otros responsables
Fecha
Publicador:
Consejo Nacional de Investigaciones Científicas y Técnicas
Fecha de depósito:
14/04/2025
Fecha de creación:
08/2024
Clasificación temática
Resumen
The dataset presented corresponds to a study investigating mixture toxicity between two common aquatic contaminants, microcystin-LR (MCLR) and benzo[a]pyrene (BaP), on the detoxification system of fish. We used the Poeciliopsis lucida hepatocellular carcinoma (PLHC-1) cell line as a model for fish liver cells. The data was collected from May 2022 to August 2024. Cells were exposed to MCLR (0.01, 1 µM), BaP (0.01, 0.1, 1 µM), or combinations of both chemicals for periods ranging from 1 to 48 hours. We measured the following endpoints: - Cytochrome P450 1A (CYP1A) function and regulation, assessing ethoxy resorufin-O-deethylase (EROD) activity and CYP1A mRNA expression. EROD activity was normalized to the protein. - P-glycoprotein (Pgp) function and expression, using the rhodamine 123 (Rh123) accumulation assay and Pgp mRNA expression. Rh123 accumulation was normalized to the protein. - Cell cytotoxicity, using two fluorescent indicators, 5-Carboxyfluorescein Diacetate-Acetoxymethyl Ester (CFDA-AM) and Alamar Blue (AB).
Métodos
1) CYP1A Activity: This section contains raw data used to calculate CYP1A activity, expressed as specific EROD activity. The EROD assay is based on the ability of the CYP1A enzyme to catalyze the O-deethylation of ethoxyresorufin to resorufin, a fluorescent product. Resorufin production was monitored over time and quantified using a resorufin standard curve. Each sample value was normalized to total protein content, which was measured using fluorescamine, and bovine serum albumin (BSA) as the protein standard. The files include: - EROD slopes for each sample - Protein concentration per sample - Specific EROD activity - Standard curves (for BSA and resorufin). Additionally, the dataset includes results from an experiment involving pre-induction of EROD activity using β-naphthoflavone (BNF). Detailed descriptions of measurement conditions and calculation methods are provided in the accompanying README file (EROD_readme).
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