Cyp1A and ABC transporter expression and function in PLHC-1cell line after microcystin-LR and Benzo (a)pyrene exposure

Name: Cyp1A and ABC transporter expression and function in PLHC-1cell line after microcystin-LR and Benzo (a)pyrene exposure
License: https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Keywords: Cytochrome p450 enzyme system

Datos de investigación

Cyp1A and ABC transporter expression and function in PLHC-1cell line after microcystin-LR and Benzo (a)pyrene exposure

Autores: Bieczynski, Flavia Icon

; Celander, M.C.; Edenius, M.; Painefilú, Julio César Icon

; Venturino, Andres Icon

; Luquet, Carlos Marcelo Icon

; Lindkvist, A.

Publicador: Consejo Nacional de Investigaciones Científicas y Técnicas

Fecha de depósito: 14/04/2025

Fecha de creación: 08-2024

Clasificación temática:

Bioquímica y Biología Molecular

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).

Palabras clave: Cytochrome p450 enzyme system, cytotoxicity, ABC transporter, microcystin-LR, ethoxy resorufin-O-deethylase activity, qPCR, polycyclic aromatic hydrocarbon, Poeciliopsis lucida hepatocellular carcinoma cell line

Previsualización destacada

Identificador del recurso

URI: http://hdl.handle.net/11336/258662

Colecciones

Datos de Investigación(CITAAC)
Datos de Investigación de CENTRO DE INVESTIGACIONES EN TOXICOLOGIA AMBIENTAL Y AGROBIOTECNOLOGIA DEL COMAHUE

Datos de Investigación(INIBIOMA)
Datos de Investigación de INST. DE INVEST.EN BIODIVERSIDAD Y MEDIOAMBIENTE

Datos de Investigación(IPATEC)
Datos de Investigación de INSTITUTO ANDINO PATAGONICO DE TECNOLOGIAS BIOLOGICAS Y GEOAMBIENTALES

Citación

Bieczynski, Flavia; Celander, M.C.; Edenius, M.; Painefilú, Julio César; Venturino, Andres; Luquet, Carlos Marcelo; Lindkvist, A.; (2025): Cyp1A and ABC transporter expression and function in PLHC-1cell line after microcystin-LR and Benzo (a)pyrene exposure. Consejo Nacional de Investigaciones Científicas y Técnicas. (dataset). http://hdl.handle.net/11336/258662

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Excepto donde se diga explícitamente, este item se publica bajo la siguiente descripción: Creative Commons Attribution-NonCommercial-ShareAlike 2.5 Unported (CC BY-NC-SA 2.5)