title: Serum depletion, multidrug resistance and fluorescent probes: methodological implications in free radicals evaluation.
abstract: Intracellular probes used for oxidative burst evaluation could be a substrate of the multidrug resistance proteins (MDR), which may cause misinterpretation of experimental data. We aimed to study the effect of the culture condition and of the MDR-interfering antioxidant quercetin on free radical measurement in serum depleted HCT-8 cells and chick embryo hepatocytes. Serum depletion and/or quercetin affected the traffic of 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein (BCECF) and calcein. The comparative results obtained with 2',7'-dichlorofluorescein (DCF) and with the plasma-membrane probe C11-BODIPY 581/591 [4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s-indacene-3-undecanoic acid], suggest caution in the use of intracellular probes to evaluate oxidative stress in vitro.
authors.affiliation: Department of Physiology and Pharmacology "V. Erspamer", "Sapienza" University of Rome, Italy.
authors.forename: Ilaria
authors.initials: I
authors.lastname: Peluso
authors.affiliation: Department of Physiology and Pharmacology "V. Erspamer", "Sapienza" University of Rome, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Romanelli
authors.affiliation: Department of Physiology and Pharmacology "V. Erspamer", "Sapienza" University of Rome, Italy.
authors.forename: Silvana
authors.initials: S
authors.lastname: Spagnuolo
authors.affiliation: Department of Physiology and Pharmacology "V. Erspamer", "Sapienza" University of Rome, Italy.
authors.forename: Husseen
authors.initials: H
authors.lastname: Manafikhi
authors.affiliation: Department of Physiology and Pharmacology "V. Erspamer", "Sapienza" University of Rome, Italy. Electronic address: maura.palmery@uniroma1.it.
authors.forename: Maura
authors.initials: M
authors.lastname: Palmery
mesh_terms: D000578:Amidines / Q000494:pharmacology; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D001896:Boron Compounds / Q000378:metabolism / Q000493:pharmacokinetics; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D002642:Chick Embryo; D002645:Chickens; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D018432:Drug Resistance, Multiple; D005452:Fluoresceins / Q000378:metabolism / Q000493:pharmacokinetics; D005456:Fluorescent Dyes / Q000378:metabolism* / Q000493:pharmacokinetics; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D018384:Oxidative Stress / Q000187:drug effects; D011794:Quercetin / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods
chemical_list: D000578:Amidines; D001896:Boron Compounds; C120421:C11-BODIPY(581-591); D016895:Culture Media, Serum-Free; D005452:Fluoresceins; D005456:Fluorescent Dyes; D017382:Reactive Oxygen Species; C037631:2',7'-dichlorofluorescein; C046728:2,2'-azobis(2-amidinopropane); C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein; D011794:Quercetin; C007740:fluorexon
keywords: Intracellular probes; Multidrug resistance; Oxidative stress; Quercetin