caa a22 4500 60623473X CHVBK 20210128101238.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s11010-015-2518-8 doi (NATIONALLICENCE)springer-10.1007/s11010-015-2518-8 Dual properties of hispidulin: antiproliferative effects on HepG2 cancer cells and selective inhibition of ABCG2 transport activity [Elektronische Daten] [Carina Scoparo, Glaucio Valdameri, Paulo Worfel, Fernanda Guterres, Glaucia Martinez, Sheila Winnischofer, Attilio Di Pietro, Maria Rocha] Hepatocellular carcinoma is the third most common cause of cancer-related deaths worldwide. Furthermore, the existing pharmacological-based treatments are insufficiently effective and generate many side effects. Hispidulin (6-methoxy-5,7,4′-trihydroxyflavone) is a flavonoid found in various medicinal herbs that present antineoplastic properties. Here we evaluated how modulation of reactive oxygen species (ROS) and alterations of antioxidant defenses could be associated to the antiproliferative effects of hispidulin in HepG2 cells. In addition, we studied the inhibitory activity of hispidulin on the efflux of drugs mediated by ABC transporters involved in multidrug resistance. In order to understand the increase of intracellular ROS promoted by hispidulin, we investigated the mRNA expression levels and activities of antioxidant enzymes, and the GSH/GSSG ratio. We showed that hispidulin significantly down-regulated the transcription levels of catalase, leading to reduction of enzyme activity and decrease of the GSH content. We also observed that, in the presence of N-acetylcysteine or exogenous catalase, the proliferation was lowered back to the control levels. These data clearly indicate a strong involvement of intracellular ROS levels for triggering the antiproliferative effects. We also demonstrated that the inhibition produced by hispidulin on drug efflux was specific for ABCG2, since no effects were observed with ABCB1 and ABCC1. Furthermore, HepG2 cells were more sensitive to hispidulin-mediated cell death than immortalized L929 fibroblasts, suggesting a differential toxicity of this compound between tumor and non-tumor cell lines. Our results suggest that hispidulin constitutes a promising candidate to sensitize chemoresistant cancer cells overexpressing ABCG2. Springer Science+Business Media New York, 2015 Hispidulin nationallicence HepG2 cells nationallicence ROS nationallicence Catalase nationallicence ABCG2 nationallicence 7-AAD : (7-Amino-actinomycin D) nationallicence CAT : Catalase nationallicence DCFH-DA : 2′,7′-Dichlorofluorescin diacetate nationallicence DMSO : Dimethyl sulfoxide nationallicence FBS : Fetal bovine serum nationallicence Gred : Glutathione reductase nationallicence GSH : Glutathione nationallicence GPx : Glutathione peroxidase nationallicence GSSG : Glutathione disulfide nationallicence HCC : Hepatocellular carcinoma nationallicence H2O2 : Hydrogen peroxide nationallicence HCl : Hydrochloric acid nationallicence MTT : 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide nationallicence NAC : N-acetylcysteine nationallicence NADPH : Nicotinamide adenine dinucleotide phosphate nationallicence PBS : Phosphate buffered saline nationallicence PCR : Polymerase chain reaction nationallicence PE : Phycoerythrin nationallicence PI : Propidium iodide nationallicence ROS : Reactive oxygen species nationallicence SOD : Superoxide dismutase nationallicence Scoparo Carina Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut Valdameri Glaucio Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut Worfel Paulo Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut Guterres Fernanda Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut Martinez Glaucia Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut Winnischofer Sheila Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut Di Pietro Attilio Equipe Labellisée Ligue 2014, BMSSI UMR 5086 CNRS, IBCP, Université Lyon 1, Lyon, France aut Rocha Maria Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 409/1-2(2015-11-01), 123-133 0300-8177 409:1-2<123 2015 409 11010 https://doi.org/10.1007/s11010-015-2518-8 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-015-2518-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Scoparo Carina Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut NATIONALLICENCE 700 1- Valdameri Glaucio Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut NATIONALLICENCE 700 1- Worfel Paulo Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut NATIONALLICENCE 700 1- Guterres Fernanda Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut NATIONALLICENCE 700 1- Martinez Glaucia Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut NATIONALLICENCE 700 1- Winnischofer Sheila Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut NATIONALLICENCE 700 1- Di Pietro Attilio Equipe Labellisée Ligue 2014, BMSSI UMR 5086 CNRS, IBCP, Université Lyon 1, Lyon, France aut NATIONALLICENCE 700 1- Rocha Maria Department of Biochemistry and Molecular Biology, Federal University of Paraná, Curitiba-PR, Brazil aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 409/1-2(2015-11-01), 123-133 0300-8177 409:1-2<123 2015 409 11010