title: Cytotoxic effect of amphotericin B in a myofibroblast cell line.
abstract: In this study we investigate whether Amphotericin B (AmB), a widely used antifungal agent, could decrease the proliferation of a myofibroblast cell line - GRX, a model of activated hepatic stellate cells (HSC). Three different hepatic cell lines (GRX, Hep G2 and ARL-6) were treated with two concentrations of AmB (1.25 μg/mL or 2.50 μg/mL). Cytotoxicity was assessed by MTT assay. The effects of AmB on GRX migration was evaluated by Wound-healing Assay. Cell cycle arrest was investigated by flow cytometry. Apoptosis and autophagy were analyzed by Caspase 3 and LC3 immunostaining, respectively. Treatment with AmB 1.25 or 2.50 μg/mL showed a decrease in viability of GRX cells. This decrease was not observed for Hep G2 or ARL-6 in any of the two AmB concentrations tested. GRX cells treated with 1.25 μg/mL AmB were unable to close the wound after 96 h. Cell cycle analysis showed an increase in sub-G1 population and a decrease in G2/M population in AmB-treated cells. In addition, AmB-treated GRX cells showed increased expression of LC-3 and Caspase-3 by immunohistochemistry, suggesting an increase in both autophagy and apoptosis. Here we show that AmB is cytotoxic for GRX cells, a model of activated HSC, but not for hepatic lineages HepG2 and ARL6.
authors.affiliation: Gene Therapy Center, Experimental Research Center, Hospital de Clinicas de Porto Alegre, Porto Alegre, RS 90035-903, Brazil; Post Graduation Program in Genetics and Molecular Biology, UFRGS, Porto Alegre, RS 91501-970, Brazil. Electronic address: carolinaurib10@yahoo.com.ar.
authors.forename: C C
authors.initials: CC
authors.lastname: Uribe
authors.forename: F
authors.initials: F
authors.lastname: Dos Santos de Oliveira
authors.forename: B
authors.initials: B
authors.lastname: Grossmann
authors.forename: N A
authors.initials: NA
authors.lastname: Kretzmann
authors.forename: T
authors.initials: T
authors.lastname: Reverbel da Silveira
authors.forename: R
authors.initials: R
authors.lastname: Giugliani
authors.forename: U
authors.initials: U
authors.lastname: Matte
mesh_terms: D000666:Amphotericin B / Q000009:adverse effects*; D000818:Animals; D000935:Antifungal Agents / Q000009:adverse effects*; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D015415:Biomarkers / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016193:G1 Phase / Q000187:drug effects; D056945:Hep G2 Cells; D055166:Hepatic Stellate Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D051379:Mice; D058628:Myofibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009994:Osmolar Concentration
chemical_list: D000935:Antifungal Agents; D015415:Biomarkers; D000666:Amphotericin B
keywords: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltet-razoliumbromide; 4′,6-diamidino-2-phenylindole; AmB; Amphotericin B; Apoptosis; DAPI; DMEM; Dulbecco’s Modified Eagle’s Medium; ECM; GRX cells; HSC; Hepatic Stellate Cells; Hepatic stellate cells; MTT; PBS; Phosphate Buffered Saline; alpha smooth muscle actin; extracellular matrix; α-SMA