title: In vitro study of putative genomic biomarkers of nephrotoxicity through differential gene expression using gentamicin.
abstract: Drug-induced nephrotoxicity is one of the most frequently observed effects in long-term pharmacotherapy. The effects of nephrotoxicity are commonly discovered later due to a lack of sensitivity in in vivo methods. Therefore, researchers have tried to develop in vitro alternative methods for early identification of toxicity. In this study, LLC-PK1 cells were exposed to gentamicin through MTT and trypan blue assay. Concentrations of 4 (low), 8 (medium) and 12 (high) mM, were used to evaluate differential gene expression. A panel of genes was selected based on gene expression changes. The search for sequences of mRNA encoding proteins previously associated with kidney damage was conducted in the databases of the National Center for Biotechnology Information (USA). RNA was extracted from the cells, and RT-qPCR was performed to evaluate differential expression profiles of the selected genes. Among the 11 analyzed genes, four proved to be differentially up-regulated in cells exposed to gentamicin: HAVcr1, caspase 3, ICAM-1 and EXOC6. According to this study's results, we suggest that these genes play an important role in the mechanism of in vitro nephrotoxicity caused by gentamicin and can be used as early in vitro biomarkers to identify nephrotoxicity when developing safer drugs.
authors.affiliation: a Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, Laboratório de Toxicologia Experimental in vitro , Universidade Federal de Minas Gerais (UFMG) , Belo Horizonte , Brazil.
authors.forename: Sarah Cristina Teixeira
authors.initials: SCT
authors.lastname: Silva
authors.affiliation: b Departamento de Microbiologia e Imunologia, Laboratório de Imunologia, Instituto de Ciências Biomédicas , Universidade Federal de Alfenas (UniFal) , Alfenas , Brazil.
authors.forename: Leonardo Augusto
authors.initials: LA
authors.lastname: de Almeida
authors.affiliation: a Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, Laboratório de Toxicologia Experimental in vitro , Universidade Federal de Minas Gerais (UFMG) , Belo Horizonte , Brazil.
authors.forename: Stellamaris
authors.initials: S
authors.lastname: Soares
authors.affiliation: a Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, Laboratório de Toxicologia Experimental in vitro , Universidade Federal de Minas Gerais (UFMG) , Belo Horizonte , Brazil.
authors.forename: Marina Felipe
authors.initials: MF
authors.lastname: Grossi
authors.affiliation: c Departamento de Nutrição , Universidade Federal de Juiz de Fora (UFJF) - Campus Governador Valadares , Governador Valadares , Brazil.
authors.forename: Anete Maria Santana
authors.initials: AMS
authors.lastname: Valente
authors.affiliation: a Departamento de Análises Clínicas e Toxicológicas, Faculdade de Farmácia, Laboratório de Toxicologia Experimental in vitro , Universidade Federal de Minas Gerais (UFMG) , Belo Horizonte , Brazil.
authors.forename: Carlos Alberto
authors.initials: CA
authors.lastname: Tagliati
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D020869:Gene Expression Profiling; D005819:Genetic Markers; D005839:Gentamicins / Q000633:toxicity*; D007668:Kidney / Q000187:drug effects* / Q000378:metabolism; D018374:LLC-PK1 Cells; D009153:Mutagens / Q000633:toxicity*; D012333:RNA, Messenger / Q000378:metabolism; D013552:Swine; D059467:Transcriptome / Q000187:drug effects*; D015854:Up-Regulation
chemical_list: D005819:Genetic Markers; D005839:Gentamicins; D009153:Mutagens; D012333:RNA, Messenger
keywords: Nephrotoxicity; genomic biomarkers; gentamicin