title: Characteristics of sulfasalazine-induced cytotoxicity in C6 rat glioma cells.
abstract: Glioblastoma is the most aggressive primary brain tumor. Surgical resection, radiotherapy and temozolomide (TMZ), an alkylating agent, is the standard of care. Glioma cells may synthetize the antioxidant glutathione by importing cystine through a cystine/glutamate antiporter, which is inhibited by sulfasalazine (SAS). C6 rat glioma cells are largely used in in vitro and in vivo models for developing new glioblastoma treatment strategies. We treated C6 cells with 25μM TMZ and/or 0.25mM or 0.5mM SAS for 1, 3 or 5days and evaluated viability, apoptosis, total glutathione levels and metalloproteinase MMP2 and MMP9 activities. TMZ treatment slightly reduced cell viability by 9.5% compared with vehicle treatment (0.1% dimethyl sulfoxide) only after 5days. In addition, TMZ did not modify apoptosis, glutathione content or MMP2/MMP9 activities. The 0.25mM SAS treatment reduced cell viability by 31.1% and 19.4% after the first and third days, respectively. This effect was not sustained after the fifth day of treatment. In contrast, 0.5mM SAS caused a reduction in cell viability by nearly 100%, total glutathione depletion and apoptosis induction. Moreover, the effect of 0.5mM SAS was greater than that of TMZ in terms of cell viability reduction, total glutathione depletion and apoptosis induction. MMP9 activity was reduced by 40% after 5days of 25μM TMZ and 0.5mM SAS co-administration. Considering previous data from our group, we verified that the cellular viability results differed between rat and human cells; C6 cells were more vulnerable to 0.5mM SAS than human A172 and T98G glioblastoma lineages. We propose that C6 cells may not be appropriate for studying human glioblastoma and that the results obtained using these cells should be interpreted with caution.
authors.affiliation: Department of Anatomical Pathology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Av. Tessália Vieira de Camargo, 126, 13083-887, Campinas, SP, Brazil.
authors.forename: Raffaela Silvestre
authors.initials: RS
authors.lastname: Ignarro
authors.affiliation: Department of Anatomical Pathology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Av. Tessália Vieira de Camargo, 126, 13083-887, Campinas, SP, Brazil.
authors.forename: Gustavo
authors.initials: G
authors.lastname: Facchini
authors.affiliation: Department of Clinical Pathology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Av. Tessália Vieira de Camargo, 126, 13083-887, Campinas, SP, Brazil.
authors.forename: Daniela Rodrigues
authors.initials: DR
authors.lastname: de Melo
authors.affiliation: Department of Biochemistry and Tissue Biology, Institute of Biology, Laboratory of Bioassays and Signal Transduction, State University of Campinas (UNICAMP), Street Monteiro Lobato, 255, 13083-970, Campinas, SP, Brazil.
authors.forename: Karin Juliane
authors.initials: KJ
authors.lastname: Pelizzaro-Rocha
authors.affiliation: Department of Biochemistry and Tissue Biology, Institute of Biology, Laboratory of Bioassays and Signal Transduction, State University of Campinas (UNICAMP), Street Monteiro Lobato, 255, 13083-970, Campinas, SP, Brazil.
authors.forename: Carmen Veríssima
authors.initials: CV
authors.lastname: Ferreira
authors.affiliation: Department of Clinical Pathology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Av. Tessália Vieira de Camargo, 126, 13083-887, Campinas, SP, Brazil.
authors.forename: Roger Frigério
authors.initials: RF
authors.lastname: Castilho
authors.affiliation: Department of Anatomical Pathology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Av. Tessália Vieira de Camargo, 126, 13083-887, Campinas, SP, Brazil. Electronic address: fabio@fcm.unicamp.br.
authors.forename: Fabio
authors.initials: F
authors.lastname: Rogerio
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003606:Dacarbazine / Q000031:analogs & derivatives / Q000494:pharmacology; D005910:Glioma; D005978:Glutathione / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051381:Rats; D012460:Sulfasalazine / Q000494:pharmacology*; D000077204:Temozolomide
chemical_list: D000970:Antineoplastic Agents; D012460:Sulfasalazine; D003606:Dacarbazine; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D005978:Glutathione; D000077204:Temozolomide
keywords: Apoptosis; C6 lineage; Glioblastoma model; Glutathione; Viability