title: Expression of the selectable marker gene bsrm in BALB/MK cells induces apoptosis by overproduction of hydrogen peroxide.
abstract: Transduction of the retroviral vector LBmSN, which expresses the blasticidin S resistance gene bsrm in the murine keratinocyte cell line BALB/MK, induces death in these cells. Cell death is caused by a factor called DOKEB (death factor obtained from keratinocytes expressing bsrm), which is released before the cells' death. In this report we describe and discuss the purification and characterization of DOKEB. Our results were as follows. (i) The 5-day-old medium from the modified BALB/MK cells with LBmSN was used for purification and characterization by filtration and chromatography: DOKEB was a stable and highly hydrophilic compound, with a molecular mass less than that of 1 amino acid. (ii) The conditioned medium containing DOKEB was reactive against thiobarbituric acid and dichlorofluorescein diacetate. (iii) DOKEB activity was neutralized by the incubation of the conditioned medium with catalase. Therefore, our conclusion is that the BALB/MK cells expressing bsrm produce a large amount of hydrogen peroxide, which catalyzes the process of apoptosis of those cells.
authors: Department of Biophysics, UNIFESP, São Paulo-SP, Brazil.
authors: Daniela
authors: D
authors: Takeshita
authors: Fernanda Mara
authors: FM
authors: Bento
authors: Roger
authors: R
authors: Chammas
authors: José Ernesto
authors: JE
authors: Belizário
authors: Adriana Karaoglanovic
authors: AK
authors: Carmona
authors: Katsuhiro
authors: K
authors: Konno
authors: Robson
authors: R
authors: Lopes de Melo
authors: Gustavo
authors: G
authors: Molina
authors: Bianca Cristina Garcia
authors: BC
authors: Lisboa
authors: Sang Won
authors: SW
authors: Han
mesh_terms: D000619:Aminohydrolases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000302:isolation & purification / Q000378:metabolism; D016923:Cell Death; D002460:Cell Line; D004351:Drug Resistance / Q000235:genetics; D005819:Genetic Markers; D005822:Genetic Vectors / Q000235:genetics; D006861:Hydrogen Peroxide / Q000378:metabolism*; D015603:Keratinocytes / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic
chemical_list: D051017:Apoptosis Regulatory Proteins; D005819:Genetic Markers; D006861:Hydrogen Peroxide; D000619:Aminohydrolases; C019795:blasticidin S deaminase