title: Ribonuclease binase apoptotic signature in leukemic Kasumi-1 cells.
abstract: Cytotoxic exogenous RNases triggering apoptotic response in malignant cells have potential as anticancer drugs; surprisingly, detailed characterization of the RNase-induced apoptosis has not been conducted so far. Here we show that a cytotoxic RNase from Bacillus intermedius (binase) induces extrinsic and intrinsic apoptotic pathways in leukemic Kasumi-1 cells. The experiments were performed using TaqMan Array Human Apoptosis 96-well Plate for gene expression analysis, and flow cytometry. Cytometric studies demonstrated dissipation of the mitochondrial membrane potential, opening of mitochondrial permeability transition pores, activation of caspases, increase of intracellular Ca(2+) and decrease of reactive oxygen species levels. We found that expression of 62 apoptotic genes is up-regulated, including 16 genes that are highly up-regulated, and only one gene was found to be down-regulated. The highest, 16 fold increase of the expression level was observed for TNF gene. Highly up-regulated genes also include the non-canonical NF-κB signaling pathway and inflammatory caspases 1,4. The obtained results suggest that binase induces evolutionary acquired cellular response to a microbial agent and triggers unusual apoptosis pathway.
authors.affiliation: Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, Vavilova Str 32, 119991 Moscow, Russia.
authors.forename: Vladimir A
authors.initials: VA
authors.lastname: Mitkevich
authors.forename: Olga V
authors.initials: OV
authors.lastname: Kretova
authors.forename: Irina Yu
authors.initials: IY
authors.lastname: Petrushanko
authors.forename: Ksenia M
authors.initials: KM
authors.lastname: Burnysheva
authors.forename: Dmitry V
authors.initials: DV
authors.lastname: Sosin
authors.forename: Olga V
authors.initials: OV
authors.lastname: Simonenko
authors.forename: Olga N
authors.initials: ON
authors.lastname: Ilinskaya
authors.forename: Nickolai A
authors.initials: NA
authors.lastname: Tchurikov
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Makarov
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D004722:Endoribonucleases / Q000378:metabolism / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D033681:Mitochondrial Membrane Transport Proteins / Q000187:drug effects / Q000378:metabolism; D000083162:Mitochondrial Permeability Transition Pore; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059467:Transcriptome / Q000187:drug effects*
chemical_list: D033681:Mitochondrial Membrane Transport Proteins; D000083162:Mitochondrial Permeability Transition Pore; D004722:Endoribonucleases; C022543:ribonuclease T(2)