A role for apoptosis in the toxicity and mutagenicity of bleomycin in AHH-1 tk+/- human lymphoblastoid cells.
The chromosomal mutagen, bleomycin, is also noted for its toxic properties, although the mechanism of cell death is not fully understood. In order to determine if cell death occurred by apoptosis or necrosis, AHH-1 tk+/- cells were exposed to bleomycin and the percentage of viable, apoptotic and necrotic cells quantified by flow cytometry. Logistic regression analysis indicated that the primary manner of cell death was through the apoptosis pathways, that apoptosis was delayed, and that apoptosis was accompanied by an arrest in the G2 phase of the cell cycle. Once apoptosis was established as a mechanism for cell death, the efficiency with which these pathways removed damaged cells from the population was evaluated with the use of specific-locus mutation assays (tk and hprt) as indicators of cells with DNA damage that maintained viability and clonogenicity. Linear regression analysis detected a significant, concentration-dependent increase in the numbers of TFTr clones with the slow-growth phenotype. This suggests that a proportion of cells with bleomycin-induced DNA damage did not undergo cell death by apoptosis and that apoptosis, a mechanism for the destruction of damaged cells, is not fully efficient in the AHH-1 tk +/- cell line.
Morris
S M
SM
Department of Health and Human Services, Food and Drug Administration, Jefferson, AR 72079, USA. smorris@nctr.fda.gov
Domon
O E
OE
McGarrity
L J
LJ
Chen
J J
JJ
Manjanatha
M G
MG
Andrews
A M
AM
Aidoo
A
A
Casciano
D A
DA
D017209:Apoptosis / Q000187:drug effects*; D001402:B-Lymphocytes / Q000187:drug effects; D001761:Bleomycin / Q000633:toxicity*; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004249:DNA Damage; D006801:Humans; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D016296:Mutagenesis / Q000187:drug effects; D009153:Mutagens / Q000633:toxicity*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D013937:Thymidine Kinase / Q000378:metabolism
D009153:Mutagens; D019253:Proto-Oncogene Proteins c-bcl-2; D001761:Bleomycin; D013937:Thymidine Kinase