Aziridinyl steroid-induced lesions in DNA and apoptosis in promyelocytic leukemia cells.
Variety of synthetic steroids are reported to be mutagenic as well as carcinogenic. The mutagenic and carcinogenic nature of these compounds have been related to their potential of being reactive to genetic material and production of reactive oxygen species. Here we have analyzed the action of aziridinyl steroid on calf thymus DNA and human promyelocytic leukemia cell line HL-60 under in vitro conditions. Calf thymus DNA when treated with various doses of aziridinyl steroid induced a high degree of stand separation and sensitivity/susceptibility to S1 nuclease hydrolysis. The treatment also induced an increasing number of strand breaks per molecule of DNA as determined by alkaline unwinding assay. Relatively higher doses of steroid, however, displayed a reduced susceptibility to S1 nuclease hydrolysis and did not increase the number of strand breaks in DNA. Moreover, the high dose treatments result increased melting temperature and an enhanced rate of reanealing after thermal denaturation, indicating that interstrand crosslinks are induced at higher doses of steroid treatment. Moreover, steroid treatment caused cell death in human promyelocytic leukemia cell line HL-60 and induced DNA degradation, characteristic of apoptosis. The test steroid has the ability to produce reactive oxygen intermediates (ROI) as determined by chemical methods. Incorporation of oxygen radical scavengers into the system blocked the damaging effect of steroid in calf thymus DNA and HL-60 cells. These observations strongly suggest that aziridinyl steroid, a pharmaceutical, damages mammalian DNA and induces apoptosis by the production of ROI in the test system.
Quadri
S A
SA
Department of Environmental Health, Boston University Medical Center, MA 02118, USA. qshafat@bu.edu
Islam
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S
Qadri
A N
AN
Ahmad
M
M
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001388:Aziridines / Q000633:toxicity*; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002777:Cholestanols / Q000633:toxicity*; D004247:DNA / Q000187:drug effects / Q000378:metabolism; D004249:DNA Damage; D004273:DNA, Neoplasm / Q000187:drug effects* / Q000378:metabolism*; D018922:HL-60 Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D006868:Hydrolysis; D009153:Mutagens / Q000633:toxicity*; D017382:Reactive Oxygen Species; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism
D001388:Aziridines; D002777:Cholestanols; D004273:DNA, Neoplasm; D009153:Mutagens; D017382:Reactive Oxygen Species; C071679:3-acetoxycholestanol(5,6-b)(N-phthalimido)aziridine; D004247:DNA; D006861:Hydrogen Peroxide; D015719:Single-Strand Specific DNA and RNA Endonucleases