Growth inhibition and apoptosis induced by 2-phenoxymethyl-3H-quinazolin-4-one in HL-60 leukemia cells.
OBJECTIVE
The aim of the study was to investigate anticancer activity of newly synthesized 2-phenoxymethyl-3H-quinazolin-4-one (PMQ).


METHODS
Anticancer activity of PMQ was studied towards human HL-60 leukemia cells. Antiproliferative activity of PMQ was determined by direct counting of cells using trypan blue staining technique. Apoptosis and cell cycle profile changes were analysed using internucleosomal DNA fragmentation assay and flow cytometry. Activation of caspases and changes in glutathione level were monitored using colorimetric or luminiscent methods.


RESULTS
PMQ induced concentration-dependent cytotoxicity in leukemia cells, with IC(50) of 10.8 +/- 0.9 microM. DNA flow cytometry analysis and DNA ladder formation assay indicated that PMQ actively induced apoptosis of cells accompanied by a block of cells in G(2)/M phase and a marked loss of cells in G(0)/G(1) and S phases. Additionally, the activities of caspase-3 and caspase-9 were increased significantly and a markedly increased level of oxidized glutahione was observed. Inhibition of glutahione synthesis using buthionine sulfoximine sensitized leukemia cells to PMQ, confirming the involvement of ROS in PMQ-induced apoptosis.


CONCLUSIONS
The results of this study clearly demonstrate that PMQ is a promising anticancer drug showing cytostatic and apoptotic effects toward HL-60 leukemia cells mainly through mitochondrial/caspase-9 dependent pathway.
Cipak
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Cancer Research Institute, Slovak Academy of Sciences, 833 91 Bratislava, Slovakia. exoncip@savba.sk
Repicky
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Jantova
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D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D005978:Glutathione / Q000378:metabolism; D018922:HL-60 Cells / Q000187:drug effects; D006801:Humans; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2; D011799:Quinazolines / Q000737:chemistry / Q000494:pharmacology*; D017382:Reactive Oxygen Species / Q000378:metabolism
C520588:2-phenoxymethyl-3H-quinazolin-4-one; D019253:Proto-Oncogene Proteins c-bcl-2; D011799:Quinazolines; D017382:Reactive Oxygen Species; D053148:Caspase 3; D053453:Caspase 9; D005978:Glutathione