Amplification of C1027-induced DNA cleavage and apoptosis by a quinacrine-netropsin hybrid molecule in tumor cell lines.
We examined the effect of a newly synthesized DNA-binding ligand, quinacrine-netropsin hybrid molecule (QN), on cytotoxicity, apoptosis, and DNA strand breaks induced by an enediyne antitumor antibiotic, C1027. QN significantly enhanced C1027-induced cellular DNA strand breaks, caspase-3 activation, and DNA ladder formation, characteristic of apoptosis, in human HL-60 cells. Flow cytometry revealed that C1027-induced intracellular H(2)O(2) generation was enhanced by QN, suggesting that QN enhances C1027-induced cytotoxic effect through H(2)O(2)-mediated apoptosis. QN also significantly enhanced C1027-induced apoptosis in BJAB cells, and the inhibition of apoptosis was observed in BJAB cells transfected with Bcl-2 gene. The experiment using (32)P-labeled DNA fragments showed that the addition of QN enhanced C1027-induced double-stranded DNA cleavage at the 5'-AGG-3'/3'-TCC-5' sequence (cutting sites are underlined). These results suggest that QN enhances C1027-induced antitumor effect via DNA cleavage and apoptosis. The present study shows a novel approach to the potentially effective anticancer therapy.
Iwamoto
Takuya
T
Department of Environmental and Molecular Medicine, Mie University School of Medicine, 2-174 Edobashi, Tsu, Mie 514 8507, Japan.
Hiraku
Yusuke
Y
Kojima
Michio
M
Kawanishi
Shosuke
S
D000617:Aminoglycosides / Q000494:pharmacology*; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis; D001665:Binding Sites; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival; D004247:DNA / Q000737:chemistry; D004249:DNA Damage; D053938:DNA Fragmentation; D053281:Enediynes; D004789:Enzyme Activation; D005434:Flow Cytometry; D018922:HL-60 Cells; D006801:Humans; D008024:Ligands; D008956:Models, Chemical; D009429:Netropsin / Q000494:pharmacology*; D009707:Nucleosomes / Q000378:metabolism; D010545:Peroxides / Q000737:chemistry; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D016283:Proto-Oncogene Proteins p21(ras) / Q000378:metabolism; D011796:Quinacrine / Q000494:pharmacology*; D013997:Time Factors; D014162:Transfection
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D000903:Antibiotics, Antineoplastic; D000970:Antineoplastic Agents; D053281:Enediynes; D008024:Ligands; D009707:Nucleosomes; D010545:Peroxides; D019253:Proto-Oncogene Proteins c-bcl-2; C058024:C 1027; D009429:Netropsin; D004247:DNA; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras); D011796:Quinacrine