title: Novel nonclassical antifolate, 2-[N-(2´-Hydroxyethyl)amino]methyl-3H-quinazolin-4-one, with a potent antineoplastic activity toward leukemia cells.
abstract: This study was aimed to investigate the therapeutic potential of novel nonclassical antifolate, 2-[N-(2´-Hydroxyethyl)ami-no]methyl-3H-quinazolin-4-one (HEAMQ), toward human promonocytic U937 and murine lymphoblastic L1210 cell lines. The antiproliferative activity of HEAMQ was determined by MTT assay and its effects on cell cycle progression and apoptosis were studied by flow cytometry, and by immunoblots, respectively. In addition, combination chemotherapy of HEAMQ with cisplatin and temozolomide under in vitro and in vivo conditions was tested. HEAMQ showed concentration- and time-dependent cytotoxicity toward U937 and L1210 cells. It induced G2/M arrest that in U937 cells was associated with a marked decrease in the protein expressions of cyclin A, cyclin B, and cyclin-dependent kinase Cdk1. HEAMQ-induced apoptosis was accompanied with up-regulation of the protein expression of Bax and down-regulation of the protein expression of Bcl-2, Mcl-1, XIAP, and survivin, resulting in cytochrome c release and activation of caspases. Inhibitors of JNK (SP600125) and p38 MAPK (SB203580) suppressed HEAMQ-induced apoptosis and G2/M phase arrest, attenuated the activation of Bax, and blocked down-regulation of Bcl-2, XIAP and survivin in HEAMQ-treated U937 cells. In addition, combinations of HEAMQ with cisplatin and temozolomide resulted in synergistic inhibition of cell growth, producing long-term survivors of L1210 tumor-bearing mice. In conclusion, these results indicate that HEAMQ antineoplastic activity toward leukemia cells is associated with cell cycle arrest and apoptosis. The in vivo studies further confirmed the antitumor activity of HEAMQ and highlighted that this agent could be used to further increase therapeutic efficacies of traditional chemotherapeutic agents.
authors.affiliation: Cancer Research Institute, Slovak Academy of Sciences, Vlarska 7, 83391 Bratislava, Slovakia. exoncip@savba.sk
authors.forename: L
authors.initials: L
authors.lastname: Cipak
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology; D003606:Dacarbazine / Q000031:analogs & derivatives / Q000494:pharmacology; D005260:Female; D005493:Folic Acid Antagonists / Q000494:pharmacology*; D006801:Humans; D007938:Leukemia / Q000188:drug therapy*; D007939:Leukemia L1210; D008297:Male; D051379:Mice; D008811:Mice, Inbred DBA; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors; D011799:Quinazolines / Q000494:pharmacology*; D000077204:Temozolomide; D020298:U937 Cells
chemical_list: C577340:2-(N-(2-hydroxyethyl)amino)methyl-3H-quinazolin-4-one; D000970:Antineoplastic Agents; D005493:Folic Acid Antagonists; D011799:Quinazolines; D003606:Dacarbazine; D020928:Mitogen-Activated Protein Kinases; D002945:Cisplatin; D000077204:Temozolomide