Cytotoxicity of RH1: NAD(P)H:quinone acceptor oxidoreductase (NQO1)-independent oxidative stress and apoptosis induction.
The elevated expression of the flavoprotein NAD(P)H:quinone acceptor oxidoreductase (NQO1) (EC 1.6.99.2) in many human solid tumors, along with its ability to activate quinone-based anticancer agents, makes it an excellent target for enzyme-directed drug development. Previous studies have shown a significant statistical correlation between NQO1 enzymatic activity and the cytotoxicity of certain antitumor quinones. RH1 [2,5-diaziridinyl-3-(hydroxymethyl)-6-methyl-1,4-benzoquinone], presently in late preclinical and entering early clinical development, has been previously considered to be an excellent substrate for activation by NQO1. In this study we investigate the cytotoxicity of RH1 in cell lines selected from the NCI's 60 tumor cell line panel, expressing varying levels of NQO1 activity. Exposure time- and concentration-dependent cytotoxicity was seen, apparently independent from levels of NQO1 activity in these cells. Furthermore, the NQO1 inhibitor dicoumarol had no impact on the sensitivity profiles of RH1 response. The HL-60 myeloid leukemia cells, which do not have detectable NQO1 activity, were further investigated. RH1 treatment of HL-60 cells generated high levels of free radicals, which was accompanied by robust redox cycling, oxygen consumption and induction of apoptosis. These results are in agreement with previous data suggesting that, in addition to its activation by NQO1, RH1-induced cytotoxicity might involve alternative pathways for activation of this compound. Furthermore, the high cytotoxicity of RH1 in the leukemia/lymphoma subpanel of the NCI in vitro cell line screen would suggest an empirical rationale for the utilization of this compound in the treatment of these malignancies.
Tudor
Gabriela
G
Science Applications International Corp., National Cancer Institute, Frederick, MD, USA.
Alley
Mike
M
Nelson
Christopher M
CM
Huang
Ruili
R
Covell
David G
DG
Gutierrez
Peter
P
Sausville
Edward A
EA
D017209:Apoptosis / Q000187:drug effects*; D001388:Aziridines / Q000633:toxicity*; D016227:Benzoquinones / Q000633:toxicity*; D004305:Dose-Response Relationship, Drug; D005609:Free Radicals / Q000378:metabolism; D018922:HL-60 Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D016660:NAD(P)H Dehydrogenase (Quinone) / Q000378:metabolism*; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000187:drug effects*; D010100:Oxygen / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
C117776:2,5-diaziridinyl-3-(hydroxymethyl)-6-methyl-1,4-benzoquinone; D001388:Aziridines; D016227:Benzoquinones; D005609:Free Radicals; D016660:NAD(P)H Dehydrogenase (Quinone); C447479:NQO1 protein, human; D010100:Oxygen