We investigate the roles of methoxyl (OCH(3)) and
hydroxyl (
OH) substitutions at C8 of
flavonoids on their apoptosis-inducing activities.
Wogonin (Wog) and
nor-wogonin (N-Wog) are structurally related
flavonoids, and respectively contain an
OH and OCH(3) at C8. In
leukemia HL-60 cells, N-Wog exhibited more-potent cytotoxicity than Wog according to the MTT and LDH release assays, and the IC(50) values of Wog and N-Wog in HL-60 cells were 67.5 +/- 2.1 and 21.7 +/- 1.5 microM, respectively. Apoptotic characteristics including
DNA ladders, apoptotic bodies, and hypodiploid cells accompanied by the induction of
caspase 3 protein processing appeared in Wog- and N-Wog-treated HL-60 cells. Interestingly, an increase in intracellular
peroxide production was detected in N-Wog- but not Wog-treated HL-60 cells by the DCHF-DA assay, and the reduction of intracellular
peroxide by
catalase (CAT) induced by N-Wog significantly reduced the N-Wog- but not the Wog-induced cytotoxic effect according to the MTT assay in accordance with the blocking of
DNA ladder formation and
caspase 3 and PARP
protein processing elicited by N-Wog. We further analyzed the effect of six structurally related compounds, including 5-OH, 7-OH, 5,7-diOH, 5,7-diOCH(3), 7,8-diOCH(3), and 7-OCH(3)-8-OH
flavones, on apoptosis induction in HL-60 cells. Results suggested that
OH at C5 and C7 is essential for both the apoptosis-inducing activity of
flavonoids, and
OH at C8 may contribute to apoptosis induction ability. Evidence to support a distinct structure-activity relationship in apoptosis induction of
flavonoids is provided for the first time in this study.