Homocysteine thiolactone induces apoptotic DNA damage mediated by increased intracellular hydrogen peroxide and caspase 3 activation in HL-60 cells.
The cytotoxicity of homocysteine derivatives on chromosomal damage in somatic cells is not well established. The present study used reactive homocysteine derivative of homocysteine thiolactone (Hcy) to investigate its causal effect on apoptotic DNA injury in human promyeloid HL-60 cells. Our results demonstrated that Hcy induced cell death and features of apoptosis including increased phosphotidylserine exposure on the membrane surface, increased apoptotic cells with hypoploid DNA contents, and internucleosomal DNA fragmentation, all of which occurred in a time- and concentration-dependent manner. Hcy treatment also significantly increased intracellular reactive oxygen species H2O2, which coincided with the elimination of caspase 3 proenzyme levels and increased caspase 3 activity at the time of the appearance of apoptotic DNA fragmentation. Preincubation of Hcy-treated HL-60 cells with catalase completely scavenged intracellular H2O2, thus inhibiting caspase 3 activity and protecting cells from apoptotic DNA damage. In contrast, superoxide dismutase failed to inhibit Hcy-induced DNA damage. Taken together, these results demonstrate that Hcy exerted its genotoxic effects on HL-60 cells through an apoptotic pathway, which is mediated by the activation of caspase 3 activity induced by an increase in intracellular hydrogen peroxide.
Huang
R F
RF
Department of Nutrition and Food Sciences, Fu-Jen University, Hsin-Chuang, Taiwan, ROC. rweifen@mails.fju.edu.tw
Huang
S M
SM
Lin
B S
BS
Wei
J S
JS
Liu
T Z
TZ
D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000096:biosynthesis*; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000032:analysis; D004249:DNA Damage / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D004587:Electrophoresis, Agar Gel; D004789:Enzyme Activation; D005434:Flow Cytometry; D018922:HL-60 Cells / Q000187:drug effects* / Q000201:enzymology* / Q000473:pathology; D006710:Homocysteine / Q000031:analogs & derivatives* / Q000633:toxicity*; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D009152:Mutagenicity Tests; D009153:Mutagens / Q000633:toxicity*; D017382:Reactive Oxygen Species / Q000378:metabolism
D009153:Mutagens; D017382:Reactive Oxygen Species; D006710:Homocysteine; D004247:DNA; D006861:Hydrogen Peroxide; C007957:homocysteine thiolactone; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases