Oxidative damage to DNA induced by areca nut extract.
In Taiwan, people chew betel quid which contains tender areca nut with husk. In other countries, people prefer ripe and dried areca nut without husk. In this study, we compared the reactive oxygen species-induced oxidative DNA damage in isolated DNA and CHO-K1 cells between treatments with tender areca nut extract (ANE) and ripe ANE. Incubation of these two ANE preparations with isolated DNA generated 8-hydroxy-2'-deoxyguanosine (8-OH-dG) in an alkaline environment in a dose-dependent manner. Ripe ANE generated higher levels of 8-OH-dG compared to tender ANE. The addition of iron(II) (100 microM) resulted in 1.4- and 3.1-fold increases of 8-OH-dG when incubated with 1 mg/ml each of tender and ripe ANE. In testing the effect of ANE to cellular DNA, CHO-K1 cells were used for its documented sensitivity to reactive oxygen species. In CHO-K1 cells, ripe ANE was more cytotoxic than tender ANE following an 18-h incubation. The cytotoxicity to CHO-K1 cells was positively correlated with the formation of 8-OH-dG following tender (r=0.97) and ripe (r=0.91) ANE treatment. Addition of the iron chelating agent o-phenanthroline (10 and 20 microM) to cells prior to ri ANE exposure significantly increased (p<0.05) the survival of CHO-K1 cells. In addition, ripe ANE induced dichlorofluorescein-mediated fluorescence which indicated the formation of hydrogen peroxide in CHO-K1 cells. In conclusion, this study demonstrated that ANE-induced oxidative damage to isolated and cellular DNA which may result from the generation of hydrogen peroxide, and iron may serve as a catalyst in this process. Furthermore, ripe ANE generated higher oxidative DNA damage levels compared to tender ANE.
Liu
T Y
TY
Department of Medical Research, Veterans General Hospital, Taipei, Taiwan.
Chen
C L
CL
Chi
C W
CW
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D001114:Areca; D016466:CHO Cells; D016923:Cell Death / Q000187:drug effects; D006224:Cricetinae; D004249:DNA Damage; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D005290:Ferric Compounds / Q000494:pharmacology; D005399:Fishes; D005453:Fluorescence; D006861:Hydrogen Peroxide / Q000378:metabolism; D008297:Male; D009754:Nuts / Q000633:toxicity*; D018384:Oxidative Stress; D010618:Phenanthrolines / Q000494:pharmacology; D010936:Plant Extracts / Q000633:toxicity; D010946:Plants, Medicinal; D017382:Reactive Oxygen Species / Q000378:metabolism; D013094:Spermatozoa
D005290:Ferric Compounds; D010618:Phenanthrolines; D010936:Plant Extracts; D017382:Reactive Oxygen Species; D000080242:8-Hydroxy-2'-Deoxyguanosine; D006861:Hydrogen Peroxide; D003849:Deoxyguanosine; C025205:1,10-phenanthroline