The effect of ouabain on mitochondrial DNA damage in HepG2 cell lines.
Our purpose in this study is to analyze mitochondrial DNA (mtDNA) lesion frequencies and mtDNA(4977) deletion in HepG2 cells to examine the effects of ouabain on mtDNA. HepG2 cells were treated with 0.75, 7.5, 75, and 750 nM of ouabain for 24 h in the presence and absence of 10 mM 2-deoxyglucose (2-DG). The frequency of mtDNA(4977) deletions and mitochondrial lesions were determined by real-time polymerase chain reaction. A ≥ 1.2-fold change or greater was considered significant. Ouabain doses of 750, 75, and 7.5 nM alone increased the frequency of mtDNA(4977) deletions 1.39, 1.92, and 1.44 times, respectively. The 750 and 75 nM ouabain doses combined with 2-DG increased the mtDNA(4977) deletion frequency 4.94 and 1.57 times, respectively. The 750 and 75 nM ouabain doses alone increased the mtDNA lesion frequency 2.5 and 1.5 times, respectively. The 750 nM ouabain dose combined with 2-DG increased the mtDNA lesion frequency 2.28 times. The 7.5 nM ouabain dose alone and combined with 2-DG decreased the mtDNA lesion frequency 0.67 and 0.45 times, respectively. Ouabain alone and when combined with 2-DG increases mtDNA lesion and mtDNA(4977) deletion frequencies. This supports the thesis that ouabain creates oxidative stress and induces DNA damage and apoptosis.
Aksaray State Hospital, Aksaray, Turkey.
Rukiye
R
Nar
Abdulkerim
A
Bedir
Hasan
H
Alacam
Veli
V
Kilinc
Bahattin
B
Avci
Osman
O
Salis
Sedat
S
Gulten
D002316:Cardiotonic Agents / Q000494:pharmacology*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004249:DNA Damage / Q000187:drug effects* / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics*; D003847:Deoxyglucose / Q000494:pharmacology; D056945:Hep G2 Cells; D006801:Humans; D008928:Mitochondria / Q000187:drug effects* / Q000235:genetics / Q000473:pathology; D010042:Ouabain / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects*; D016133:Polymerase Chain Reaction; D060888:Real-Time Polymerase Chain Reaction
D002316:Cardiotonic Agents; D004272:DNA, Mitochondrial; D010042:Ouabain; D004247:DNA; D003847:Deoxyglucose