Decreasing of viability in H2O2treated of ITPA down-regulated human umbilical vein endothelial cells

Human inosine triphosphatase (ITPase), encoded by the ITPA gene is required for high-fidelity DNA and RNA replication. ITPA has been identified as a key gene involved in the removal deaminated nucleotides from the cellular nucleotide pool, maintains the stability of the genome. Defects in ITPA can result in inosine triphosphatase deficiency and accumulation of modified or damaged bases in genomic DNA or cellular RNAs that is a major cause of altered genetic information and mutagenesis.Oxidative deamination is a common chemical modification that damages DNA and RNA molecules. Free radicals of oxygen and nitrogen produced by oxidative stress in cells contribute to cell dysfunction via the apoptotic induction of endothelial cells (ECs). This study was focused on investigating the survival of stable ITPA down-regulated HUVEC compared to normal HUVEC in the presence of H2O2. To evaluate the cell viability assay, we used the 3-(4, 5-dimethylthiazol-2-yl) -2, 5- diphenyltetrazolium bromide (MTT) method. Briefly, 1x104 cells were incubated in a 96-well plate in the presence of various concentrations of H2O2 for 12-24 hours to determine the effect on endothelial cell proliferation. H2O2 decreases proliferative activity in ITPA down-regulated compared to normal HUVEC cells. The proliferation of treated cells was significantly lower than in the control wells (p<0.05).

Keywords:

Inosine triphosphatase (ITPA) , H2O2 , Endothelial cells