Regulation of a novel cell differentiation-associated gene, JWA during oxidative damage in K562 and MCF-7 cells.
Oxidative stress, or the production of oxygen-centered free radicals, has been hypothesized as the major source of DNA damage that can lead to a variety of diseases including cancer. It is known that 8-hydroxy-deoxyguanosine (8-oxo-dG) is a useful biomarker of oxidative DNA damage. Our recent data showed that JWA, initially being cloned as a novel cell differentiation-associated gene, was also actively responsive to environmental stressors, such as heat-shock, oxidative stress and so on. In the present study, we have applied a modified comet assay and bacterial repair endonucleases system (endonuclease III and formamidopyrimidine glycosylase) to investigate if JWA is involved in hydrogen peroxide (H2O2)-induced DNA damage and repair in K562 and MCF-7 cells, and to demonstrate if the damage is associated with 8-oxo-dG. The results from the comet assay have shown that the average tail length and the percentage of the cells with DNA tails are greatly induced by H2O2 treatment and further significantly enhanced by the post-treatment of repair endonucleases. The H2O2-induced 8-oxo-dG formation in K562 and MCF-7 cells is dose-dependent. In addition, the data have clearly demonstrated that JWA gene expression is actively induced by H2O2 treatment in K562 and MCF-7 cells. The results suggest that JWA can be regulated by oxidative stress and is actively involved in the signal pathways of oxidative stress in the cells.
Zhu
Ting
T
Department of Molecular Cell Biology and Toxicology, Jiangsu Provincial Key Laboratories of Human Functional Genomics and of Applied Toxicology; School of Public Health, Nanjing Medical University, 140 Hanzhong Road, Nanjing 210029, China.
Chen
Rui
R
Li
Ai-Ping
AP
Liu
Jia
J
Liu
Qi-Zhan
QZ
Chang
Hebron C
HC
Zhou
Jian-Wei
JW
D000080242:8-Hydroxy-2'-Deoxyguanosine; D002454:Cell Differentiation / Q000235:genetics*; D045744:Cell Line, Tumor; D002470:Cell Survival; D020552:Comet Assay; D004249:DNA Damage; D004260:DNA Repair; D043382:DNA-Formamidopyrimidine Glycosylase / Q000378:metabolism; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D043224:Deoxyribonuclease (Pyrimidine Dimer) / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D029968:Escherichia coli Proteins / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins; D020014:K562 Cells; D026901:Membrane Transport Proteins; D016877:Oxidants / Q000494:pharmacology; D018384:Oxidative Stress; D015398:Signal Transduction / Q000502:physiology
C484566:ARL6IP5 protein, human; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; D026901:Membrane Transport Proteins; D016877:Oxidants; D000080242:8-Hydroxy-2'-Deoxyguanosine; D006861:Hydrogen Peroxide; D043224:Deoxyribonuclease (Pyrimidine Dimer); C106403:NTH protein, E coli; D043382:DNA-Formamidopyrimidine Glycosylase; D003849:Deoxyguanosine