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8-Hydroxydeoxyguanosine in DNA from TPA-Stimulated Human Granulocytes
Authors:H C Birnboim  Lindsay Maidt  Tina Raynor  R A Floyd
Institution:  a Ottawa Regional Cancer Centre and University of Ottawa, Ottawa, Ontario, Canada b Free Radical Biology & Aging Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA
Abstract:8-Hydroxydeoxyguanosine (8-OHdG) is now widely used as a sensitive marker of oxidative damage to DNA. When human granulocytes are stimulated with TPA, they release a large quantity of reactive oxygen species (superoxide, hydrogen peroxide) which might be expected to generate hydroxyl radicals (OH-) which in turn could produce 8-OHdG in the DNA. There had been considerable debate as to whether OH -is detectable in stimulated granulocytes; most workers now agree that none can be detected, unless exogenous iron is added. An earlier report had described that 8-OHdG (a marker of OH -) was increased in the DNA of TPA-stimulated, compared to control, granulocytes. We have repeated this experiment and have been unable to reproduce this Finding. We conclude that the amount of 8-OHdG produced in the DNA of TPA-stimulated human ganulocytes is indistinguishable from that seen in control (unstimulated) cells (less than one 8- OHdG/105 dG).
Keywords:Granulocytes  hydroxyl radical  8-hydroxydeoxyguanosine  DNA strand breaks
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