Mechanism of thymidine incorporation into acid insoluble fraction via nucleoside transporters on oxidative stress DNA injury

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  • Tanaka Koh-ichi
    Division of Pharmacology, Department of Pharmacy, School of Pharmacy, Hyogo University of Health Sciences, Japan Department of Pharmacology, Hyogo College of Medicine, Japan Department of Applied Pharmacology, Kagoshima University Graduate School of Medical and Dental Sciences, Japan
  • Kitanaka Nobue
    Department of Pharmacology, Hyogo College of Medicine, Japan
  • Kitanaka Junichi
    Department of Pharmacology, Hyogo College of Medicine, Japan
  • Tomita Kazuo
    Division of Pharmacology, Department of Pharmacy, School of Pharmacy, Hyogo University of Health Sciences, Japan Department of Applied Pharmacology, Kagoshima University Graduate School of Medical and Dental Sciences, Japan
  • Tsukahara Takao
    Department of Applied Pharmacology, Kagoshima University Graduate School of Medical and Dental Sciences, Japan
  • Sato Tomoaki
    Department of Applied Pharmacology, Kagoshima University Graduate School of Medical and Dental Sciences, Japan
  • Takemura Motohiko
    Department of Pharmacology, Hyogo College of Medicine, Japan
  • Nishiyama Nobuyoshi
    Division of Pharmacology, Department of Pharmacy, School of Pharmacy, Hyogo University of Health Sciences, Japan

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<p>We have found that cultured astrocytes pretreated with N6, 2'-O-dibutyryladenosine 3',5'-cyclic monophosphate (DBcAMP), a permeable analogue of cAMP, but not astrocytes pretreated without DBcAMP and neurons, have the ability to incorporate thymidine into acid insoluble fraction via nucleoside transporters at an early time for repair on hydrogen peroxide (H2O2)-induced DNA injury.</p><p>We studied expression of equilibrative nucleoside transporter (ENT) and concentrative nucleoside transporter (CNT) and the relation between thymidine incorporation into intracellular spaces (membrane transport) and acid insoluble fraction (DNA repair) on cultured astrocytes pretreated with DBcAMP in the presence and absence of H2O2. We concerned that astrocytes express ENT1, ENT2, and CNT2, CNT3, but not CNT1, by western blot, immunocytochemistry and RT-PCR, and H2O2 caused decrease in membrane transport of thymidine from extracellular spaces to intracellular spaces and increase in incorporation of thymidine into acid insoluble fraction to cultured astrocytes pretreated with DBcAMP.</p><p>These finding indicate that cultured differentiated astrocytes could incorporate thymidine effectively into acid insoluble fraction for repair on H2O2-induced DNA injury, although the function of ENT2 and CNT3 might be impaired.</p>

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