Mechanism of radiation-induced diacylglycerol production in primary cultured rat hepatocytes.
Protein kinase C (PKC) is known to be a key enzyme in radiation-induced signal transduction pathways. We have previously demonstrated that gamma-irradiation induces PKC activation and translocation from cytosol to membranes as a consequence of membrane lipid peroxidation in cultured rat hepatocytes (Int. J. Radiat. Biol. 70, 473-480, 1996). The present study was undertaken to investigate production of diacylglycerol, an endogenous activator of PKC, following gamma-irradiation of hepatocytes. Diacylglycerol content increased 3 min after irradiation, then decreased at 15 min and increased again at 30 min, indicating a biphasic pattern. This result implies participation of diacylglycerol in the radiation-induced activation of PKC in hepatocytes. In order to clarify the mechanism of the initial process of radiation-induced diacylglycerol production, the effects of reactive oxygens were investigated. Treatment of cells with hydroxyl radical, a major oxygen radical produced by radiation, induced diacylglycerol production without any change in the content of phosphatidylcholine, showing a peak at 1 min after treatment. No change in the diacylglycerol content was observed at that time by hydrogen peroxide treatment. Furthermore, the diacylglycerol production by hydroxyl radical was inhibited by pretreatment with neomycin sulfate, a phosphatidylinositol-specific phospholipase C (PI-PLC) inhibitor. These results suggest that radiation exerts PI-PLC activation through hydroxyl radical generation, followed by diacylglycerol production and PKC activation.
Nakajima
T
T
Division of Biology and Oncology, National Institute of Radiological Sciences, Chiba-shi, Japan. otetsu@nirs.go.jp
Yukawa
O
O
D000818:Animals; D002478:Cells, Cultured; D004075:Diglycerides / Q000096:biosynthesis*; D004789:Enzyme Activation / Q000528:radiation effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005720:Gamma Rays; D017665:Hydroxyl Radical / Q000494:pharmacology; D008099:Liver / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects*; D009355:Neomycin / Q000494:pharmacology; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors
D004075:Diglycerides; D004791:Enzyme Inhibitors; D017665:Hydroxyl Radical; D011493:Protein Kinase C; D010738:Type C Phospholipases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D009355:Neomycin