Potassium-channel activation in response to low doses of gamma-irradiation involves reactive oxygen intermediates in nonexcitatory cells.
Active oxygen species are generated during pathophysiologic conditions such as inflammation and ionizing radiation exposure. We tested the hypothesis that an early cellular event in response to these species involves regulation of ion channels. We exposed cells to gamma-irradiation or treated them with hydrogen peroxide, xanthine/xanthine oxidase, or [3H]thymidine and then monitored channel activity by the technique of whole-cell voltage clamping. Recordings showed that both normal and tumor cells exhibit an increase in K+ currents after treatment with radiation, H2O2, and xanthine/xanthine oxidase but not with high specific activity [3H]thymidine, suggesting that the signal for K+ channel activation originates at the cell membrane. A single noncytotoxic dose of 10 cGy induced measurable levels of K+ currents, suggesting that the induction of currents regulates biochemical changes in response to stress. To test whether channel activity is sensitive to active oxygen species, we pretreated cells with N-acetyl-L-cysteine (NAC) to increase cellular pools of free radical scavengers before radiation. In NAC-pretreated cells, K+ channel activation by gamma-irradiation was abolished. It has previously been shown that protein kinase C (PKC) is activated by ionizing radiation and can regulate K+ channels in some cells. However, the effect of radiation on induction of K+ channel activity was independent of PKC, since cells chronically exposed to phorbol esters still produced K+ currents after radiation. These results suggest that an early cellular response to oxidative stress is the activation of K+ channels.
Kuo
S S
SS
Department of Radiation Oncology, Stanford University School of Medicine, CA 94305.
Saad
A H
AH
Koong
A C
AC
Hahn
G M
GM
Giaccia
A J
AJ
D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D004307:Dose-Response Relationship, Radiation; D004553:Electric Conductivity; D016166:Free Radical Scavengers; D005609:Free Radicals; D005720:Gamma Rays; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008839:Microelectrodes; D010100:Oxygen / Q000378:metabolism*; D011188:Potassium / Q000378:metabolism*; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism / Q000528:radiation effects*; D011493:Protein Kinase C / Q000378:metabolism
D016166:Free Radical Scavengers; D005609:Free Radicals; D015221:Potassium Channels; D006861:Hydrogen Peroxide; D011493:Protein Kinase C; D011188:Potassium; D010100:Oxygen; D000111:Acetylcysteine