Measurements of Alkali-Labile DNA Damage and Protein–DNA Crosslinks after 2450 MHz Microwave and Low-Dose Gamma IrradiationIn Vitro

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Abstract
Lagroye, I., Hook, G. J., Wettring, B. A., Baty, J. D., Moros, E. G., Straube, W. L. and Roti Roti, J. L. Measurements of Alkali-Labile DNA Damage and Protein–DNA Crosslinks after 2450 MHz Microwave and Low-Dose Gamma Irradiation In Vitro. Radiat. Res. 161, 201–214 (2004).In vitro experiments were performed to determine whether 2450 MHz microwave radiation induces alkali-labile DNA damage and/or DNA–protein or DNA–DNA crosslinks in C3H 10T½ cells. After a 2-h exposure to either 2450 MHz continuous-wave (CW) microwaves at an SAR of 1.9 W/kg or 1 mM cisplatinum (CDDP, a positive control for DNA crosslinks), C3H 10T½ cells were irradiated with 4 Gy of γ rays (137Cs). Immediately after γ irradiation, the single-cell gel electrophoresis assay was performed to detect DNA damage. For each exposure condition, one set of samples was treated with proteinase K (1 mg/ml) to remove any possible DNA–protein crosslinks. To measure DNA–protein crosslinks independent of DNA–DNA crosslinks, we quantified the proteins that were recovered with DNA after microwave exposure, using CDDP and γ irradiation, positive controls for DNA–protein crosslinks. Ionizing radiation (4 Gy) induced significant DNA damage. However, no DNA damage could be detected after exposure to 2450 MHz CW microwaves alone. The crosslinking agent CDDP significantly reduced both the comet length and the normalized comet moment in C3H 10T½ cells irradiated with 4 Gy γ rays. In contrast, 2450 MHz microwaves did not impede the DNA migration induced by γ rays. When control cells were treated with proteinase K, both parameters increased in the absence of any DNA damage. However, no additional effect of proteinase K was seen in samples exposed to 2450 MHz microwaves or in samples treated with the combination of microwaves and radiation. On the other hand, proteinase K treatment was ineffective in restoring any migration of the DNA in cells pretreated with CDDP and irradiated with γ rays. When DNA–protein crosslinks were specifically measured, we found no evidence for the induction of DNA–protein crosslinks or changes in amount of the protein associated with DNA by 2450 MHz CW microwave exposure. Thus 2-h exposures to 1.9 W/ kg of 2450 MHz CW microwaves did not induce measurable alkali-labile DNA damage or DNA–DNA or DNA–protein crosslinks.

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