Combined effects of 872 MHz radiofrequency radiation and ferrous chloride on reactive oxygen species production and DNA damage in human SH‐SY5Y neuroblastoma cells

The aim of the present study was to investigate possible cooperative effects of radiofrequency (RF) radiation and ferrous chloride (FeCl(2)) on reactive oxygen species (ROS) production and DNA damage. In order to test intracellular ROS production as a possible underlying mechanism of DNA damage, we applied the fluorescent probe DCFH-DA. Integrity of DNA was quantified by alkaline comet assay. The exposures to 872 MHz RF radiation were conducted at a specific absorption rate (SAR) of 5 W/kg using continuous waves (CW) or a modulated signal similar to that used in Global System for Mobile Communications (GSM) phones. Four groups were included: (1) Sham exposure (control), (2) RF radiation, (3) Chemical treatment, (4) Chemical treatment, and RF radiation. In the ROS production experiments, human neuroblastoma (SH-SY5Y) cells were exposed to RF radiation and 10 microg/ml FeCl(2) for 1 h. In the comet assay experiments, the exposure time was 3 h and an additional chemical (0.015% diethyl maleate) was used to make DNA damage level observable. The chemical treatments resulted in statistically significant responses, but no effects from either CW or modulated RF radiation were observed on ROS production, DNA damage or cell viability.

[1]  O. Blokhina,et al.  Antioxidants, oxidative damage and oxygen deprivation stress: a review. , 2003, Annals of botany.

[2]  M. Simkó,et al.  Free Radical Release and HSP70 Expression in Two Human Immune-Relevant Cell Lines after Exposure to 1800 MHz Radiofrequency Radiation , 2006, Radiation research.

[3]  Ji-liang He,et al.  Study of low-intensity 2450-MHz microwave exposure enhancing the genotoxic effects of mitomycin C using micronucleus test and comet assay in vitro. , 2002, Biomedical and environmental sciences : BES.

[4]  C Bartsch,et al.  Chronic Exposure to a GSM-like Signal (Mobile Phone) Does Not Stimulate the Development of DMBA-Induced Mammary Tumors in Rats: Results of Three Consecutive Studies , 2002, Radiation research.

[5]  L. Verschaeve,et al.  954 MHz microwaves enhance the mutagenic properties of mitomycin C , 1996, Environmental and molecular mutagenesis.

[6]  Heinrich Ernst,et al.  No Effects of Radiofrequency Radiation on 3-Chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone-Induced Tumorigenesis in Female Wistar Rats , 2006, Radiation research.

[7]  R. Crebelli,et al.  Processing of DNA damage induced by hydrogen peroxide and methyl methanesulfonate in human lymphocytes: analysis by alkaline single cell gel electrophoresis and cytogenetic methods. , 1999, Mutagenesis.

[8]  A. Sannino,et al.  Human Fibroblasts and 900 MHz Radiofrequency Radiation: Evaluation of DNA Damage after Exposure and Co-exposure to 3-Chloro-4-(dichloromethyl)-5-Hydroxy-2(5h)-furanone (MX) , 2009, Radiation research.

[9]  P. Heikkinen,et al.  Micronucleus frequency in erythrocytes of mice after long-term exposure to radiofrequency radiation , 2007, International journal of radiation biology.

[10]  S. Loft,et al.  Menadione-induced DNA fragmentation without 8-oxo-2'-deoxyguanosine formation in isolated rat hepatocytes. , 1995, Biochemical pharmacology.

[11]  L. Verschaeve,et al.  Genetic effects of radiofrequency radiation (RFR). , 2005, Toxicology and applied pharmacology.

[12]  Lu Deqiang,et al.  Evaluating the combinative effects on human lymphocyte DNA damage induced by ultraviolet ray C plus 1.8 GHz microwaves using comet assay in vitro. , 2007, Toxicology.

[13]  P. Heikkinen,et al.  Investigation of Co-genotoxic Effects of Radiofrequency Electromagnetic Fields In Vivo , 2006, Radiation research.

[14]  Marek Zmyślony,et al.  Acute exposure to 930 MHz CW electromagnetic radiation in vitro affects reactive oxygen species level in rat lymphocytes treated by iron ions , 2004, Bioelectromagnetics.

[15]  D. McRee,et al.  Genotoxicity of radiofrequency signals. I. Investigation of DNA damage and micronuclei induction in cultured human blood cells , 2002, Bioelectromagnetics.

[16]  J. Juutilainen,et al.  Enhancement of chemically induced reactive oxygen species production and DNA damage in human SH-SY5Y neuroblastoma cells by 872 MHz radiofrequency radiation. , 2009, Mutation research.

[17]  K. Savolainen,et al.  Modification of glutamate-induced oxidative stress by lead: the role of extracellular calcium. , 1998, Free radical biology & medicine.

[18]  Jukka Juutilainen,et al.  Apoptosis induced by ultraviolet radiation is enhanced by amplitude modulated radiofrequency radiation in mutant yeast cells , 2004, Bioelectromagnetics.

[19]  E. Moros,et al.  Evaluation of Parameters of Oxidative Stress after In Vitro Exposure to FMCW- and CDMA-Modulated Radiofrequency Radiation Fields , 2004, Radiation research.

[20]  M Zmyślony,et al.  DNA damage in rat lymphocytes treated in vitro with iron cations and exposed to 7 mT magnetic fields (static or 50 Hz). , 2000, Mutation research.

[21]  Juan Ye,et al.  Electromagnetic noise inhibits radiofrequency radiation-induced DNA damage and reactive oxygen species increase in human lens epithelial cells , 2008, Molecular vision.

[22]  Ji-liang He,et al.  Studying the synergistic damage effects induced by 1.8 GHz radiofrequency field radiation (RFR) with four chemical mutagens on human lymphocyte DNA using comet assay in vitro. , 2005, Mutation research.

[23]  Guglielmo d'Ambrosio,et al.  Formation of Reactive Oxygen Species in L929 Cells after Exposure to 900 MHz RF Radiation with and without Co-exposure to 3-Chloro-4-(dichloromethyl)-5-hydroxy-2(5H)-furanone , 2007, Radiation research.

[24]  M. Martínez‐Búrdalo,et al.  Comparison of FDTD-calculated specific absorption rate in adults and children when using a mobile phone at 900 and 1800 MHz. , 2004, Physics in medicine and biology.

[25]  P. Heikkinen,et al.  Effects of mobile phone radiation on UV-induced skin tumourigenesis in ornithine decarboxylase transgenic and non-transgenic mice , 2003, International journal of radiation biology.

[26]  Vijayalaxmi,et al.  Genetic Damage in Mammalian Somatic Cells Exposed to Radiofrequency Radiation: A Meta-analysis of Data from 63 Publications (1990–2005) , 2008, Radiation research.

[27]  N. Kuster,et al.  ROS release and Hsp70 expression after exposure to 1,800 MHz radiofrequency electromagnetic fields in primary human monocytes and lymphocytes , 2006, Radiation and environmental biophysics.

[28]  Modest increase in temperature affects ODC activity in L929 cells: low-level radiofrequency radiation does not , 2006, Radiation and environmental biophysics.

[29]  A. Sannino,et al.  Reactive oxygen species formation is not enhanced by exposure to UMTS 1950 MHz radiation and co‐exposure to ferrous ions in Jurkat cells , 2009, Bioelectromagnetics.

[30]  Jukka Juutilainen,et al.  Pre-exposure to 50 Hz magnetic fields modifies menadione-induced DNA damage response in murine L929 cells , 2008, International journal of radiation biology.

[31]  Donald K Martin,et al.  An in vitro study of the effects of exposure to a GSM signal in two human cell lines: Monocytic U937 and neuroblastoma SK‐N‐SH , 2006, Cell biology international.

[32]  B. Veyret,et al.  No effect of short-term exposure to GSM-modulated low-power microwaves on benzo(a)pyrene-induced tumours in rat. , 1999, International journal of radiation biology.

[33]  P. Heikkinen,et al.  Effects of Mobile Phone Radiation on X-Ray-Induced Tumorigenesis in Mice , 2001, Radiation research.

[34]  M. Meltz,et al.  Radiofrequency (microwave) radiation exposure of mammalian cells during UV-induced DNA repair synthesis. , 1987, Radiation research.

[35]  M. Meltz,et al.  Influence of radiofrequency radiation on chromosome aberrations in CHO cells and its interaction with DNA-damaging agents. , 1990, Radiation research.

[36]  M Simkó,et al.  Hsp70 expression and free radical release after exposure to non-thermal radio-frequency electromagnetic fields and ultrafine particles in human Mono Mac 6 cells. , 2006, Toxicology letters.

[37]  Y. Ahuja,et al.  Combinative Exposure Effect of Radio Frequency Signals from CDMA Mobile Phones and Aphidicolin on DNA Integrity , 2008, Electromagnetic biology and medicine.

[38]  L. Verschaeve,et al.  Cytogenetic effects of 900 MHz (GSM) microwaves on human lymphocytes. , 2001, Bioelectromagnetics.

[39]  M. Meltz,et al.  Absence of mutagenic interaction between microwaves and mitomycin C in mammalian cells , 1989, Environmental and molecular mutagenesis.

[40]  M. Geffard,et al.  Effects of GSM-900 Microwaves on DMBA-Induced Mammary Gland Tumors in Female Sprague-Dawley Rats , 2003, Radiation research.

[41]  Niels Kuster,et al.  Effects of 900 MHz GSM Wireless Communication Signals on DMBA-Induced Mammary Tumors in Rats , 2006, Radiation research.

[42]  Georg Neubauer,et al.  Study on potential effects of "902-MHz GSM-type Wireless Communication Signals" on DMBA-induced mammary tumours in Sprague-Dawley rats. , 2008, Mutation research.