Purpose: The present study was conducted to re-evaluate the effect of low-level 1800 MHz RF signals (up to public exposure level for local exposure) on RAS/MAPK activation in live cells. Material and methods: Using molecular probes based on the Bioluminescence Resonance Energy Transfer technique (BRET), we assessed the effect of Continuous wave (CW) and Global System for Mobile (GSM)-modulated 1800 MHz signals (up to 2 W/kg) on ERK and RAS kinases' activity in live HuH7 cells. Results: We found that radiofrequency field (RF) exposure for 24h altered neither basal level of RAS and ERK activation nor the potency of phorbol-12-myristate-13-acetate (PMA) to activate RAS and ERK kinases, whatever the Specific Absorption Rate (SAR) or signal used. However, we found that exposure to GSM-modulated 1800 MHz signals at 2 W/kg decreased the PMA maximal efficacy to activate both RAS and ERK kinases' activity. Exposure with CW 1800 MHz signal at 2 W/kg only decreased maximal efficacy of PMA to activate ERK but not RAS. No effects of RF exposure at 0.5 W/kg was observed on maximal efficacy of PMA to activate either RAS or ERK whatever the signal used. Conclusion:Our results indicate that RF exposure decreases the efficiency of the cascade of events, which, from the binding of PMA to its receptor(s), leads to the activation of RAS and ERK kinases. This effect of RF exposure is reminiscent of RF-induced adaptive response.

In this study, a mode-stirred reverberation chamber (RC) was designed and proposed for the first time as a cell culture incubator for in vitro electromagnetic waves exposure of adherent cells in tissue culture plates. Typical cell incubators require specific conditions such as temperature of 37°C and humidity rate of 95 % which are challenging conditions for a RC. The chamber was characterized as an RC through an innovative experimental methodology based on the measurements of the S 11 parameter of the emitting antenna. The proposed RC is adapted for in vitro bioelectromagnetic experiments for simultaneous exposure of up to ten tissue culture plates under highly homogeneous exposure conditions at 3.5 GHz, i.e., the mid-frequency band of the 5G telecommunication networks. Results showed that the specific absorption rate (SAR) in the exposed samples extracted from temperature measurements was similar (an acceptable maximum variation lower than 30% was observed) in reason of the homogeneity and the uniformity of the field within the chamber. Specifically, measured SAR values were around 1.5 and 1 W/kg per 1 W of incident power, in 6-well or 96well tissue culture plates used for biological exposure, respectively.

It remains controversial whether exposure to environmental radiofrequency signals (RF) impacts cell status or response to cellular stress such as apoptosis or autophagy. We used two label-free techniques, cellular impedancemetry and Digital Holographic Microscopy (DHM), to assess the overall cellular response during RF exposure alone, or during co-exposure to RF and chemical treatments known to induce either apoptosis or autophagy. Two human cell lines (SH-SY5Y and HCT116) and two cultures of primary rat cortex cells (astrocytes and co-culture of neurons and glial cells) were exposed to RF using an 1800 MHz carrier wave modulated with various environmental signals (GSM: Global System for Mobile Communications, 2G signal), UMTS (Universal Mobile Telecommunications System, 3G signal), LTE (Long-Term Evolution, 4G signal, and Wi-Fi) or unmodulated RF (continuous wave, CW). The specific absorption rates (S.A.R.) used were 1.5 and 6 W/kg during DHM experiments and ranged from 5 to 24 W/kg during the recording of cellular impedance. Cells were continuously exposed for three to five consecutive days while the temporal phenotypic signature of cells behavior was recorded at constant temperature. Statistical analysis of the results does not indicate that RF-EMF exposure impacted the global behavior of healthy, apoptotic, or autophagic cells, even at S.A.R. levels higher than the guidelines, provided that the temperature was kept constant.