Objective: The aim of the present study was to observe the effects of 1.8 GHz radiofrequency (RF) radiation on the protein expression of human lens epithelial cells (hLECs) in vitro.
Methods: The hLECs were exposed and sham-exposed to 1.8 GHz RF radiation (specific absorption rate (SAR) of 4 W/kg) for 2 h. After exposure, the proteins extracted from LECs were loaded on the Ettan MDLC system connected to the LTQ-Orbitrap MS for screening the candidate protein biomarkers induced by RF. The quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the levels of messenger RNA of candidate biomarkers. After the hLECs were exposed to 1.8 GHz RF (SAR of 2, 3 and 4 W/kg) for 2 h, the Western blot assay was utilized to measure the expression levels of the above-screened candidate protein biomarkers.
Results: The results of shotgun proteomic analysis indicated that there were eight proteins with differential expression between exposure and sham exposure groups. The results of qRT-PCR showed that there were three genes with expressional differences (valosin containing protein (VCP), ubiquitin specific peptidase 35 (USP35) and signal recognition particle 68 kDa (SRP68)) between exposure and sham exposure groups. The results of Western blot assay exhibited that the expressional levels of VCP and USP35 proteins significantly increased and the expressional level of protein SRP68 significantly decreased in hLECs exposed to 1.8 GHz RF radiation (SAR of 3 and 4 W/kg) for 2 h when compared with the corresponding sham groups (p < 0.05).
Conclusion: The shotgun proteomics technique can be applied to screen the proteins with differential expression between hLECs exposed to 1.8 GHz RF and hLECs sham-exposed to 1.8 GHz RF, and three protein biomarkers associated with RF radiation were validated by Western blot assay.
Keywords: Radiofrequency radiation; human lens epithelial cells; shotgun proteomics approach.