A Melanoma Cell Line Exposed To Extremely Low Frequency Magnetic Fields: Assessment Of Proliferation

ELIÇABE, JAVIER; LACOSTE, MARÍA GABRIELA; MAKINISTIAN, LEONARDO; GUERRA, MARÍA FLORENCIA; ANZULOVICH, ANA CECILIA

San Luis

Congreso; XXXVII Reunión Científica Anual de la Sociedad de Biología de Cuyo; 2019

In 2017 Kiessling et al. [BMC Biol. 2017;15(1):13]found that the clock genes are suppressed in B16 cells and tumours. Nevertheless, they were able to induce circadian rhythmicity with dexamethasone, forskoline and thermic shock. In consequence, B16 proliferation in vitro and tumour growth in vivo slowed down. Based on the literature, we pose that an alternative treatment for ?resynchronization? of circadian cycles in tumours could be the exposure to extremely low frequency electromagnetic fields (EL-EMF). Specifically, we hypothesize that: ?A treatment of 1 h with ELF-MF of 50 Hz and 0.1 mT decreases the proliferation and modifies the temporal expression of BMAL1 in B16 cells?. To test our hypothesis, we built two identical systems of coils of cubic geometry. Each one consisted of a triaxial system of 3 pairs of coils in an orthogonal arrange. We raised two types of experiments. In the first one, cells were seeded in 96-well microplates (one ?control? and one ?exposed?) and cell viability was measured by the MTT assay at T = 72 h (beginning of exposure was considered time zero, T = 0 h). A negative control (or sham-exposure) was first conducted and then a 50 Hz 100 µT AC field was tested under two different DC field configurations. No significant differences were found between both plates in any of the three experiments. In the second type of experiment, BMAL1 expression will be measured. For this, we will use 24 35mm dishes: 12 control and 12 exposed. The exposures will be identical to perform in viability assay. RNA will extract every 4 hours during 48 hs in a row, with the first extraction at T = 24 h after T0. In conclusion, the combination used of magnetic fields and duration of exposure did not affect the viability in B-16 cell line. After an intense study of literature, we conclude that the effects on cells depends on the ELF-MF intensity, duration of exposure and the cell line. In addition, study of BMAL1 expression will allow us to have an overview about the interaction between ELF-MF and circadian cycles.