Extremely low frequency electromagnetic fields affect proliferation and mitochondrial activity of human cancer cell lines
Published in: Int J Radiat Biol 2015; 91 (12): 964-972
Aim of study (acc. to author)
exposure duration and each treatment, a separate control group was used (remark EMF-Portal: no details were given on procedure for controls). Endpoint
continuous for 3, 6 or 7 days
cell culture dishes between coils in a box inside an incubator; the box shielded the cells and exposure apparatus from the background magnetic field and was made of stainless steel and the shield was made of a 2 mm thick inner layer of mu-metal and an outer layer made of a special aluminium-free alloy
setup was composed of two independent couples of coaxial
coils made of 200 loops (formed of copper wire, 0.3 mm in diameter) in the shielding box, each loop being 2.5 cm in length (with a resulting loop density of 8000 loops per meter) and wound into a tightly packed plastic frame; the frame had a cylindrical shape with an outer radius of 8 cm and the distance between the two coaxial coil couples was 8 cm; the cell culture dish was placed in the central part between the coils; the outer coils were supplied with a DC current and provided a static magnetic field of 45 µT to simulate the geomagnetic field; the inner coils (7.5 cm radius) were connected to an AC current generator ( signal with a duty cycle of 50%); inside the box, the temperature and humidity were homogenous to the rest of the incubator
magnetic field inside the shielding box measured around 1-2 µT
molecular biosynthesis: protein expression of phosphorylated ERK, ERK, p53, ErbB2/c-NEU ( marker for proliferation, especially in breast cancer cells), cytochrome C ( marker for mitochondrial permeability), VDAC ( mitochondrial pore protein) and PARP in whole cells, mitochondria and nuclei after 3 and 7 days of exposure in SKBR3 cells ( Western blot); gene expression of cytochrome c oxidase subunits (COX) II and IV after 3 and 7 days of exposure in SKBR3 and HT20 cells ( real-time RT-PCR)
cell function: mitochondrial activity in SKBR3 and HT29 cells: mitochondrial membrane potential after 3 and 7 days of exposure and after 6 days of exposure + 1 days without exposure and 3 days of exposure + 4 days without exposure (JC-1 stain, flow cytometry), amount of ATP in mitochondria after 7 days of exposure ( bioluminescence assay); glycolysis in cytosol ( exposure duration not specified; lactate dehydrogenase enzyme activity, spectrophotometry)
cell viability/ cell division/ proliferation: cell proliferation after 7 days of exposure in all cell lines ( crystal violet stain, spectrophotometry); apoptosis ( propidium iodide stain, fluorophotometry; sub-G0 peak in cell cycle distribution indicative of DNA fragmentation, flow cytometry) Main outcome of study (acc. to author)
Study funded by
European Union (EU)/European Commission
Ministero dell' Università e della Ricerca (MIUR (formerly MURST (Ministero dell' Università e della Ricerca Scientifica e Tecnologica); Ministry of University and Research), Italy
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