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The effects of exposure of hepatic carcinoma cells to a 50 Hz magnetic field and a treatment with melatonin on cell viability, proliferation and differentiation should be investigated.
Cells were exposed with or without melatonin (10 nM or 1 µM). For each group, a respective sham exposure was conducted. Cells were investigated directly after exposure. Additionally, cells were treated with melatonin alone (0.01 nM - 1 µM), without exposure to the magnetic field or sham exposure. Cells of these groups were investigated 5 days after plating.
細胞生存力／細胞分裂/増殖: cell viability, proliferation and differentiation of carcinoma cells
60 mm petri dishes in two identical, magnetically shielded chambers (Co-NETIC metal) located in two identical CO2 incubators
two identical pairs of coils connected in series to produce a vertically polarized and homogeneous magnetic field were placed inside the shielded chambers; each coil was made of 1000 turns of enameled copper wire; petri dishes were placed in the center of the coils; the incubator maintained a 5% CO2 and 100% humidity atmosphere at 37°C
A sham exposure was conducted.
in each experimental run, only one of the two coil sets was energized for MF exposure and the other one was used for sham exposure; both incubators were used alternatively for MF-exposure or sham exposure, in a random sequence
細胞生存力／細胞分裂/増殖: cell viability (trypan blue exclusion, haemocytometer, microscopy), cell proliferation (bromodeoxyuridine incorporation and protein expression of PCNA (proliferating cell nuclear antigen - a proliferation marker), fluorescence microscopy); cell differentiation (protein expression of alpha-fetoprotein (a marker for poorly differentiated hepatocytes), Western blot; amount of basal albumin (marker for adult hepatocytes; immunocytochemistry, optical turbidity measurement (nephelometry))
The cell viability did not show any significant differences between any exposure or melatonin-treated group and the respective sham exposure or control groups. However, exposure to the magnetic field alone resulted ina significantly increased cell proliferation and a significantly decreased differentiation compared to the sham exposure groups. A treatment with a very low dose of melatonin (0.01 nM) resulted in a significantly increased cell proliferation as well, higher concentrations (0.2 nM - 1 µM) led to a significantly decreased proliferation and a significantly increased differentiation (10 nM and 1 µM) compared to the control group. In the co-exposure groups, melatonin (10 nM and 1 µM) led to a significant reduction or even a suppression of the proliferation-promotive effect of the magnetic field exposure. Regarding differentiation, however, only a small dose of melatonin (10 nM) reversed the decreased differentiation effect of the magnetic field exposure. Higher concentrations of melatonin in combination with the magnetic field (1 µM) did not show this antagonistic effect and led instead to the same decreased differentiating effects as found in the magnetic exposure alone. The authors conclude, that exposure to a 50 Hz magnetic field and a treatment with melatonin might have partially antagonistic effects on cell proliferation and differentiation in hepatic carcinoma cells. As a whole, the results suggest that weak extremely low frequency magnetic fields could influence cancer-related processes in vitro, and reinforce the hypothesis that these fields could interfere with the potentially anti-tumoral action of melatonin.