Melanoma cell migration in response to red and near-infrared low-level light

Abstract

Cell migration plays an important role in tissue formation and cancer progression. In vitro scratch assay has been used for many years to study cell migration to mimic the migration of in vivo cells, and, thus, to evaluate cancer growth. Low-level red and near-infrared light (LLL) can increase normal cell migration. However, the impact of LLL on tumor cells remains unclear. In this work, we aimed to evaluate the effects of a single LLL dose on melanoma cell migration. B16F10 (murine melanoma) cells were cultivated in RPMI medium with 10% of fetal bovine serum until they reached 80% confluency. The cell line was seeded in a 6-well plate at a density of 2x10 5 cells/well in triplicate at two different moments. A wound scratch was performed to disrupt the confluent cell monolayer with a 10 μL pipette tip. Immediately after the injury, the cells were submitted to the LLL at two distinct wavelengths (660 and 780 nm) provided by a LED and a laser, respectively, delivering 3 different energies (1.3, 3.6, and 6 J) at an irradiance of 4.2 mW/cm2. The control group was not irradiated. Cells were photographed immediately and at 3, 12, 24, and 36 h after the scratch. The wound closure was measured using ImageJ software. To evaluate the overall migration, we calculated the areas under the curve for each group. Cells exposed to the red laser at 6 J migrated slower than control. In contrast, LLL at 780 nm promoted faster cell migration when irradiated with 3.6 J. These results suggest that low-level LEDs at 660 nm could prevent melanoma progression in higher energies. However, 780 nm should be avoided at middle energies.