CAROLINA GOUVEA DE SOUZA CONTATORI
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Resumo IPEN-doc 28607 Melanoma cell migration in response to red and near-infrared low-level light2021 - CONTATORI, CAROLINA G. de S.; PINTO, MAYARA S.; RIBEIRO, MARTHA S.INTRODUCTION 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. OBJECTIVES In this work, we aimed to evaluate the effects of a single LLL dose on melanoma cell migration. MATERIALS AND METHODS 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 cellmonolayerwith 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/cm 2 . 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. DISCUSSION AND RESULTS 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. CONCLUSION 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.Resumo IPEN-doc 26888 Effects of low-level laser irradiation on VEGF expression of melanoma cell lines2020 - CONTATORI, C.G. de S.; SILVA, C.R.; YOSHIMURA, T.M.; RIBEIRO, M.S.Impact of low-level laser irradiation on tumor cell lines remains controversial. Vascular endothelial growth factor (VEGF) is a key molecule to form new blood vessels, which contribute for cancer development and growth. The aim of this study was to evaluate the effects of different light fluences on human melanoma SKMEL 37 cells and murine melanoma B16F10 cells using a near infrared laser (λ= 780 nm) with output power of 40 mW delivering energies of 1.2, 3.6 and 6 J (fluences of 30, 90 and 150 J/cm2, respectively). The cell lines were irradiated 24 h after they were seeded in a 96-well plate at a density of 5x103 cells per well, in triplicate at three different days. Following irradiation, both cell line supernatants were stored in Eppendorf tubes at - 20°C until VEGF-A expression measurement. Specific ELISA kits were used according to cell line (murine or human). Samples and standard solutions were added in a 96-well plate antibody-coated and incuba ted over night at 4°C. Reagent dilution and set time followed fabricant instructions. The stop solution was added and the absorbance was read in a microplate reader at 450 nm. Results showed a non-statistically significant difference among treated and control groups for both cell lines. These findings indicate that irradiation with near infrared laser does not influence VEGF expression on melanoma cell lines regardless the fluence used and should be tested to prevent cancer growth in preclinical assays.