| Keywords [eng] |
semaglutide, metformin, melanoma, SK-MEL-28, oxidative stress, reactive oxygen species, cell viability, cell migration, semaglutidas, metforminas, melanoma, SK-MEL-28, oksidacinis stresas, aktyviosios deguonies formos, ląstelių gyvybingumas, ląstelių migracija |
| Abstract [eng] |
Aim. To evaluate the effects of semaglutide combined with different concentrations of metformin on melanoma cell viability, oxidative stress and migration in vitro, and to compare these effects with those of semaglutide and metformin separately. Objectives. 1) To evaluate the effects of semaglutide, metformin and their combinations on melanoma cell viability in vitro; 2) To evaluate the effects of semaglutide, metformin and their combinations on oxidative stress in melanoma cells in vitro; 3) To evaluate the effects of semaglutide, metformin and their combinations on melanoma cell migration in vitro; 4) To compare the effects of semaglutide-metformin combinations with those of semaglutide and metformin separately on melanoma cell viability, oxidative stress and migration in vitro. Methods. Human melanoma SK-MEL-28 cells were treated with semaglutide (0.6 µM), metformin (2, 5, 10 mM) and combinations of semaglutide (0.6 µM) with metformin (2, 5, 10 mM). Cell viability was evaluated using the CCK-8 assay, oxidative stress by fluorometric measurement of intracellular reactive oxygen species (ROS) and migration using the wound healing assay. Statistical analysis was performed using one-way ANOVA with Dunnett’s or Tukey’s post hoc tests, or the Kruskal-Wallis test; p < 0.05 was considered statistically significant. Results. At 24 h, melanoma cell viability increased relative to control (100%) with metformin 5 mM (128.63 ± 0.76%, p = 0.021), semaglutide (0.6 µM) combined with metformin 2 mM (131.03 ± 6.83%, p = 0.012) or 5 mM (129.47 ± 14.03%, p = 0.007), whereas at 48 h, only metformin 5 mM showed a significant increase (140.20 ± 5.78%, p = 0.031). Intracellular ROS increased relative to control (100%) at 24 h with semaglutide (0.6 µM) (136.61 ± 7.16%, p = 0.019), metformin 5 mM (210.71 ± 1.68%, p < 0.001), metformin 10 mM (177.98 ± 18.52%, p < 0.001) and all combinations: 2 mM (141.07 ± 3.37%, p = 0.010), 5 mM (157.14 ± 11.79%, p = 0.001) and 10 mM (160.12 ± 3.37%, p = 0.001); at 48 h, only semaglutide (0.6 µM) showed a significant increase (149.84 ± 14.81%, p = 0.039). No significant differences in migration were observed. Combinations did not differ from semaglutide alone or metformin alone in their effects on viability or migration. For intracellular ROS levels, semaglutide (0.6 µM) combined with metformin 5 mM showed significantly lower ROS levels than metformin 5 mM alone at 24 h (−53.57%, p = 0.009), while at 48 h, all combinations showed significantly lower ROS levels than semaglutide (0.6 µM) alone (combination with 2 mM: −60.16%, p = 0.045; 5 mM: −70.63%, p = 0.020; 10 mM: −76.25%, p = 0.013). Conclusions. 1) Metformin 5 mM and semaglutide (0.6 µM) combined with metformin 2 mM or 5 mM all increased melanoma cell viability by approximately 30% at 24 h; however, only metformin 5 mM maintained this effect at 48 h, reaching an increase of approximately 40%. Semaglutide (0.6 µM) alone did not significantly affect cell viability. 2) Semaglutide (0.6 µM), metformin 5 mM and 10 mM, and all semaglutide-metformin combinations significantly increased intracellular reactive oxygen species (ROS) levels at 24 h. Metformin 5 mM showed the strongest increase in ROS, approximately 111%, while the combinations increased ROS by approximately 41-60% in a dose-dependent manner. At 48 h, only semaglutide (0.6 µM) maintained a significant increase in ROS by approximately 50%. 3) Semaglutide, metformin and their combinations did not significantly affect melanoma cell migration. 4) Semaglutide (0.6 µM) combined with metformin 5 mM showed significantly lower reactive oxygen species (ROS) levels than metformin 5 mM alone at 24 h. At 48 h, all semaglutide-metformin combinations showed significantly lower ROS levels than semaglutide (0.6 µM) alone. No significant differences between semaglutide-metformin combinations and semaglutide alone or metformin alone were observed in cell viability and migration. |
