| Abstract [eng] |
Relevance of the research: Melanoma is a highly aggressive skin cancer with a strong metastatic potential, and its incidence continues to rise worldwide. Since oxidative stress plays an important role in melanoma progression, investigating the effects of alpha-lipoic acid and vitamin B1 in in vitro melanoma cell models may contribute to a better understanding of their impact on oxidative stress mechanisms. Aim of the study: To evaluate the effects of alpha-lipoic acid and vitamin B1 on oxidative stress-induced damage in in vitro melanoma cell models. Objectives of the study: 1. To evaluate the effect of alpha-lipoic acid on proliferation, reactive oxygen species levels, and migration in an oxidative stress-affected in vitro melanoma cell model. 2. To evaluate the effect of vitamin B1 on proliferation, reactive oxygen species levels, and migration in an oxidative stress-affected in vitro melanoma cell model. 3. To compare the effects of alpha-lipoic acid and vitamin B1 on proliferation, migration, and reactive oxygen species levels in an oxidative stress-affected in vitro melanoma cell model. Methods: SK-MEL-28 melanoma cell line was used and cultured under standard conditions in DMEM. Cell viability was determined using a Neubauer chamber and the trypan blue exclusion method. The effects of the tested compounds were evaluated using different concentrations of ALA (1 µM, 500 µM, 1 mM) and B1 (10 µM, 500 µM, 1 mM). Cell proliferation was analyzed using the CCK-8 assay. ROS levels were determined by measuring fluorescence intensity. Migration activity was assessed by analyzing cell area closure (%) every 12 h. Experiments were performed in three independent replicates. Statistical analysis was conducted using the ,,R Commander” software, applying ANOVA and Dunnett’s test to compare experimental groups with the control group. Statistical significance was considered at p ≤ 0.05. Graphical analysis was performed using ,,Microsoft Excel”. Results: After 48 h, ALR significantly reduced ROS levels in all tested groups compared to the control (p < 0,001), with reductions ranging from 52% to 54%. ALR also suppressed cell migration after 36 h by 63-84%, depending on concentration: 1 µM (p = 0,0028), 500 µM (p = 0,0024), and 1 mM (p = 0,0062). Following 48 h of incubation, migration inhibition remained evident, reaching 54.5-57% (p = 0,0179-0,0387). B1 10 µM increased proliferation by 30% (p = 0,0246), whereas 1 mM reduced it by 53% (p = 0,0009). After 72 h, proliferation inhibition reached 31.5% at B1 10 µM (p = 0,0014), 77,5% at B1 500 µM, and 82,5% at B1 1 mM (p < 0,0001). In addition, oxidative stress decreased by 70-82,5% after 48 h (p < 0,001), compared with the control group. Vitamin B1 also inhibited cell migration by 63-94% after 36 h and by 63,5-92% after 48 h (p < 0,001), depending on concentration. Conclusions: 1. Alpha-lipoic acid reduced oxidative stress by half compared to the control group and inhibited cell migration by up to 84% in vitro. 2. At lower concentrations, vitamin B1 increased proliferation by nearly one-third, whereas higher concentrations inhibited proliferation by up to 53%. With prolonged incubation time, proliferation was inhibited by up to 82.5%. In addition, vitamin B1 reduced oxidative stress by up to 82.5% and, at higher concentrations, inhibited migration by up to 94% in vitro. 3. Comparison of the effects of the investigated compounds on oxidative stress-induced melanoma cells demonstrated that alpha-lipoic acid and vitamin B1 exerted different effects on cell proliferation, ROS levels, and migration in vitro. |
