Jornal Brasileiro de Pneumologia (Dec 2005)
Efeito do álcool perílico na expressão gênica de células de adenocarcinoma de pulmão humano Effect of perillyl alcohol on gene expression of human pulmonary adenocarcinoma cells
Abstract
OBJETIVO: Estudar a ação do álcool perílico na expressão gênica de células de adenocarcinoma de pulmão humano. MÉTODOS: Incubaram-se células de adenocarcinoma de pulmão com álcool perílico em diluições que variaram entre 0,03% e 0,0003% por 48 horas. Observaram-se as alterações na morfologia celular e quantificou-se a viabilidade celular pelo método do MTT (3-(4,5-dimetiltiazol-2-yl)-2,5 difeniltertrazolim brometo). Analisou-se a síntese de proteínas das amostras previamente marcadas radioativamente com 35S, através de eletroforese em gel de poliacrilamida. Determinou-se a expressão das proteínas p53 e p42/44 através do método de Western Blot. RESULTADOS: Após 48 horas de incubação, observaram-se alterações na morfologia celular para a diluição de 0,03% de álcool perílico, as quais foram pouco verificadas em diluições superiores a 0,003%. A inibição da viabilidade celular foi de 60,17% (p OBJECTIVE: To study the effect of perillyl alcohol on the gene expression of human pulmonary adenocarcinoma cells. METHODS: Pulmonary adenocarcinoma cells were incubated with perillyl alcohol in dilutions ranging from 0.03% to 0.0003% for 48 hours. Alterations were observed in the cell morphology, and cell viability was quantified using [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assays. Protein synthesis of samples previously targeted with S35 was analyzed using electrophoresis on a polyacrylamide gel. Expression of the proteins p53 and p44/42 was determined using the Western blot method. RESULTS: After 48 hours of incubation, greater nsumbers of morphological alterations were observed in cells treated with the 0.03% perillyl alcohol dilution than in those treated with perillyl alcohol diluted to 0.003% or further. Treatment with perillyl alcohol dilutions of 0.03%, 0.003% and 0.0003% inhibited cellular viability by 60.17% (p < 0.001), 15.62% (p < 0.001) and 11.53% (p < 0.05), respectively. The results show that 28-kDa, 42-kDa and 110-kDa proteins were induced. No statistically significant effect on p53 expression was observed. In comparison with the expression of alpha-tubulin, the 0.003% perillyl alcohol dilution induced an increase in p42 phosphorylation and a marked decrease in p44 phosphorylation. CONCLUSION: The results suggest that there are other, previously undescribed, metabolic pathways for perillyl alcohol effects in human pulmonary adenocarcinoma cells.
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