RESUMO
AIM: To analyze involvement of human beta-defensin-2 (hBD-2) in intracellular signaling in vitro. MATERIALS AND METHODS: A431cells were cultured in the presence of 1 microg/ml of recombinant hBD-2 and/or 10 ng/ml EGF. For evaluation of expression of mRNAs for p70S6 kinase, isoforms alpha and beta, RT-PCR analysis was applied. Expression and activity of p70S6K, phosphorylation of PDK1, ERK, JNK, p38 kinases and EGF receptor (EGFR) was evaluated using Western blot analysis. RESULTS: 30 min incubation of A431 cells with 1 mug/ml of hBD-2 didn't influence autophosphorylation level of EGFR, but resulted in activation of p70S6K, 12 h treatment - in prominently increased level of mRNA for alpha and beta-isoforms of p70S6 kinase, whilst 24 h treatment - in elevation of p70S6K synthesis on protein level. Up-stream kinase phosphorylating p70S6K, PDK1, is also phosporylated upon influence of exogenous hBD-2 in vitro. CONCLUSION: Our data point on the involvement of PDK1-p70S6K pathway in mediation of action of hBD-2 in A431 cells.
Assuntos
Neoplasias/enzimologia , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Quinases S6 Ribossômicas 70-kDa/metabolismo , beta-Defensinas/farmacologia , Proteínas Quinases Dependentes de 3-Fosfoinositídeo , Ativação Enzimática , Fator de Crescimento Epidérmico/farmacologia , Receptores ErbB/agonistas , Humanos , Fosforilação/efeitos dos fármacos , Transdução de Sinais , Células Tumorais CultivadasRESUMO
The normalising effect of low-intensive electromagnetic radiation on the viability of cell culture of L929 line, irradiated with gamma-quantums of 137Cs in a wide dose range was demonstrated.
Assuntos
Fibroblastos/patologia , Fibroblastos/efeitos da radiação , Animais , Divisão Celular/efeitos da radiação , Linhagem Celular , Sobrevivência Celular/efeitos da radiação , Raios gama , Camundongos , Doses de Radiação , Radiação IonizanteRESUMO
The dependence of some parameters of L-cells culture viability on different concentrations of heavy metals was studied. Considerable cytotoxic effect of low concentrations of nickel (0.025 mcg/ml) and lead (0.05 mcg/ml) was shown. Copper and chrome at concentrations of 0.25-0.5 mcg/ml promote cells proliferation between third and fifth days of cultivation. Nickel at concentration 0.025 mcg/ml and lead at all investigated concentrations synchronize cells division in culture. Increasing of giant polynucleas cells level in culture was characteristic for investigated metals. The maximum levels of this type cells were caused by the action of nickel, chrome and copper.
