RESUMO
In this report we determine the ability of ursolic acid (UA) to induce apoptosis and to modulate glucocorticoid receptor (GR) and Activator Protein-1 (AP-1) in MCF-7 cells. The UA-induced apoptosis (53 microM), the PARP cleavage, and the decrease in Bcl-2 protein (53 microM) support the notion that UA induces apoptosis through the intrinsic mitochondrial pathway. UA binds GR (relative binding affinity: 2.57) and translocates GR into nucleus, suggesting its potential as a GR modulator. UA had no effect on GRE- or TRE-driven gene expression. In summary, UA is a GR modulator and may be considered as a potential anticancer agent in breast cancer.
Assuntos
Adenocarcinoma/patologia , Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias da Mama/patologia , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Triterpenos/farmacologia , Transporte Ativo do Núcleo Celular , Adenocarcinoma/genética , Ligação Competitiva , Neoplasias da Mama/genética , Linhagem Celular Tumoral/efeitos dos fármacos , Linhagem Celular Tumoral/metabolismo , Linhagem Celular Tumoral/patologia , Dexametasona/farmacologia , Regulação para Baixo/efeitos dos fármacos , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Genes bcl-2 , Células HeLa/efeitos dos fármacos , Células HeLa/metabolismo , Células HeLa/patologia , Humanos , Mifepristona/farmacologia , Proteínas de Neoplasias/genética , Proteínas de Neoplasias/metabolismo , Poli(ADP-Ribose) Polimerases/metabolismo , Receptores de Glucocorticoides/efeitos dos fármacos , Fator de Transcrição AP-1/metabolismo , Transcrição Gênica/efeitos dos fármacos , Triancinolona/farmacologia , Ácido UrsólicoRESUMO
Bipolar disorder (BD) is characterized by hypothalamic pituitary adrenal (HPA) axis hyperactivity, glucocorticoid insensitivity and alterations in serotonin and inflammatory mediators. The glucocorticoid receptor (GR), activator protein-1 (AP-1), nuclear factor-kappa B (NF-kappaB) and c-jun N-terminal kinase (JNK) regulate the above mentioned processes; we therefore assessed their role in BD. Fifteen bipolar depressed patients under multiple anti-depressant therapy, 15 bipolar euthymics under lithium monotherapy and 25 matched controls were studied. Whole cell and nuclear extracts from lymphocytes were immunoblotted for GR, c-fos, JNK and NF-kappaB and nuclear aliquots were submitted to electrophoretic mobility shift assay for GR, AP-1 and NF-kappaB. Associations with the anti-depressant therapy and the state of the disease were also sought. Results, expressed as percentage of pooled protein standard sample intergraded optical density (IOD) (mean +/- SD), revealed: (a) depressed patients had significantly higher GR levels than controls in whole cell (82.63 +/- 6.18 versus 76.27 +/- 4.21%, P < 0.01) and nuclear extracts (86.66 +/- 3.81 versus 81.72 +/- 2.71%, P < 0.001) but lower GR-DNA binding (68.75 +/- 7.91 versus 81.84 +/- 4.25%, P < 0.05). Euthymics had normalized whole cell GR content (73.64 +/- 5.95%) and GR-DNA binding activity (76.82 +/- 7.29%) but higher nuclear GR content (86.89+/-3.96%, P<0.01) than controls; (b) nuclear c-fos content and AP-1-DNA-binding were significantly lower in depressed patients than controls (80.49 +/- 2.03 versus 84.82 +/- 3.48%, P < 0.05 and 78.46 +/- 4.17 versus 84.80 +/- 5.79%, P < 0.05, respectively). Euthymics however, showed similar nuclear c-fos and AP-1-DNA-binding to controls (85.48 +/- 2.71 and 87.78 +/- 3.54%, respectively) but lower whole cell c-fos than in controls (81.18 +/- 3.87 versus 87.01 +/- 4.22%, P < 0.001); (c) depressed patients had significantly lower whole cell and nuclear JNK than controls (67.01 +/- 4.29 versus 72.00 +/- 3.68%, P < 0.05 and 80.10 +/- 2.53 versus 86.96 +/- 2.49%, P < 0.001) whereas euthymics showed lower nuclear JNK (83.27 +/- 1.93%, P < 0.01); (d) whole cell NF-kB was higher in the depressed patients than in controls (67.30 +/- 5.00 versus 63.63 +/- 3.3%, P < 0.05). Concluding, intracellular signaling of GR, AP-1 and JNK are altered in BD and may underly disease aetiopathogenesis and/or reflect the effect of the anti-depressants.