RESUMEN
Background/aim: In the present study we aimed to figure out the effect of metformin on the expression of AMPK-alpha, cyclin D1, and Tp53, and apoptosis in primary breast cancer cells (PBCCs). Materials and methods: PBCCs were treated with two doses of metformin (0 mM, 25 mM). Proliferation was determined by BrdU as- say. Real-time PCR was used to assess AMPK-alpha, cyclin D1, and Tp53 gene expressions; apoptotic indexes of PBCCs were analyzed using flow-cytometry. Results: Twenty-fourhour incubation with 25 mM metformin reduced the proliferation of PBCCs. AMPK-alpha gene expression in PBCCs was not affected by 25 mM metformin treatment compared with the control group. PBCCs treated with 25 mM metformin had lower cyclin D1 expression compared with nontreated cells; however, the difference was not statistically significant. Twenty-five mil- limolar dose of metformin increased p53 expression significantly compared with the nontreated group. The high concentration of met- formin elevated the number of annexin V-positive apoptotic cells, and the increase in the apoptotic index was statistically significant. Conclusion: Metformin can modulate cyclin D1 and p53 expression through AMPK-alpha-independent mechanism in breast cancer cells, leading to cell proliferation inhibition and apoptosis induction.
Asunto(s)
Proteínas Quinasas Activadas por AMP/metabolismo , Apoptosis/efectos de los fármacos , Neoplasias de la Mama/tratamiento farmacológico , Proliferación Celular/efectos de los fármacos , Ciclina D1/metabolismo , Hipoglucemiantes/farmacología , Metformina/farmacología , Proteína p53 Supresora de Tumor/metabolismo , Proteínas Quinasas Activadas por AMP/genética , Apoptosis/fisiología , Neoplasias de la Mama/genética , Línea Celular Tumoral , Ciclina D1/genética , Regulación hacia Abajo , Femenino , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Humanos , Persona de Mediana Edad , Proteína p53 Supresora de Tumor/genética , Regulación hacia ArribaRESUMEN
Alcohol abuse is a well-known cause of imbalance in trace element levels and oxidant/antioxidant status of individuals with long time consumption. However, the levels of these parameters in the patients on the early stages of alcohol dependence without liver damage differ on various studies. The aim of our study was to measure the levels of trace elements in the serum and oxidative/antioxidative system members in the red blood cells (RBC) of early-stage alcoholic individuals and compare with control subjects. Our study included 21 male patients recently hospitalized for alcohol abuse and 25 healthy non-abusing male controls. Levels of Fe, Zn, and Cu in the serum and MDA, SOD, CAT, and GSH in the red blood cells (RBC) of the subjects were measured. Fe, Zn, and Cu levels were lower in the study group when compared to the controls. Levels of lipid peroxidation marker MDA was high, whereas the activities of antioxidant enzymes SOD and CAT were decreased in our study group. However, levels of GSH, an antioxidant compound were higher in the alcohol abuse group. RBC SOD levels were positively correlated with Fe, Cu, Zn, and CAT. There was a positive correlation between Fe-Cu, Zn-Fe, Zn-Cu, CAT-Zn, and CAT-SOD. MDA was negatively correlated with Fe, Zn, SOD, and CAT. The results obtained from present study indicate that high levels of alcohol intake are related with increased oxidative damage and decreased levels of antioxidant enzymes and trace elements. Additionally, antioxidant compensation mechanisms are still on process in the early stages of chronic alcohol exposure.