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1.
Braz. j. med. biol. res ; 50(1): e5794, 2017. graf
Artículo en Inglés | LILACS | ID: biblio-839241

RESUMEN

Propofol is a frequently used intravenous anesthetic agent. Recent studies show that propofol exerts a number of non-anesthetic effects. The present study aimed to investigate the effects of propofol on lung cancer cell lines H1299 and H1792 and functional role of microRNA (miR)-486 in these effects. H1299 and/or H1792 cells were treated with or without propofol and transfected or not with miR-486 inhibitor, and then cell viability and apoptosis were analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and flow cytometry. The expression of miR-486 was determined by quantitative real-time polymerase chain reaction (qRT-PCR) with or without propofol treatment. Western blot was performed to analyze the protein expression of Forkhead box, class O (FOXO) 1 and 3, Bcl-2 interacting mediator of cell death (Bim), and pro- and activated caspases-3. Results showed that propofol significantly increased the miR-486 levels in both H1299 and H1792 cells compared to untreated cells in a dose-dependent manner (P<0.05 or P<0.01). Propofol statistically decreased cell viability but increased the percentages of apoptotic cells and protein expressions of FOXO1, FOXO3, Bim, and pro- and activated caspases-3; however, miR-486 inhibitor reversed the effects of propofol on cell viability, apoptosis, and protein expression (P<0.05 or P<0.01). In conclusion, propofol might be an ideal anesthetic for lung cancer surgery by effectively inhibiting lung cancer cell viability and inducing cell apoptosis. Modulation of miR-486 might contribute to the anti-tumor activity of propofol.


Asunto(s)
Humanos , Neoplasias Pulmonares/metabolismo , MicroARNs/metabolismo , Propofol/farmacología , Apoptosis/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Reacción en Cadena en Tiempo Real de la Polimerasa
2.
Braz. j. med. biol. res ; 48(4): 286-291, 4/2015. tab, graf
Artículo en Inglés | LILACS | ID: lil-744366

RESUMEN

This study aimed to determine the effects of different concentrations of propofol (2,6-diisopropylphenol) on lipopolysaccharide (LPS)-induced expression and release of high-mobility group box 1 protein (HMGB1) in mouse macrophages. Mouse macrophage cell line RAW264.7 cells were randomly divided into 5 treatment groups. Expression levels of HMGB1 mRNA were detected using RT-PCR, and cell culture supernatant HMGB1 protein levels were detected using enzyme-linked immunosorbent assay (ELISA). Translocation of HMGB1 from the nucleus to the cytoplasm in macrophages was observed by Western blotting and activity of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) in the nucleus was detected using ELISA. HMGB1 mRNA expression levels increased significantly in the cell culture supernatant and in cells after 24 h of stimulating RAW264.7 cells with LPS (500 ng/mL). However, HMGB1 mRNA expression levels in the P2 and P3 groups, which received 500 ng/mL LPS with 25 or 50 μmol/mL propofol, respectively, were significantly lower than those in the group receiving LPS stimulation (P<0.05). After stimulation by LPS, HMGB1 protein levels were reduced significantly in the nucleus but were increased in the cytoplasm (P<0.05). Simultaneously, the activity of NF-κB was enhanced significantly (P<0.05). After propofol intervention, HMGB1 translocation from the nucleus to the cytoplasm and NF-κB activity were inhibited significantly (each P<0.05). Thus, propofol can inhibit the LPS-induced expression and release of HMGB1 by inhibiting HMGB1 translocation and NF-κB activity in RAW264.7 cells, suggesting propofol may be protective in patients with sepsis.


Asunto(s)
Animales , Ratones , Anestésicos Intravenosos/farmacología , Núcleo Celular/efectos de los fármacos , Proteína HMGB1/efectos de los fármacos , Macrófagos/efectos de los fármacos , Propofol/farmacología , ARN Mensajero/efectos de los fármacos , Transporte Activo de Núcleo Celular , Anestésicos Intravenosos/administración & dosificación , Western Blotting , Línea Celular , Núcleo Celular/metabolismo , Ensayo de Inmunoadsorción Enzimática , Expresión Génica/efectos de los fármacos , Proteína HMGB1/genética , Proteína HMGB1/metabolismo , Lipopolisacáridos , Macrófagos/metabolismo , FN-kappa B/efectos de los fármacos , FN-kappa B/metabolismo , Propofol/administración & dosificación , Distribución Aleatoria , Reacción en Cadena en Tiempo Real de la Polimerasa , ARN Mensajero/metabolismo
3.
Braz. j. med. biol. res ; 47(3): 252-258, 03/2014. tab, graf
Artículo en Inglés | LILACS | ID: lil-704618

RESUMEN

Beclin 1 plays a critical role in autophagy and functions as a haploinsufficient tumor suppressor. The expression and prognostic significance of beclin 1 in head and neck adenoid cystic carcinoma (ACC) are largely unexplored. Therefore, we investigated the expression of beclin 1, Bcl-2, and p53 in head and neck ACC tissue. Tissue samples from 35 cases (15 females, 20 males) of head and neck ACC were utilized for immunohistochemistry. Beclin 1 expression was observed in 32 cases (91.4%) and considered to be high in 15 cases (42.9%) and low in 20 cases (57.1%). Beclin 1 expression was significantly correlated with a histological growth pattern (P=0.046) and histological grade (P=0.037). Beclin 1 expression was inversely correlated with Bcl-2 expression (P=0.013) and significantly associated with overall survival (P=0.006). Bcl-2 and p53 expression were observed in 21 cases (60.0%) and 16 cases (45.7%). Bcl-2 expression was significantly correlated with perineural invasion (P=0.041) and not associated with overall survival (P=0.053). p53 expression was directly correlated with beclin 1 expression (P=0.044). Our results indicated that beclin 1 may be a novel, promising prognostic factor for clinical outcome in head and neck ACC patients and may play a part in the development of head and neck ACC by interacting with Bcl-2 and p53.


Asunto(s)
Adolescente , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Proteínas Reguladoras de la Apoptosis/metabolismo , Carcinoma Adenoide Quístico/metabolismo , Proteínas de la Membrana/metabolismo , /metabolismo , Neoplasias de las Glándulas Salivales/metabolismo , /análisis , Autofagia/fisiología , Neoplasias de Cabeza y Cuello/metabolismo , Inmunohistoquímica , Estimación de Kaplan-Meier , Pronóstico
4.
Braz. j. med. biol. res ; 43(1): 43-51, Jan. 2010. ilus, tab
Artículo en Inglés | LILACS | ID: lil-535635

RESUMEN

Myocardial ischemic preconditioning up-regulated protein 1 (Mipu1), a novel zinc finger protein, was originally cloned using bioinformatic analysis and 5' RACE technology of rat heart after a transient myocardial ischemia/reperfusion procedure in our laboratory. In order to investigate the functions of Mipu1, the recombinant prokaryotic expression vector pQE31-Mipu1 was constructed and transformed into Escherichia coli M15(pREP4), and Mipu1-6His fusion protein was expressed and purified. The identity of the purified protein was confirmed by mass spectrometry. The molecular mass of the Mipu1 protein was 70.03779 kDa. The fusion protein was intracutaneously injected to immunize New Zealand rabbits to produce a polyclonal antibody. The antibody titer was approximately 1:16,000. The antibody was tested by Western blotting for specificity and sensitivity. Using the antibody, it was found that Mipu1 was highly expressed in the heart and brain of rats and was localized in the nucleus of H9c2 myogenic cells. The present study lays the foundation for further study of the biological functions of Mipu1.


Asunto(s)
Animales , Conejos , Ratas , Anticuerpos Monoclonales/biosíntesis , Química Encefálica , Isquemia Miocárdica/genética , Miocardio/química , Proteínas Nucleares/metabolismo , Proteínas Represoras/metabolismo , Clonación Molecular , Escherichia coli/genética , Regulación de la Expresión Génica , Vectores Genéticos/genética , Vectores Genéticos/metabolismo , Espectrometría de Masas , Reperfusión Miocárdica , Proteínas Nucleares/genética , Proteínas Represoras/genética , Sensibilidad y Especificidad , Transfección
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