RESUMEN
BACKGROUND: Epithelial-to-Mesenchymal Transition (EMT) is necessary for metastasis. Zinc- finger domain-containing transcription factors, especially Snail1, bind to E-box motifs and play a crucial role in the induction and regulation of EMT. OBJECTIVE: We hypothesized if C-terminal region of Snail1 (CSnail1) may competitively bind to E-box and block cancer metastasis. METHODS: The CSnail1 gene coding sequence was inserted into the pIRES2-EGFP vector. Following transfection of A549 cells with the designed construct, EMT was induced with TGF-ß1 and the expression of essential EMT markers was evaluated by real-time PCR and immunoblotting. We also monitored cell migration. RESULTS: CSnail1 inhibited TGF-ß1-induced N-cadherin and vimentin mRNA expression and increased ß-catenin expression in transfected TGF-ß1-treated A549 cells. A similar finding was obtained in western blotting. CSnail1 also blocked the migration of transfected cells in the scratch test. CONCLUSION: Transfection of A549 cells with CSnail1 alters the expression of essential EMT markers and consequently suppresses tumor cell migration. These findings confirm the capability of CSnail1 in EMT blocking and in parallel to current patents could be applied as a novel strategy in the prevention of metastasis.
Asunto(s)
Adenocarcinoma Bronquioloalveolar/genética , Biomarcadores de Tumor/genética , Movimiento Celular/genética , Transición Epitelial-Mesenquimal/genética , Neoplasias Pulmonares/genética , Factores de Transcripción de la Familia Snail/fisiología , Células A549 , Adenocarcinoma Bronquioloalveolar/patología , Movimiento Celular/efectos de los fármacos , Codón sin Sentido , Transición Epitelial-Mesenquimal/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Regulación Neoplásica de la Expresión Génica/genética , Humanos , Neoplasias Pulmonares/patología , Dominios Proteicos/genética , Dominios Proteicos/fisiología , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/farmacología , Factores de Transcripción de la Familia Snail/química , Factores de Transcripción de la Familia Snail/genética , Factores de Transcripción de la Familia Snail/farmacología , Factor de Crecimiento Transformador beta1/farmacologíaRESUMEN
Sleep is considered as a physiological regulator in the body. Gamma-aminobutyric acid (GABA) is a neurotransmitter that modulates sleep and affects cardiac functions. We evaluated effects of acute sleep deprivation (SD) on cardiac hemodynamic parameters, expression of pro-inflammatory cytokines, and Heat shock protein (Hsp70), serum level of lactate dehydrogenase (LDH) and prooxidant/antioxidant balance (PAB). Male Wistar rats were bilaterally cannulated in the central nucleus of amygdala (CeA) and saline or bicuculline was injected 24 hours prior to induction of 30 minute ischemia following 120 minute reperfusion. Forty-eight animals were randomly divided into four groups: Control (CONT), bicuculline (BIC), acute SD and bicuculline + acute sleep deprivation (BIC+SD). Animals in SD and BIC+SD groups were put in an aquarium for inducing sleep deprivation. SD attenuated LDH, pro-inflammatory cytokines and PAB; improved cardiac hemodynamic parameters and increased Hsp70 in non-infarcted area as compared to CONT. Administration of bicuculline increased LDH, pro-inflammatory cytokines and PAB, reduced cardiac hemodynamic parameters and Hsp70 as compared to CONT. Furthermore, bicuculline administration prior to acute sleep induction decreased SD effects on LDH, PAB, Hsp70, cardiac hemodynamic parameters and pro-inflammatory cytokines. Induction of SD prior to ischemia/reperfusion induces cardioprotection through suppressing inflammatory responses.
Asunto(s)
Corazón/fisiopatología , Miocardio/metabolismo , Receptores de GABA-A/metabolismo , Daño por Reperfusión/prevención & control , Privación de Sueño/metabolismo , Animales , Antioxidantes/metabolismo , Regulación de la Expresión Génica , Proteínas HSP70 de Choque Térmico/genética , Hemodinámica , Inflamación/metabolismo , Interleucina-6/genética , L-Lactato Deshidrogenasa/metabolismo , Masculino , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/metabolismo , Daño por Reperfusión/metabolismo , Daño por Reperfusión/fisiopatología , Privación de Sueño/fisiopatología , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
Common variable immunodeficiency (CVID) is a heterogeneous group of disorders, characterized by hypogammaglobulinemia and normal or low numbers of B cells, which predispose patients to recurrent infections. Peripheral blood mononuclear cells from 19 patients with CVID, and age- and sex-matched controls, were subjected to an in vitro assay of B-cell-T-cell activation and interaction, using anti-immunoglobulin (Ig)-D conjugated to dextran (alpha-delta-dex), as a polyclonal T independent type 2 antigen mimic, with and without anti-CD3/anti-CD28, as polyclonal T-cell stimuli. Stimulation of lymphocytes with either anti-CD3 or anti-CD3 plus anti-CD28 induced T-cell activation and proliferation in CVID patients who were similar to age- and sex-matched controls, but B cells of patients were significantly less activated when peripheral blood mononuclear cells were stimulated with polyclonal T-cell agonists alone. Comparison of CD86 expression in the patients with matched controls revealed that patients had low B-cell activation in response to T-cell stimuli (bystander T-cell help). In conclusion, this sample of CVID patients exhibits a defect of T-cell "help" to B cells, and/or B-cell response to T-cell help.