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Ann. hepatol ; 16(2): 297-303, Mar.-Apr. 2017. tab, graf
Artículo en Inglés | LILACS | ID: biblio-887236

RESUMEN

ABSTRACT Introduction and aim. The inability to distinguish cancer (CSCs) from normal stem cells (NSCs) has hindered attempts to identify safer, more effective therapies for hepatocellular carcinoma (HCC). The aim of this study was to document and compare cell membrane potential differences (PDs) of CSCs and NSCs derived from human HCC and healthy livers respectively and determine whether altered GABAergic innervation could explain the differences. Material and methods. Epithelial cell adhesion molecule (EpCAM) positive stem cells were isolated from human liver tissues by magnetic bead separations. Cellular PDs were recorded by microelectrode impalement of freshly isolated cells. GABAA receptor subunit expression was documented by reverse transcriptase polymerase chain reaction (RT-PCR) and immunofluorescence. Results. CSCs were significantly depolarized (-7.0 ± 1.3 mV) relative to NSCs (-23.0 ± 1.4 mV, p < 0.01). The depolarized state was associated with different GABAA receptor subunit expression profiles wherein phasic transmission, represented by GAGAA α3 subunit expression, was prevalent in CSCs while tonic transmission, represented by GABAA α6 subunit expression, prevailed in NSCs. In addition, GABAA subunits α3, β3, γ3 and δ were strongly expressed in CSCs while GABAA π expression was dominant in NSCs. CSCs and NSCs responded similarly to GABAA receptor agonists (ΔPD: 12.5 ± 1.2 mV and 11.0 ± 3.5 mV respectively). Conclusion. The results of this study indicate that CSCs are significantly depolarized relative to NSCs and these differences are associated with differences in GABAA receptor subunit expression. Together they provide new insights into the pathogenesis and possible treatment of human HCC.


Asunto(s)
Humanos , Células Madre Neoplásicas/metabolismo , Receptores de GABA-A/metabolismo , Carcinoma Hepatocelular/genética , Carcinoma Hepatocelular/metabolismo , Agonistas de Receptores de GABA-A/farmacología , Molécula de Adhesión Celular Epitelial/metabolismo , Hígado/citología , Neoplasias Hepáticas/metabolismo , Fenotipo , Células Madre/efectos de los fármacos , Células Madre Neoplásicas/efectos de los fármacos , ARN Mensajero/genética , ARN Mensajero/metabolismo , Biomarcadores/metabolismo , Técnica del Anticuerpo Fluorescente , Separación Inmunomagnética , Receptores de GABA-A/efectos de los fármacos , Receptores de GABA-A/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Subunidades de Proteína , Neoplasias Hepáticas/genética , Potenciales de la Membrana/efectos de los fármacos
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