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2014, Número 4

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Biotecnol Apl 2014; 31 (4)


Altas concentraciones de glucosa inhiben la cascada de señalización EGFR/PI3K/AKT1/mTOR en fibroblastos cutáneos

Mendoza-Marí Y, Garcia-Ojalvo A, Fernández-Mayola M, Herrera-Martínez L, Berlanga-Acosta J

Idioma: Ingles.
Referencias bibliográficas: 56
Paginas: 285-290
Archivo PDF: 704.94 Kb.


RESUMEN

La fisiología de los fibroblastos, esenciales para la cicatrización, se perturba ante altas concentraciones de glucosa. Se estudió el efecto de altas cargas de glucosa sobre la proliferación de fibroblastos cutáneos de donantes sanos a través de la cascada de señalización del factor de crecimiento epidérmico (EGFR): la autofosforilación del EGFR y la activación de intermediarios de la transducción hasta la ciclina D1. Los fibroblastos se cultivaron durante seis días en condiciones estándar con SFB al 15 %, y glucosa a 5 mM (glucosa normal) o 35 mM (glucosa alta). Se incluyó un control osmótico concurrente. Tras seis días se estimó el tiempo de duplicación. Las suspensiones celulares se sembraron, fijaron e inmunomarcaron con anticuerpos específicos contra las formas fosforiladas de EGFR (Y1197), AKT1 (S473) y mTOR (S2448), y las formas nativas de la subunidad p85 alfa de la PI3K y la ciclina D1. Se calculó la proporción de células positivas a la reacción de diaminobenzidina/peroxidasa, y su intensidad según procedimientos estándar. Las altas concentraciones de glucosa multiplicaron en 5 veces el tiempo de duplicación comparado con el de células cultivadas en condiciones fisiológicas. También redujo la autofosforilación constitutiva del EGFR, atenuó significativamente la subsiguiente fosforilación de PI3K y muy significativamente la de sus intermediarios de transducción AKT1 y mTOR. La expresión de la ciclina D1 se abolió ante las altas cargas de glucosa. Los resultados sugieren que la exposición a altas concentraciones de glucosa afecta la proliferación de los fibroblastos al perturbar el eje de transducción EGFR/PI3K/AKT1/mTOR/Ciclina D1.


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