Diadenosine polyphosphates, Ap₄A and Ap₃A, increase epithelial cell proliferation during corneal wound healing

• Autores: Aránzazu Mediero Muñoz, Almudena Crooke, Jesús Pintor
• Localización: Journal of Emmetropia: Journal of Cataract, Refractive and Corneal Surgery, ISSN-e 2171-4703, Vol. 1, Nº. 2, 2010, págs. 81-87
• Idioma: inglés
• Texto completo no disponible (Saber más ...)
• Resumen

  • PURPOSE: To study the effect of Ap₄A and Ap₃A on corneal epithelial cell proliferation during the wound healing process.

    METHODS: In vitro proliferation assays were performed in SIRC cells studying proliferation at 24 and 36 hours by means of CyQUANT® NF Cell Proliferation Assay when cells were challenged with diadenosine polyphosphates. In vivo, proliferation assay was performed by measuring the relative quantification of the cell cyclins CCNE1 and CDK2 by means of qRT-PCR at 24 and 36 hours after the wounds were performed.

    RESULTS: In vitro, both Ap₄A and Ap₃A produced an increase in cell proliferation 24 and 36 hours after the first incubation, this being reduced in the presence of all tested inhibitors. The pre-treatment with one dinucleotide in the presence of the other produced a high increase in the proliferation rate. qRT-PCR assay for the activation of the cell cyclins CCNE1/CDK2 system, revealed that Ap₄A increased mRNA expression for CCNE1 and decresed CDK2 levels. In the presence of Ap₃A, CCNE1 and CDK2 mRNA expression after 24 hours of the wound was increased in a significant manner related to control expression.

    Pre-treatment with P2Y₂ siRNA produced a decrease in the cyclins levels.

    CONCLUSIONS: Ap₄A produced its effect on proliferation by activation of a P2Y₂ receptor coupled to canonical PLC/PKC/RhoA/ROCK-I pathways plus the transactivation of a tyrosin kinase receptor. On the other hand, ApA produced an increase in proliferation by activation of Erk1/2 and p38 pathways by a means of a P2Y₆ receptor