Epidermal growth factor receptor is required for estradiol-stimulated bovine satellite cell proliferation

The objective of this study was to assess the role of the epidermal growth factor receptor (EGFR) in estradiol-17 beta (E-2)-stimulated proliferation of cultured bovine satellite cells (BSCs). Treatment of BSC cultures with AG1478 (a specific inhibitor of EGFR tyrosine kinase activity) suppresses E-2-stimulated BSC proliferation (P < 0.05). In addition, E-2-stimulated proliferation is completely suppressed (P < 0.05) in BSCs in which EGFR expression is silenced by treatment with EGFR small interfering RNA (siRNA). These results indicate that EGFR is required for E2 to stimulate proliferation in BSC cultures. Both AG1478 treatment and EGFR silencing also suppress proliferation stimulated by LR3-IGF-1 (an IGF1 analogue that binds normally to the insulin-like growth factor receptor (IGFR)-1 but has little or no affinity for IGF binding proteins) in cultured BSCs (P < 0.05). Even though EGFR siRNA treatment has no effect on IGFR-1 beta mRNA expression in cultured BSCs, IGFR-1 beta protein level is substantially reduced in BSCs treated with EGFR siRNA. These data suggest that EGFR silencing results in post-transcriptional modifications that result in decreased IGFR-1 beta protein levels. Although it is clear that functional EGFR is necessary for E2-stimulated proliferation of BSCs, the role of EGFR is not clear. Transactivation of EGFR may directly stimulate proliferation, or EGFR may function to maintain the level of IGFR-1 beta which is necessary for E2-stimulated proliferation. It also is possible that the role of EGFR in E-2-stimulated BSC proliferation may involve both of these mechanisms. (C) 2014 Elsevier Inc. All rights reserved.