The role of nerve growth factor in caspase-dependent apoptosis in human BE(2)C neuroblastoma

Purpose: The purpose of the study was to determine if nerve growth factor (NGF) stimulation induces apoptosis in the BE(2) C neuroblastoma cell line in vitro. Methods: The LPCX retroviral vector was used to achieve stable transduction of NGF complementary DNA into BE(2) C neuroblastoma cells. Wild-type and NGF-transduced cells were then incubated with varying concentrations of NGF for varying periods. A laddering assay was performed to determine the presence of DNA fragments characteristic of apoptosis. The expression of various cleaved and total caspases was determined by Western immunoblotting. Results: p75 receptor expression in the NGF-transduced cell line was equivalent to that in the wild-type cell line, but Trk A receptor expression was significantly decreased in BE(2)C-NGF cells. DNA laddering assay demonstrated that only BE(2) C-NGF cells underwent apoptosis after stimulation with exogenous NGF. BE(2) C-NGF cells have increased expression of cleaved caspase-3 when compared with wild-type cells. Cleaved caspase-3 expression is further increased with exogenous NGF stimulation in the transduced cells. Conclusion: This study confirms that NGF stimulation of BE(2) C neuroblastoma cells can induce apoptosis through activation of the caspase cascade in vitro. The differential expression of the receptors Trk A and p75 between the wild-type and NGF-transduced cell lines may explain the differing effects observed. (C) 2011 Elsevier Inc. All rights reserved.