Background: Spontaneous regression is usually found in stage 4s neuroblastoma, whereas the elucidation of the underlying molecular mechanism(s) is still limited. Purpose: Our study aims to investigate the pathogenesis of spontaneous regression at the protein level. Methods and materials: Differential expression of proteins in stage 4s neuroblastoma tissue, in stage 4 neuroblastoma tissue, and in normal adrenal tissue was investigated by use of 2-dimensional difference gel electrophoresis (2D-DIGE). Results: Twenty-four protein spots were found to have significant changes among the different tissues, in which 16 proteins were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Among these proteins, 7 proteins (RhoGDP-dissociation inhibitor 1, phosphatidylethanolamine-binding protein, prohibitin, etc) were up-regulated and 2 proteins (F-actin capping protein 1 subunit and aldose reductase) were down-regulated in stage 4s neuroblastoma compared with stage 4 neuroblastoma. The differential expression of selected candidate protein (RhoGDP-dissociation inhibitor 1 and CAPZA1) was further validated by western blotting. Conclusion: Some proteins are differentially expressed between stage 4s and stage 4 neuroblastoma tissue, including those associated with differentiation and proliferation as well as apoptosis. RhoGDP-dissociation inhibitor 1 is highly expressed in stage 4s neuroblastoma tissue, whereas CAPZA1 is down-regulated. (C) 2011 Elsevier Inc. All rights reserved.