Original Article
Proteomics-based identification of spontaneous regression-associated proteins in neuroblastoma

https://doi.org/10.1016/j.jpedsurg.2011.06.024Get rights and content

Abstract

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.

Section snippets

Patients and clinical samples

Tumor tissue samples from 8 patients who were all younger than 1 year and had been diagnosed with neuroblastoma were obtained from the Beijing Children's Hospital and China PLA General Hospital. Among these cases, 4 were selected as stage 4 neuroblastoma and the other 4 as stage 4s neuroblastoma. The distinction of tumor staging was based on the International Neuroblastoma Staging System. Tumor resections were performed after valid informed consent had been taken. The samples were collected at

Separation of total proteins by 2D-DIGE

Although the internal standard was incorporated into the 2D-DIGE experiment to eliminate the gel-to-gel variation, good reproducibility among different gels is the basis for effective differential protein expression analysis. The Cy2-labeled gel images of the 4 gels on which the same internal standard was loaded in equal amounts displayed a good reproducibility among different gels and was significantly elevated overall in our experiments.

The 3 cyanine dye images for each gel were overlaid.

Discussion

The mechanism for spontaneous regression of neuroblastomas is still poorly understood. Some researchers believed that neuroblastoma tissue was derived from a malignant transformation of degenerated embryonic tissue; therefore, its spontaneous regression might be, in fact, regression of the embryo. Recently, some groups found morphologic features of apoptosis in stage 4s neuroblastomas, which led to the assumption that apoptosis, which is a representative form of programmed cell death (PCD),

Acknowledgments

This research work was fully supported by a grant from the Chinese Natural Science Foundation, Permit No. 30772275, and a grant from China Project 863, Permit No. 2007AA021004.

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