Analysis of stemness gene expression and CD133 abnormal methylation in neuroblastoma cell lines

  • Authors:
    • Paula Schiapparelli
    • Mónica Enguita-Germán
    • Jana Balbuena
    • Juan A. Rey
    • Paula Lázcoz
    • Javier S. Castresana
  • View Affiliations

  • Published online on: November 1, 2010     https://doi.org/10.3892/or_00000993
  • Pages: 1355-1362
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Neuroblastoma is the most common extracranial solid tumor in children, accounting for up to 10% of all childhood malignancies. Cellular heterogeneity is a hallmark of this embryonal cancer, as distinct neural crest lineages can be found within the same tumor sample. The aim of our study was to investigate the presence of a subpopulation of immature cells with features of cancer-like stem cells in 10 neuroblastoma cell lines. RT-PCR and flow cytometry were performed in order to analyze different kinds of ‘stemness genes’ such as: NESTIN (NES), CD133, SOX-2, BMI1, c-KIT, MELK1, MUSASHI-1 (MSI1), FAS, CD44 and VIMENTIN (VIM). In addition, glial and neuronal markers such as NCAM1, GFAP and B-TUBULIN III (TUBB3) were analyzed. Epigenetic changes within the CD133 (Prominin-1) gene promoter were also analyzed. Neuroblastoma cell lines showed a particular pattern of expression, suggesting the presence of an immature cancer stem cell-like subpopulation. The CD133 protein, commonly used to enrich putative cancer propagating stem cell-like populations in different kinds of solid tumors, presented a half-methylated DNA state in 7 of the 12 neuroblastoma cell lines analyzed. An increase in RNA and protein levels of CD133 was achieved following demethylation by assays using 5-aza-2'-deoxycytidine (5-Aza-dC). Since cancer stem cells are believed to be responsible for tumor metastasis, escape from anticancer therapies and disease relapse, their therapeutic targeting and analysis is crucial in neuroblastoma. Moreover, the regulation of CD133 by epigenetic changes may provide an innovative mechanism of CD133 expression as its regulation still remains unclear.

Related Articles

Journal Cover

November 2010
Volume 24 Issue 5

Print ISSN: 1021-335X
Online ISSN:1791-2431

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Schiapparelli P, Enguita-Germán M, Balbuena J, Rey JA, Lázcoz P and Castresana JS: Analysis of stemness gene expression and CD133 abnormal methylation in neuroblastoma cell lines . Oncol Rep 24: 1355-1362, 2010
APA
Schiapparelli, P., Enguita-Germán, M., Balbuena, J., Rey, J.A., Lázcoz, P., & Castresana, J.S. (2010). Analysis of stemness gene expression and CD133 abnormal methylation in neuroblastoma cell lines . Oncology Reports, 24, 1355-1362. https://doi.org/10.3892/or_00000993
MLA
Schiapparelli, P., Enguita-Germán, M., Balbuena, J., Rey, J. A., Lázcoz, P., Castresana, J. S."Analysis of stemness gene expression and CD133 abnormal methylation in neuroblastoma cell lines ". Oncology Reports 24.5 (2010): 1355-1362.
Chicago
Schiapparelli, P., Enguita-Germán, M., Balbuena, J., Rey, J. A., Lázcoz, P., Castresana, J. S."Analysis of stemness gene expression and CD133 abnormal methylation in neuroblastoma cell lines ". Oncology Reports 24, no. 5 (2010): 1355-1362. https://doi.org/10.3892/or_00000993