Contributed equally to this work.
Identification of novel genes associated with astrocytoma progression using suppression subtractive hybridization and real-time reverse transcription-polymerase chain reaction
Version of Record online: 24 JUL 2006
Copyright © 2006 Wiley-Liss, Inc.
International Journal of Cancer
Volume 119, Issue 10, pages 2330–2338, 15 November 2006
How to Cite
van den Boom, J., Wolter, M., Blaschke, B., Knobbe, C. B. and Reifenberger, G. (2006), Identification of novel genes associated with astrocytoma progression using suppression subtractive hybridization and real-time reverse transcription-polymerase chain reaction. Int. J. Cancer, 119: 2330–2338. doi: 10.1002/ijc.22108
- Issue online: 26 SEP 2006
- Version of Record online: 24 JUL 2006
- Manuscript Accepted: 3 MAY 2006
- Manuscript Received: 8 FEB 2006
- Deutsche Forschungsgemeinschaft. Grant Number: SFB503-B7
- Deutsche Krebshilfe. Grant Numbers: 70-3088-Sa1, 70-3163-Wi3
- German Bundesministerium für Bildung und Forschung (BMBF) within the National Genome Research Network 2 (NGFN-2)
- Neuroscience Research Training Group at Heinrich-Heine-University
- adhesion molecule on glia;
- suppression subtractive hybridization;
- tumor progression
To identify novel genes involved in glioma progression we performed suppression subtractive hybridization combined with cDNA array analysis on 4 patients with primary low-grade gliomas of World Health Organization (WHO) grade II that recurred as secondary glioblastomas (WHO grade IV). Eight genes showing differential expression between primary and recurrent tumors in 3 of the 4 patients were selected for further analysis using real-time reverse transcription-PCR on a series of 10 pairs of primary low-grade and recurrent high-grade gliomas as well as 42 astrocytic gliomas of different WHO grades. These analyses revealed that 5 genes, i.e., AMOG (ATP1B2, 17p13.1), APOD (3q26.2-qter), DMXL1 (5q23.1) DRR1 (TU3A, 3p14.2) and PSD3 (KIAA09428/HCA67/EFA6R, 8p22), were expressed at significantly lower levels in secondary glioblastomas as compared to diffuse astrocytomas of WHO grade II. In addition, AMOG, DRR1 and PSD3 transcript levels were significantly lower in primary glioblastomas than in diffuse astrocytomas. Treatment of glioma cell lines with 5-aza-2′-deoxycytidine and trichostatin A resulted in increased expression of AMOG and APOD transcripts. Sequencing of sodium bisulfite-modified DNA demonstrated AMOG promoter hypermethylation in the glioma cell lines and 1 primary anaplastic astrocytoma with low AMOG expression. Taken together, we identified interesting novel candidate genes that likely contribute to glioma progression and provide first evidence for a role of epigenetic silencing of AMOG in malignant glioma cells. © 2006 Wiley-Liss, Inc.