| Autor: |
Barbara J. Lamb, Rork Kuick, Peter F. Ambros, Markus Eckart, Andreas Weinhäusl, Xiao-Xiang Zhu, Katharina Wimmer, Jean Marie Rouillard, Christa Fonatsch, Sam M. Hanash |
| Rok vydání: |
2002 |
| Předmět: |
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| Zdroj: |
Genes, Chromosomes and Cancer. 33:285-294 |
| ISSN: |
1045-2257 |
| DOI: |
10.1002/gcc.10030 |
| Popis: |
Restriction landmark genome scanning (RLGS) allows comparative analysis of several thousand DNA fragments in the genome and provides a means to identify CpG islands that are altered in tumor cells as a result of amplification, deletion, or methylation changes. We have developed a novel informatics tool, designated virtual genome scan (VGS), that makes it possible to predict automatically the sequence of fragments in RLGS patterns by matching to the human genome sequence. A combination of RLGS and VGS was utilized to identify changes of chromosome 1–derived fragments in neuroblastoma. A NotI-EcoRV fragment was found to be absent frequently in neuroblastoma cell line RLGS patterns. Sequence prediction by VGS as well as cloning of the fragment showed that it contained a CpG island that is part of the human orthologue of the hamster homeobox gene Alx3. Expression analysis in a panel of human and mouse tissues showed predominant expression of ALX3 in brain tissue. Methylation-sensitive sequence analysis of the promoter region in neuroblastoma cell lines indicated that methylation of specific sequences correlated with repression of the ALX3 gene. Expression was re-induced after treatment with the methylation inhibitor 5-aza-2′-deoxycytidine. Promoter methylation analysis of ALX3 in primary neuroblastoma tumors, using methylation-sensitive polymerase chain reaction, found preferential ALX3 methylation in advanced-stage tumors. The VGS approach we have implemented in combination with RLGS is useful for the identification of genomic CpG island–related methylation changes or deletions in cancer. © 2002 Wiley-Liss, Inc. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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