Gene amplification in PNETs/medulloblastomas: mapping of a novel amplified gene within the MYCN amplicon
- Michael C Frühwalda,b,c,
- M Sue O'Dorisio*,a,
- Laura J Rushb,d,
- Jill L Reitere,
- Dominic J Smiragliab,
- Gail Wengerf,
- Joseph F Costellog,
- Peter S Whiteh,
- Ralf Kraheb,
- Garrett M Brodeurh,
- Christoph Plassb
- aDivision of Pediatric Hematology and Oncology, The Ohio State University and the Comprehensive Cancer Center, Columbus, Ohio 43210, USA, bDivision of Human Cancer Genetics, The Ohio State University and the Comprehensive Cancer Center, Columbus, Ohio 43210, USA, cDepartment of Neuroscience, The Ohio State University and the Comprehensive Cancer Center, Columbus, Ohio 43210, USA, dDepartment of Veterinary Biosciences, The Ohio State University and the Comprehensive Cancer Center, Columbus, Ohio 43210, USA, eDepartment of Biochemistry and Molecular Biology, Mayo Clinic, Rochester, Minnesota 55905, USA, fDepartment of Pathology, The Ohio State University and the Comprehensive Cancer Center, Columbus, Ohio 43210, USA, gUniversity of California, San Francisco, CA 94115-0128, USA, hDivision of Oncology, The Children's Hospital of Philadelphia and the University of Pennsylvania, Philadelphia, Pennsylvania 19104-4318, USA
- Dr Frühwald, Westfälische Wilhelms-Universität Münster, Klinik und Poliklinik für Kinderheilkunde, Pädiatrische Hämatologie/Onkologie, Albert-Schweizer-Strasse 33, 48149 Münster, Germany,fruhwald{at}uni-muenster.de or Dr Plass,plass-1{at}medctr.osu.edu
- Revised 14 February 2000
- Accepted 18 February 2000
Abstract
OBJECTIVES The pathological entity of primitive neuroectodermal tumour/medulloblastoma (PNET/MB) comprises a very heterogeneous group of neoplasms on a clinical as well as on a molecular level. We evaluated the importance of DNA amplification in medulloblastomas and other primitive neuroectodermal tumours (PNETs) of the CNS.
METHOD Restriction landmark genomic scanning (RLGS), a method that allows the detection of low level amplification, was used. RLGS provides direct access to DNA sequences circumventing positional cloning efforts. Furthermore, we analysed several samples by CGH.
DESIGN Twenty primary medulloblastomas, five supratentorial PNETs, and five medulloblastoma cell lines were studied.
RESULTS Although our analysis confirms that gene amplification is generally a rare event in childhood PNET/MB, we found a total of 17 DNA fragments that were amplified in seven different tumours. Cloning and sequencing of several of these fragments confirmed the previous finding ofMYC amplification in the cell line D341 Med and identified novel DNA sequences amplified in PNET/MB. We describe for the first time amplification of the novel gene,NAG, in a subset of PNET/MB. Despite genomic amplification, NAG was not overexpressed in the tumours studied. We have determined thatNAG maps less than 50 kb 5′ ofDDX1 and approximately 400 kb telomeric ofMYCN on chromosome 2p24.
CONCLUSION We found a similar but slightly higher frequency of amplification than previously reported. We present several DNA fragments that may belong to the CpG islands of novel genes amplified in a small subset of PNET/MB. As an example we describe for the first time the amplification ofNAG in the MYCNamplicon in PNET/MB.
Footnotes
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↵* Present address: Department of Pediatric Hematology and Oncology, The University of Iowa, Iowa City, Iowa 52242, USA
