Background: In the last decade, microarray technology has been extensively used to evaluate gene expression profiles and genome imbalances. We have developed a microarray-based comparative genomic hybridization (CGH) approach to identify MYCN gene amplification and 1p36 chromosome loss, two markers of tumor aggressiveness in neuroblastoma.
Aim: The aim was to use microarray CGH technology to detect the two major prognostic markers for neuroblastoma, MYCN amplification and 1p36 chromosome deletion, in neuroblastoma patients and, therefore, confirm the usefulness of this approach in this cancer.
Methods: DNA was purified from 16 tumors containing at least 90% malignant neuroblasts and collected at the onset of disease. Pooled fluorescent-labeled reference and neuroblastoma tumor genomic DNA was hybridized to epoxide-coated glass slides on laboratory-made complementary DNA microarray. The microarray contained cDNA mapped at the 1p36.33–36.1 chromosomal region and MYCN gene. cDNA from the 2q33–q34 and 12p13 chromosomes was used as a control and Arabidopsis thaliana DNA was spotted to control unspecific hybridization. Fluorescence in situ hybridization analysis was also performed to validate results from the microarray CGH.
Results: Both MYCN amplification and 1p36 chromosome deletion were detected by microarray CGH. The sensitivity and specificity for 1p36 loss detection were 66.7% and 90.0%, respectively. The method had a sensitivity of 66.7% and specificity of 90.9% to detect MYCN amplification.
Discussion: Our results demonstrated that the microarray CGH can be efficiently applied to study DNA gain and loss of specific chromosome regions.
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This work was supported by the Fondazione Italiana per la Lotta al Neuroblastoma, Associazione Italiana per la Ricerca sul Cancro (AIRC), European Union 5th FP (QLRT-2001-02831 and QLRT-2001-02198) and the Ministero dell’Università e della Ricerca Scientifica e Tecnologica. We are grateful to the surgeons, clinicians, and pathologists of the Italian Cooperative Group for Neuroblastoma and to the Associazione Italiana di Ematologia e Oncologia Pediatrica for providing tumor samples.
Dr Scaruffi was partially supported by a Research Training Fellowship awarded by the International Agency for Research on Cancer. The authors have provided no information on conflicts of interest directly relevant to the content of this study.
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