Validierung von Microarray-Genexpressionsdaten bei Neuroblastomen mittels quantitativer Real-Time PCR

Neuroblastoma is the most common extracranial solid tumor of childhood under five years of age. It is a neuroblastic tumor of the primordial neural crest demonstrating diverse clinical and biological characteristics and behaviour. Several genetic features have been identified that are characteristics of neuroblastomas. Amplification of the MYCN proto-oncogene is associated with advanced tumor stage, progressive disease and correlates with a greatly increased risk of fatal outcome. MYCN amplified tumors and non amplified tumors were analyzed by quantitative real-time PCR to validate microarray expression profiles of five differentially expressed genes. Quantitative real-time PCR confirmed the change in expression of four of five genes identified by microarray analysis. To determine whether upregulated genes, Auroa-A and Trap1 were regulated by N-Myc the expression of these genes were analyzed in a cell line that carries a tetracycline regulatable allele of N-Myc. Both, Aurora-A and Trap1 were induced in response to induction of MYCN expression. Finally, neuroblastoma cell lines and neuroblastoma tumors were analyzed by Taqman PCR to detect Aurora-A amplification. In conclusion, amplification of Aurora-A has not been detected in our experiments.