MDM2 gene amplification and lack of p53 point mutations in Hodgkin and Reed-Sternberg cells: results from single-cell polymerase chain reaction and molecular cytogenetic studies

Hodkin's disease (HD) is the most common haematological malignancy after ch ronic lymphocytic leukaemia, but very little is known about its pathogenesi s or the genetic events that contribute to the malignant phenotype of the t umour cells, p53 is assumed to play an important role in the pathogenesis o f HD, based on the observation that p53 protein is frequently accumulated i n Hodgkin and Reed-Sternberg (H & RS) cells, We investigated single H & RS cells from five different HD patients for point mutations at the genomic le vel using multiplex polymerase chain reaction amplification and subsequent sequencing. No point mutations were detected in 50 single H & RS cells anal ysed. Hence, accumulation of p53 protein cannot be explained by mutations w ithin the gene. A genome-wide screening for genomic imbalances using compar ative genomic hybridization revealed gain on chromosome 12q14, i.e. the map ping position of the MDM2 gene in several HD cases. Therefore, we assessed the copy number of the MDM2 gene using fluorescence in situ hybridization, In four out of six HD cases analysed, the copy number of the MDM2 gene was found to be increased. As gene amplification is frequently associated with protein overexpression, the observed accumulation of p53 in the nuclei of H & RS cells could be as a result of elevated MDM2 protein Levels resulting in stabilization of p53 protein.