Gene Expression in Neuroblastoma

Abstract

The aim of this project is to identify potential target genes (NTG's) which may be regulated by in the cell line SK-N-SH which has no amplification of theoncogene N-myc. N-mjc target genes were isolated using a modification of the protocol developed to identify p53 binding sites; this was done by generating a NTG library by isolating genomic DNA from SK-N-SH cells, digesting to completion with Sau3A, and mixing with nuclear extract from the same cell line. N-myc antibody was added and DNA-protein complexes immunoprecipitated to isolate DNA with N-Myc bound. DNA recovered from these precipitates was then cloned into pBluescript SK+. 400 clones were isolated, 4 of which were partially sequenced and compared to sequence data bases. The NTG clones showed substantial sequence identity to a number of potentially interesting genes, among them several implicated in tumour suppression, and the expression of cell surface markers. Their significance for neuroblastoma is discussed.