N-myc gene copy number in neuroblastoma cell lines and resistance to experimental treatment

The N-myc oncogene is amplified in approximately 30% of neuroblastomas. It is well established that cases of neuroblastoma with amplified N-myc have markedly poorer prognosis than those in which N-myc copy number is not elevated. The mechanism for this association is not known but may be related to cellular resistance to radiation or cytotoxic drugs. Seven human neuroblastoma cell lines were used to investigate the relationship between N-myc copy number or expression and sensitivity to ionising radiation and to cisplatin. N-myc copy number was assessed by Southern blotting and hybridisation using the p-Nb1 probe. The signal produced by DNA from the cell lines was compared with that of single copy N-myc from normal human placental DNA. A range of N-myc copy numbers from 1 to 800 was found. Expression levels of N-myc mRNA were compared by "dot blotting" and subsequent hybridisation to the p-Nb1 probe. Radiosensitivity was assessed by surviving fraction at 2 Gy (SF2) following 60Co gamma irradiation. Values ranged from 0.13 to 0.52. Sensitivity to cisplatin was indicated by comparison of isoeffective concentrations (concentration required to produce 1 log cell kill). These ranged from 7.5 to 13 microM. Cisplatin studies showed a correlation between N-myc copy number (though not expression) and resistance to this drug. If this relationship is causal it may explain why treatment fails in those patients with an elevated N-myc copy number. However, no correlation was found between N-myc copy number or expression and sensitivity to radiation. It is possible that N-myc amplification confers resistance to some but not all treatments used in the therapy of neuroblastoma. Further investigations along these lines may lead to the identification of agents which are most appropriate for the treatment of neuroblastoma with amplified N-myc gene.