This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Gazitt, Y. Articles by Graham-Pole, J. Search for Related Content PubMed PubMed Citation Articles by Gazitt, Y. Articles by Graham-Pole, J.

Expression of N-myc, c-myc, and MDR-1 Proteins in Newly Established Neuroblastoma Cell Lines: A Study by Immunofluorescence Staining and Flow Cytometry¹

Department of Pediatrics, Division of Hematology/Oncology, University of Florida Medical Center, Gainesville, Florida 32610

A methodology for rapid isolation of neuroblastoma cells from marrow with metastatic neuroblastoma cells was developed using a cocktail of five antibodies and magnetic microspheres coated with secondary antibodies. Cells bound to microspheres were released by brief exposure to chymopapain, followed by repeated culture of released cells in serum-supplemented Dulbecco's modified Eagle's medium and selection for adherent cells. Using this methodology, over 35 primary cell lines were obtained free of contaminating normal cells. Detailed analyses of over 14 cell lines revealed gross differences in cell phenotype, size, morphology development of neurite processes, and doubling time (40 to 80 h). All cell lines expressed the M_r 145,000 neurofilament, and a few expressed the M_r 200,000 neurofilament, with very little or no expression of the M_r 68,000 neurofilament. Eighty % of all cells lines had near-diploid DNA content. High expression of the MDR-1 protein was detected in six of the 22 cell lines tested.

Great heterogeneity was observed in the expression of N-myc oncoprotein, with ten of 13 patients overexpressing the protein. c-myc oncoprotein was also expressed in all cell lines; however, the level of expression was 4- to 10-fold lower than the N-myc oncoprotein. Localization studies of c-myc and N-myc oncoproteins on the level of light microscopy and electron microscopy revealed exclusive nuclear localization of c-myc, whereas N-myc was localized to the nucleus and to the cytoplasm.

¹ This work was supported partly by a grant from "STOP! Children's Cancer, Inc." and partly by the Division of Sponsored Research of the University of Florida (Y. G.).

² To whom requests for reprints should be addressed.

Received 8/19/91. Accepted 2/26/92.