This Article Full Text Full Text (PDF) Purchase Article View Shopping Cart 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 HighWire Citing Articles via Google Scholar Google Scholar Articles by Hoon, D. S. B. Articles by Seeger, R. C. Search for Related Content PubMed PubMed Citation Articles by Hoon, D. S. B. Articles by Seeger, R. C.

(American Journal of Pathology. 2001;159:493-500.)
© 2001 American Society for Investigative Pathology

Technical Advances

Ganglioside GM2/GD2 Synthetase mRNA Is a Marker for Detection of Infrequent Neuroblastoma Cells in Bone Marrow

Dave S. B. Hoon^*, Christine T. Kuo^*, Shenghua Wen, Hong Wang, Leonid Metelitsa, C. Patrick Reynolds and Robert C. Seeger

From the Department of Molecular Oncology,^*
John Wayne Cancer Institute, Santa Monica; the Division of Hematology-Oncology,
Department of Pediatrics, Children’s Hospital Los Angeles, University of Southern California School of Medicine Los Angeles; and the Children’s Cancer Group,
Arcadia, California

GalNAcß1-4(NeuAc2-3)Galß1-4Glcß1-Cer (GM2)/GalNAcß1-4(NeuAc2-8NeuAc2-3)Galß1-4Glcß1-1Cer (GD2) synthetase [ß-1,4-N-acetyl-galactosaminyl transferase (GalNAc-T)] mRNA, which encodes a key glycosyltransferase for ganglioside GD2 synthesis, was assessed as a molecular marker for detecting metastatic neuroblastoma cells in bone marrow (BM). GalNAc-T mRNA expression by neuroblastoma cell lines (n = 15), primary untreated neuroblastoma tumors (n = 29), morphologically normal BM (n = 22), peripheral blood stem cells (n = 10) from patients with cancers other than neuroblastoma, and blood mononuclear cells from normal donors (n = 17) was assessed by using reverse transcriptase-polymerase chain reaction (RT-PCR) and electrochemiluminescence detection assay (RT-PCR/ECL). BM harvested from 15 neuroblastoma patients was tested before and after ex vivo immunomagnetic bead purging, and results were compared to immunocytological analysis of the same specimens. All neuroblastoma cell lines (mean, 653 x 10³ ECL units) and primary tumors (mean, 683 x 10³ ECL units) were positive for significant expression of GalNAc-T mRNA compared to normal blood and BM cells. The RT-PCR/ECL assay could detect GalNAc-T mRNA in 100 pg of total RNA, and in a mixture of one neuroblastoma cell among 10⁷ normal BM or blood cells. Eight of 15 autologous BM cells harvested from patients with neuroblastoma had tumor cells detectable by immunocytology, and all 15 were positive for GalNAc-T mRNA. After ex vivo purging, none of the BM cells was immunocytology-positive, but six remained positive by the RT-PCR/ECL assay. GalNAc-T mRNA provides a specific and sensitive molecular marker for RT-PCR/ECL detection of infrequent neuroblastoma cells in BM.

This article has been cited by other articles:

				J. Stutterheim, A. Gerritsen, L. Zappeij-Kannegieter, I. Kleijn, R. Dee, L. Hooft, M. M. van Noesel, M. Bierings, F. Berthold, R. Versteeg, et al. PHOX2B Is a Novel and Specific Marker for Minimal Residual Disease Testing in Neuroblastoma J. Clin. Oncol., November 20, 2008; 26(33): 5443 - 5449. [Abstract] [Full Text] [PDF]

				T. Nakagawa, S. R. Martinez, Y. Goto, K. Koyanagi, M. Kitago, T. Shingai, D. A. Elashoff, X. Ye, F. R. Singer, A. E. Giuliano, et al. Detection of Circulating Tumor Cells in Early-Stage Breast Cancer Metastasis to Axillary Lymph Nodes Clin. Cancer Res., July 15, 2007; 13(14): 4105 - 4110. [Abstract] [Full Text] [PDF]

				K. Swerts, B. De Moerloose, C. Dhooge, J. Vandesompele, C. Hoyoux, K. Beiske, Y. Benoit, G. Laureys, and J. Philippe Potential Application of ELAVL4 Real-Time Quantitative Reverse Transcription-PCR for Detection of Disseminated Neuroblastoma Cells Clin. Chem., March 1, 2006; 52(3): 438 - 445. [Abstract] [Full Text] [PDF]

				I. Y. Cheung, M. S. Lo Piccolo, B. H. Kushner, and N.-K. V. Cheung Early Molecular Response of Marrow Disease to Biologic Therapy Is Highly Prognostic in Neuroblastoma J. Clin. Oncol., October 15, 2003; 21(20): 3853 - 3858. [Abstract] [Full Text] [PDF]

				C. T. Kuo, D. S.B. Hoon, H. Takeuchi, R. Turner, H.-J. Wang, D. L. Morton, and B. Taback Prediction of Disease Outcome in Melanoma Patients by Molecular Analysis of Paraffin-Embedded Sentinel Lymph Nodes J. Clin. Oncol., October 1, 2003; 21(19): 3566 - 3572. [Abstract] [Full Text] [PDF]

				G. Mehes, A. Luegmayr, R. Kornmuller, I. M. Ambros, R. Ladenstein, H. Gadner, and P. F. Ambros Detection of Disseminated Tumor Cells in Neuroblastoma: 3 Log Improvement in Sensitivity by Automatic Immunofluorescence plus FISH (AIPF) Analysis Compared with Classical Bone Marrow Cytology Am. J. Pathol., August 1, 2003; 163(2): 393 - 399. [Abstract] [Full Text] [PDF]

				I. Y. Cheung, M. S. Lo Piccolo, B. H. Kushner, K. Kramer, and N.-K. V. Cheung Quantitation of GD2 Synthase mRNA by Real-Time Reverse Transcriptase Polymerase Chain Reaction: Clinical Utility in Evaluating Adjuvant Therapy in Neuroblastoma J. Clin. Oncol., March 15, 2003; 21(6): 1087 - 1093. [Abstract] [Full Text] [PDF]