High nm23-H1/NDPK-A expression in Ewing tumors: paradoxical immunohistochemical reactivity and lack of prognostic significance

Int J Cancer. 1995 Apr 21;64(2):104-11. doi: 10.1002/ijc.2910640206.

Abstract

Expression of nm23-H1/NDPK-A has been reported to correlate inversely with metastasizing potential of rodent experimental cells and some human tumors. In the search for reliable molecular prognostic indicators for Ewing tumors (ET), a group of aggressive presumably neuroectodermal malignancies in children and adolescents, we studied nm23-H1/NDPK-A expression. Northern-blot and RT-PCR analyses were employed to semi-quantificatively measure nm23-H1 mRNA levels in ET cell lines and tissue extracts. A panel of monoclonal antibodies (MAbs) were used to evaluate protein abundance by Western blotting and immunohistochemistry. The nm23-H1/NDPK-A gene was also investigated on the DNA level to define possible genomic alterations. Our results revealed neither nm23-H1 allelic loss nor gene amplification and failed to show any significant variation in nm23-H1 mRNA or NDPK-A protein levels of primary or metastatic ET. NDPK-A protein levels were high and comparable to those of MCF-7 breast-cancer cells and to aggressive stage-IV neuroblastoma cell lines. nm23-H2/NDPK-B expression in ET was slightly more variable but generally lower than in MCF-7 cells. In the immunohistochemical analysis, however, discrepancies in the reactivity patterns with different antibodies were observed. Differential sensitivity to various fixation methods and heat treatment pointed to a structurally polymorphic NDPK-A protein. nm23-H1 expression studies using immunohistochemistry for prognostic counselling should thus be interpreted with caution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Antibodies, Monoclonal
  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • Blotting, Western
  • Child
  • Female
  • Flow Cytometry
  • Fluorescent Antibody Technique
  • Gene Expression Regulation, Enzymologic*
  • Gene Expression Regulation, Neoplastic*
  • Humans
  • Male
  • Molecular Sequence Data
  • Monomeric GTP-Binding Proteins*
  • NM23 Nucleoside Diphosphate Kinases
  • Nucleoside-Diphosphate Kinase / analysis*
  • Polymerase Chain Reaction / methods
  • Prognosis
  • RNA-Directed DNA Polymerase
  • Sarcoma, Ewing / chemistry*
  • Sarcoma, Ewing / enzymology
  • Sarcoma, Ewing / genetics
  • Sarcoma, Ewing / secondary
  • Transcription Factors / analysis*

Substances

  • Antibodies, Monoclonal
  • NM23 Nucleoside Diphosphate Kinases
  • Transcription Factors
  • NME1 protein, human
  • Nucleoside-Diphosphate Kinase
  • RNA-Directed DNA Polymerase
  • Monomeric GTP-Binding Proteins