Neuron Glia Biology



Transcriptional profiling in an MPNST-derived cell line and normal human Schwann cells


PHILIP R. LEE a1, JONATHAN E. COHEN a1, ELISABETTA A. TENDI a1, ROBERT FARRER a2, GEORGE H. DE VRIES a2, KEVIN G. BECKER a3 and R. DOUGLAS FIELDS a1c1
a1 Section on Nervous System Development and Plasticity, NICHD, National Institutes of Health, Bethesda, MD
a2 Department of Cell Biology, Neurobiology and Anatomy, Loyola University of Chicago Stritch School of Medicine, Chicago, IL
a3 Gene Expression and Genomics Unit, NIA, National Institutes of Health, Baltimore, MD

Article author query
lee pr   [PubMed][Google Scholar] 
cohen je   [PubMed][Google Scholar] 
tendi ea   [PubMed][Google Scholar] 
farrer r   [PubMed][Google Scholar] 
de vries gh   [PubMed][Google Scholar] 
becker kg   [PubMed][Google Scholar] 
fields rd   [PubMed][Google Scholar] 

Abstract

cDNA microarrays were utilized to identify abnormally expressed genes in a malignant peripheral nerve sheath tumor (MPNST)-derived cell line, T265, by comparing the mRNA abundance profiles with that of normal human Schwann cells (nhSCs). The findings characterize the molecular phenotype of this important cell-line model of MPNSTs, and elucidate the contribution of Schwann cells in MPNSTs. In total, 4608 cDNA sequences were screened and hybridizations replicated on custom cDNA microarrays. In order to verify the microarray data, a large selection of differentially expressed mRNA transcripts were subjected to semi-quantitative reverse transcription PCR (LightCycler). Western blotting was performed to investigate a selection of genes and signal transduction pathways, as a further validation of the microarray data. The data generated from multiple microarray screens, semi-quantitative RT–PCR and Western blotting are in broad agreement. This study represents a comprehensive gene-expression analysis of an MPNST-derived cell line and the first comprehensive global mRNA profile of nhSCs in culture. This study has identified [similar]900 genes that are expressed abnormally in the T265 cell line and detected many genes not previously reported to be expressed in nhSCs. The results provide crucial information on the T265 cells that is essential for investigation using this cell line in experimental studies in neurofibromatosis type I (NF1), and important information on normal human Schwann cells that is applicable to a wide range of studies on Schwann cells in cell culture.


Key Words: Cancer; NF1; MPNST; T265; Schwann cell; cDNA microarray.

Correspondence:
c1 R. Douglas Fields, Section on Nervous System Development and Plasticity, Bldg. 35, Rm. 2A211, MSC 3713 NICHD, National Institutes of Health, Bethesda, MD 20892, USA. phone: +1 301 480 3209; fax: +1 301 496 9630. email: Fieldsd@mail.nih.gov