gms | German Medical Science

62nd Annual Meeting of the German Society of Neurosurgery (DGNC)
Joint Meeting with the Polish Society of Neurosurgeons (PNCH)

German Society of Neurosurgery (DGNC)

7 - 11 May 2011, Hamburg

Selection of reference genes for quantitative real-time polymerase chain reaction in human meningiomas and arachnoidea

Meeting Abstract

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  • C. Pfister - Klinik für Neurochirurgie, Universitätsklinikum Tübingen
  • R. Ritz - Klinik für Neurochirurgie, Universitätsklinikum Tübingen
  • M. Tatagiba - Klinik für Neurochirurgie, Universitätsklinikum Tübingen
  • F. Roser - Klinik für Neurochirurgie, Universitätsklinikum Tübingen

Deutsche Gesellschaft für Neurochirurgie. Polnische Gesellschaft für Neurochirurgen. 62. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC), Joint Meeting mit der Polnischen Gesellschaft für Neurochirurgen (PNCH). Hamburg, 07.-11.05.2011. Düsseldorf: German Medical Science GMS Publishing House; 2011. DocMI.04.04

DOI: 10.3205/11dgnc202, URN: urn:nbn:de:0183-11dgnc2026

Published: April 28, 2011

© 2011 Pfister et al.
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Outline

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Objective: In meningiomas quantitative real-time reverse transcription-polymerase chain reaction (qPCR) is most frequently used for the accurate determination of gene expression using various reference genes. Although meningiomas are a heterogenous group of tissues, no data have been reported to validate reference genes for meningiomas and their control tissues.

Methods: At first 32 housekeeping genes were analyzed in meningiomas using geNorm, NormFinder and Bestkeeper-1 software. The six most suitable reference genes (EIF2B1, ELF1, PGK1, POLR2A, RPL30, RPL37A) plus GAPDH and ACTB were analyzed in 28 meningiomas, arachnoidea, dura and normal brain.

Results: geNorm and Bestkeeper-1 analysis identified RPL37A as the most stable expressed gene followed by RPL30 and PGK1, whereas NormFinder ranked RPL37A highest followed by PGK1 and EIF2B1. Commmonly used reference genes GAPDH and ACTB proofed to be unstable. The critical influence of reference genes on qPCR data analysis is shown for VEGFA transcription patterns, which indicates that a combination of RPL37A,RPL30 and PGK1 or RPL37A,PGK1 and EIF2B1 is most accurate.

Conclusions: RPL37A is the optimal reference gene for the normalization of gene expression in meningiomas, although the use of a combination of three reference genes would provide more stable results.