The normalization of gene expression data in melanoma

Investigating the use of glyceraldehyde 3-phosphate dehydrogenase and 18S ribosomal RNA as internal reference genes for quantitative real-time PCR

Orsolya Giricz, Janelle L. Lauer-Fields, Gregg B. Fields

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

We examined a panel of 26 melanoma and fibroblast samples (tissues and cultured cells) to evaluate the suitability of two commonly used housekeeping genes, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and 18S ribosomal RNA (rRNA), for quantitative real-time PCR. Both genes showed significant variations within the individual cell line and tissue groups. Although no overall trends were observed in the expression of the 18S rRNA, GAPDH was up-regulated in melanoma tissue and cultured cells compared with the corresponding normal samples. In melanoma and fibroblast cell lines and tissues, absolute quantification appears to be more appropriate than normalizing messenger RNA (mRNA) expression via GAPDH or 18S rRNA housekeeping genes.

Original languageEnglish (US)
Pages (from-to)137-139
Number of pages3
JournalAnalytical Biochemistry
Volume380
Issue number1
DOIs
StatePublished - Sep 1 2008
Externally publishedYes

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Glyceraldehyde-3-Phosphate Dehydrogenases
Gene expression
Real-Time Polymerase Chain Reaction
Melanoma
Genes
18S Ribosomal RNA
Cells
Tissue
Gene Expression
Essential Genes
Fibroblasts
Cultured Cells
Cell Line
rRNA Genes

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

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abstract = "We examined a panel of 26 melanoma and fibroblast samples (tissues and cultured cells) to evaluate the suitability of two commonly used housekeeping genes, glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and 18S ribosomal RNA (rRNA), for quantitative real-time PCR. Both genes showed significant variations within the individual cell line and tissue groups. Although no overall trends were observed in the expression of the 18S rRNA, GAPDH was up-regulated in melanoma tissue and cultured cells compared with the corresponding normal samples. In melanoma and fibroblast cell lines and tissues, absolute quantification appears to be more appropriate than normalizing messenger RNA (mRNA) expression via GAPDH or 18S rRNA housekeeping genes.",
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