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.

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.

• Authors: Orsolya Giricz, Janelle L Lauer-Fields, Gregg B Fields
• Journal: Analytical biochemistry (Anal Biochem) Vol. 380 Issue 1 Pg. 137-9 (Sep 01 2008) ISSN: 1096-0309 [Electronic] United States
• PMID: 18554498 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• RNA, Messenger
• RNA, Ribosomal, 18S
• Glyceraldehyde-3-Phosphate Dehydrogenases

Topics

• Cell Line, Tumor
• Gene Expression Profiling (standards)
• Gene Expression Regulation, Neoplastic
• Glyceraldehyde-3-Phosphate Dehydrogenases (genetics)
• Humans
• Melanoma (genetics)
• Polymerase Chain Reaction (methods)
• RNA, Messenger (genetics)
• RNA, Ribosomal, 18S (genetics)
• Reference Standards
• Time Factors