Identification of valid reference housekeeping genes for gene expression analysis in tumor neovascularization studies

Monica Rienzo, Concetta Schiano, Amelia Casamassimi, Vincenzo Grimaldi, Teresa Infante, Claudio Napoli

Research output: Contribution to journalArticle

Abstract

Introduction: Real time RT-PCR is a widely used technique to evaluate and confirm gene expression data obtained in different cell systems and experimental conditions. However, there are many conflicting reports about the same gene or sets of gene expression. A common method is to report the interest gene expression relative to an internal control, usually a housekeeping gene (HKG), which should be constant in cells independently of experimental conditions. Materials and Methods: In this study, the expression stability of ten HKGs was considered in parallel in two cell systems (endothelial and osteosarcoma cells): beta actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), TATA box binding protein (TBP), hypoxanthine phosphoribosyl-transferase 1 (HPRT1), Cyclophilin A (PPIA), beta-2-microglobulin (B2M), glucuronidase beta (GUSB), eukaryotic translation elongation factor 1 alpha1 (EEF1A1), transferrin receptor (TFRC), ribosomal protein S18 (RPS18). In order to study the stability of candidate reference genes, data have been also analyzed by several algorithms (geNorm, NormFinder, BestKeeper and delta-Ct method). Results and Conclusions: The overall analysis obtained by the comprehensive ranking showed that RPS18 and PPIA are appropriate internal reference genes for tumor neovascularization studies where it is necessary to analyze both systems at the same time.

Original languageEnglish
Pages (from-to)211-218
Number of pages8
JournalClinical and Translational Oncology
Volume15
Issue number3
DOIs
Publication statusPublished - 2013

Keywords

  • Endothelial cells
  • Housekeeping gene
  • Neovascularization
  • Osteosarcoma cells
  • Real time RT-PCR

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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