The importance of selecting the right internal control gene to study the effects of antenatal glucocorticoid administration in human placenta

Placenta. 2016 Aug:44:19-22. doi: 10.1016/j.placenta.2016.05.011. Epub 2016 May 27.

Abstract

RT-qPCR requires a suitable set of internal control genes (ICGs) for an accurate normalization. The usefulness of 7 previously published ICGs in the human placenta was analyzed according to the effects of betamethasone treatment, sex and fetal age. Raw RT-qPCR data of the ICGs were evaluated using published algorithms. The algorithms revealed that a reliable normalization was achieved using the geometrical mean of PPIA, RPL19, HMBS and SDHA. The use of a different subset ICGs out of the 7 investigated, although not statistically affected by the conditions, biased the results, as demonstrated through changes in expression of glucocorticoid receptor (NR3C1) mRNA as a target gene.

Keywords: B2M; Betamethasone; Co-regulation; Glucocorticoids; HMBS; HPRT1; Human placenta; Internal control gene; PPIA; RPL19; SDHA; YWHAZ; geNorm.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Electron Transport Complex II / genetics
  • Female
  • Gene Expression Profiling
  • Genes, Essential / genetics*
  • Glucocorticoids / pharmacology*
  • Glucocorticoids / therapeutic use
  • Humans
  • Hydroxymethylbilane Synthase / genetics
  • Peptidylprolyl Isomerase / genetics
  • Placenta / drug effects*
  • Placenta / metabolism
  • Pregnancy
  • Receptors, Glucocorticoid / genetics*
  • Ribosomal Proteins / genetics

Substances

  • Glucocorticoids
  • NR3C1 protein, human
  • RPL19 protein, human
  • Receptors, Glucocorticoid
  • Ribosomal Proteins
  • Electron Transport Complex II
  • SDHA protein, human
  • Hydroxymethylbilane Synthase
  • Peptidylprolyl Isomerase