RNA-Seq reveals spliceosome and proteasome genes as most consistent transcripts in human cancer cells

PLoS One. 2013 Sep 17;8(9):e72884. doi: 10.1371/journal.pone.0072884. eCollection 2013.

Abstract

Accurate quantification of gene expression by qRT-PCR relies on normalization against a consistently expressed control gene. However, control genes in common use often vary greatly between samples, especially in cancer. The advent of Next Generation Sequencing technology offers the possibility to better select control genes with the least cell to cell variability in steady state transcript levels. Here we analyze the transcriptomes of 55 leukemia samples to identify the most consistent genes. This list is enriched for components of the proteasome (ex. PSMA1) and spliceosome (ex. SF3B2), and also includes the translation initiation factor EIF4H, and many heterogeneous nuclear ribonucleoprotein genes (ex. HNRNPL). We have validated the consistency of our new control genes in 1933 cancer and normal tissues using publically available RNA-seq data, and their usefulness in qRT-PCR analysis is clearly demonstrated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Cell Survival / genetics
  • Cell Survival / physiology
  • Eukaryotic Initiation Factors / genetics
  • Eukaryotic Initiation Factors / metabolism
  • Gene Expression Regulation, Neoplastic / genetics
  • Gene Expression Regulation, Neoplastic / physiology
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Leukemia / genetics
  • Leukemia / metabolism
  • Proteasome Endopeptidase Complex / genetics
  • Proteasome Endopeptidase Complex / metabolism*
  • Spliceosomes / genetics
  • Spliceosomes / metabolism*

Substances

  • EIF4H protein, human
  • Eukaryotic Initiation Factors
  • Proteasome Endopeptidase Complex