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Identification of suitable reference genes for real-time qPCR in homocysteine-treated human umbilical vein endothelial cells

Author

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  • Xia Zhu
  • Lujun Zhang
  • Yangxi Hu
  • Jianliang Zhang

Abstract

The imbalance in homocysteine (Hcy) metabolism has been implicated in the pathogenesis of human diseases, including cardiovascular and neurodegenerative disorders. When attempting to identify gene expression profiles using quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR), the selection of suitable reference genes is important. Here, the expression levels of 10 commonly used reference genes were assessed for normalization of RT-qPCR in Hcy-treated human umbilical vein endothelial cells (HUVECs) and control cells. The suitability of eight selected candidate genes was comparatively analyzed across the tested samples and separately ranked by four programs, geNorm, NormFinder, BestKeeper, and the ΔCt method. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was the most stable gene in the final ranking using the RankAggreg package. Surprisingly, the β-actin (ACTB) levels decreased significantly in Hcy-treated HUVECs compared with control HUVECs (P

Suggested Citation

  • Xia Zhu & Lujun Zhang & Yangxi Hu & Jianliang Zhang, 2018. "Identification of suitable reference genes for real-time qPCR in homocysteine-treated human umbilical vein endothelial cells," PLOS ONE, Public Library of Science, vol. 13(12), pages 1-12, December.
    • Handle: RePEc:plo:pone00:0210087
    DOI: 10.1371/journal.pone.0210087
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