title: Identification of suitable reference genes for real-time quantitative PCR analysis of hydrogen peroxide-treated human umbilical vein endothelial cells.
abstract: Oxidative stress can induce cell injury in vascular endothelial cells, which is the initial event in the development of atherosclerosis. Although quantitative real-time polymerase chain reaction (qRT-PCR) has been widely used in gene expression studies in oxidative stress injuries, using carefully validated reference genes has not received sufficient attention in related studies. The objective of this study, therefore, was to select a set of stably expressed reference genes for use in qRT-PCR normalization in oxidative stress injuries in human umbilical vein endothelial cells (HUVECs) induced by hydrogen peroxide (H2O2).
  
  
  
  Using geNorm analysis, we found that five stably expressed reference genes were sufficient for normalization in qRT-PCR analysis in HUVECs treated with H2O2. Genes with the most stable expression according to geNorm were U6, TFRC, RPLP0, GAPDH, and ACTB, and according to NormFinder were ALAS1, TFRC, U6, GAPDH, and ACTB.
  
  
  
  Taken together, our study demonstrated that the expression stability of reference genes may differ according to the statistical program used. U6, TFRC, RPLP0, GAPDH, and ACTB was the optimal set of reference genes for studies on gene expression performed by qRT-PCR assays in HUVECs under oxidative stress study.
authors.affiliation: Department of Cardiology, The Second Hospital of Jilin University, Changchun, 130041, Jilin, China.
authors.forename: Tianyi
authors.initials: T
authors.lastname: Li
authors.affiliation: Department of Cardiology, The Second Hospital of Jilin University, Changchun, 130041, Jilin, China.
authors.forename: Hongying
authors.initials: H
authors.lastname: Diao
authors.affiliation: Department of Cardiology, The Second Hospital of Jilin University, Changchun, 130041, Jilin, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhao
authors.affiliation: Department of Cardiology, The Second Hospital of Jilin University, Changchun, 130041, Jilin, China.
authors.forename: Yue
authors.initials: Y
authors.lastname: Xing
authors.affiliation: Department of Cardiology, The Second Hospital of Jilin University, Changchun, 130041, Jilin, China.
authors.forename: Jichang
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Cardiology, The Second Hospital of Jilin University, Changchun, 130041, Jilin, China.
authors.forename: Ning
authors.initials: N
authors.lastname: Liu
authors.affiliation: Department of Cardiology, The Second Hospital of Jilin University, Changchun, 130041, Jilin, China.
authors.forename: Youyou
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Department of Cardiology, The Second Hospital of Jilin University, Changchun, 130041, Jilin, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Tian
authors.affiliation: Department of Cardiology, The Second Hospital of Jilin University, Changchun, 130041, Jilin, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Sun
authors.affiliation: Department of Cardiology, The Second Hospital of Jilin University, Changchun, 130041, Jilin, China. liubin3333@vip.sina.com.
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
mesh_terms: D042783:Endothelial Cells / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism*; D018384:Oxidative Stress; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D059467:Transcriptome
chemical_list: D006861:Hydrogen Peroxide
keywords: Human umbilical vein endothelial cells; Hydrogen peraoxide; Normalization; Reference genes; qRT-PCR