Identification of suitable reference genes for real-time quantitative PCR analysis of hydrogen peroxide-treated human umbilical vein endothelial cells

BMC Molecular Biology

Research Article

Xin Tian¹  Tianyi Li¹  Youyou Yan¹  Wei Sun¹  Hongying Diao¹  Lei Zhao¹  Jichang Zhang¹  Bin Liu¹  Yue Xing¹  Ning Liu¹ 

[1] Department of Cardiology, The Second Hospital of Jilin University, 130041, Changchun, Jilin, China;

关键词: Hydrogen peraoxide;    Human umbilical vein endothelial cells;    qRT-PCR;    Reference genes;    Normalization;   

DOI  :  10.1186/s12867-017-0086-z

 received in 2016-06-08, accepted in 2017-03-21,  发布年份 2017

来源: Springer

【 摘 要 】

BackgroundOxidative 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).ResultsUsing 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.ConclusionTaken 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.

【 授权许可】

CC BY   
© The Author(s) 2017

【 图 表 】

Fig. 7