ArticleG3 (Bethesda, Md.)2014
Reference genes for expression studies in hypoxia and hyperglycemia models in human umbilical vein endothelial cells.
Article in G3 (Bethesda, Md.), 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed, 55 citations in OpenAlex.
- Putative Novel miR-1133 Is Associated with Oxidative Stress in Hyperglycemia-Induced Endothelial Cells.Biomedicines · 2026Article
- Endothelial Cells Differentiated from Human Induced Pluripotent Stem Cells Form Aligned Network Structures in Engineered Neural Tissue.Journal of functional biomaterials · 2025Article
- Selection of Reference Genes for Normalization of Gene Expression After Exposure of Human Endothelial and Epithelial Cells to Hypoxia.International journal of molecular sciences · 2025Article
- Transcriptomic analysis of iPSC-derived endothelium reveals adaptations to high altitude hypoxia in energy metabolism and inflammation.PLoS genetics · 2025Article
- Article
- Using RNA-seq to identify suitable housekeeping genes for hypoxia studies in human adipose-derived stem cells.BMC molecular and cell biology · 2023Article
- 3D In Vitro Blood-Brain-Barrier Model for Investigating Barrier Insults.Advanced science (Weinheim, Baden-Wurttemberg, Germany) · 2023Article
- In silico Identification of Hypoxic Signature followed by reverse transcription-quantitative PCR Validation in Cancer Cell LinesIranian biomedical journal · 2023Article
- A pleiotropic hypoxia-sensitiveScience advances · 2022Article
- Identification of appropriate housekeeping genes for gene expression studies in human renal cell carcinoma under hypoxic conditions.Molecular biology reports · 2022Article
- A reliable set of reference genes to normalize oxygen-dependent cytoglobin gene expression levels in melanoma.Scientific reports · 2021Article
- IRE1 Endoribonuclease Activity Modulates Hypoxic HIF-1α Signaling in Human Endothelial Cells.Biomolecules · 2020Article
- MicroRNA-29b-3p Promotes Human Retinal Microvascular Endothelial Cell Apoptosis via Blocking SIRT1 in Diabetic Retinopathy.Frontiers in physiology · 2019Article
- Sustained Intermittent Hypoxemia Induces Adiponectin Oligomers Redistribution and a Tissue-Specific Modulation of Adiponectin Receptor in Mice.Frontiers in physiology · 2019Article
- Effect of rosuvastatin on the expression of candidate geneExperimental and therapeutic medicine · 2018Article
- Selection of Reference Genes for Studies of Human Retinal Endothelial Cell Gene Expression by Reverse Transcription-Quantitative Real-Time Polymerase Chain Reaction.Gene reports · 2018Article
- Proangiogenic Effect of Metformin in Endothelial Cells Is via Upregulation of VEGFR1/2 and Their Signaling under Hyperglycemia-Hypoxia.International journal of molecular sciences · 2018Article
- Identification of suitable reference genes for real-time quantitative PCR analysis of hydrogen peroxide-treated human umbilical vein endothelial cells.BMC molecular biology · 2017Article
- Article
- Identification of Appropriate Housekeeping Genes for Gene Expression Analysis in Long-term Hypoxia-treated Kidney Cells.Advanced biomedical research · 2017Article
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Authors and funding
10 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Human umbilical vein endothelial cell (HUVEC)-based gene expression studies performed under hypoxia and/or hyperglycemia show huge potential for modeling endothelial cell response in cardiovascular disease and diabetes. However, such studies require reference genes that are stable across the whole range of experimental conditions. These reference genes have not been comprehensively defined to date. We applied human genome-wide microarrays and quantitative real-time PCR (qRT-PCR) on RNA obtained from primary HUVEC cultures that were incubated for 24 hr either in euglycemic or in hyperglycemic conditions and then subjected to short-term CoCl2-induced hypoxia for 1, 3, or 12 hr. Using whole-transcript arrays, we selected 10 commonly used reference genes with no significant expression variation across eight different conditions. These genes were ranked using NormFinder software according to their stability values. Consequently, five genes were selected for validation by qRT-PCR. These were ribosomal protein large P0 (RPLP0), transferrin receptor (TFRC), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-glucuronidase (GUSB), and β-actin (ACTB). All five genes displayed stable expression under hyperglycemia. However, only RPLP0 and TFRC genes were stable under hypoxia up to 12 hr. Under hyperglycemia combined with hypoxia up to 12 hr, the expression of RPLP0, TFRC, GUSB, and ACTB genes remained unchanged. Our findings strongly confirm that RPLP0 and TFRC are the most suitable reference genes for HUVEC gene expression experiments subjected to hypoxia and/or hyperglycemia for the given experimental conditions. We provide further evidence that even commonly known references genes require experimental validation for all conditions involved.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.