Identification and validation of reference genes for quantitative RT-PCR analysis of retinal pigment epithelium cells under hypoxia and/or hyperglycemia

Gene
Xin LiuGuanfang Su

Abstract

Retinal pigment epithelium (RPE) cell-based gene expression studies performed under hypoxia and/or hyperglycemia show huge potential for modeling cell responses in diabetic retinopathy, retinopathy of prematurity and other retinal diseases. However, normalization of gene expression on RPE cells under those conditions has commonly been done using either GAPDH or β-actin as reference genes without any validation of their expression stability. Therefore, we aimed to establish a suitable set of reference genes for studies on RPE cells cultured under both normal culturing glucose and atmospheric oxygen tension (normoxia, 21%), under a low oxygen tension (hypoxia, 1%), under a high glucose growth medium (25 mmol/l) and under the combination of the two changed conditions above for distinct time points taking together from 24h to 7 days. Quantitative real-time PCR (qRT-PCR) was applied on RNA obtained from a cell line, ARPE-19. Stability of 14 commonly used reference genes was assessed and ranked according to their stability values using the geNorm and NormFinder softwares with the aim to find the most stable expressed gene under all conditions. Our findings confirm that HPRT1, GUSB and PPIA are the most suitable reference genes for RP...Continue Reading

References

Feb 25, 2003·Progress in Retinal and Eye Research·A N WitmerR O Schlingemann
Apr 16, 2003·Archives of Ophthalmology·Norbert D Wangsa-Wirawan, Robert A Linsenmeier
Sep 24, 2004·Experimental Eye Research·Jeffrey W SallSteven E Katz
Jul 1, 2005·Physiological Reviews·Olaf Strauss
Sep 24, 2005·Transplantation·Sílvia Rodriguez-Mulero, Eduard Montanya
Jun 15, 2007·The British Journal of Ophthalmology·Farzin ForooghianLee Timms
Jun 29, 2007·BMC Molecular Biology·Harun M SaidDirk Vordermark
Jan 24, 2009·Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie·Peng ZhangJian Zhou
Oct 13, 2009·BMC Molecular Biology·Casper Bindzus FoldagerMartin Lind
Mar 11, 2010·Methods : a Companion to Methods in Enzymology·Sean TaylorMarie Nguyen
Sep 7, 2014·G3 : Genes - Genomes - Genetics·Sherin BakhashabJolanta U Weaver
May 6, 2015·Progress in Retinal and Eye Research·Rafael Simó, Cristina Hernández

❮ Previous
Next ❯

Citations

Jul 1, 2017·Journal of Agricultural and Food Chemistry·Katja FredeSusanne Baldermann
Jul 15, 2017·Journal of Diabetes Research·Qiaoyun GongGuanfang Su
Sep 24, 2019·Frontiers in Neuroscience·Teresa TsaiStephanie C Joachim
Aug 28, 2020·International Journal of Molecular Sciences·Abdullah Al-AniMark Ungrin
Jan 27, 2020·Biomedicine & Pharmacotherapy = Biomédecine & Pharmacothérapie·Irene Bissoli, Claudio Muscari

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.