Publication:
Selection of Reference Genes for Quantitative Real Time PCR (qPCR) Assays in Tissue from Human Ascending Aorta.

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Date

2014-05-19

Authors

Rueda-Martínez, Carmen
Lamas, Óscar
Mataró, María José
Robledo-Carmona, Juan
Sánchez-Espín, Gemma
Jiménez-Navarro, Manuel
Such-Martínez, Miguel
Fernández, Borja

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Public Library of Science
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Abstract

Dilatation of the ascending aorta (AAD) is a prevalent aortopathy that occurs frequently associated with bicuspid aortic valve (BAV), the most common human congenital cardiac malformation. The molecular mechanisms leading to AAD associated with BAV are still poorly understood. The search for differentially expressed genes in diseased tissue by quantitative real-time PCR (qPCR) is an invaluable tool to fill this gap. However, studies dedicated to identify reference genes necessary for normalization of mRNA expression in aortic tissue are scarce. In this report, we evaluate the qPCR expression of six candidate reference genes in tissue from the ascending aorta of 52 patients with a variety of clinical and demographic characteristics, normal and dilated aortas, and different morphologies of the aortic valve (normal aorta and normal valve n = 30; dilated aorta and normal valve n = 10; normal aorta and BAV n = 4; dilated aorta and BAV n = 8). The expression stability of the candidate reference genes was determined with three statistical algorithms, GeNorm, NormFinder and Bestkeeper. The expression analyses showed that the most stable genes for the three algorithms employed were CDKN1β, POLR2A and CASC3, independently of the structure of the aorta and the valve morphology. In conclusion, we propose the use of these three genes as reference genes for mRNA expression analysis in human ascending aorta. However, we suggest searching for specific reference genes when conducting qPCR experiments with new cohort of samples.

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Journal Article;

MeSH Terms

Medical Subject Headings::Diseases::Cardiovascular Diseases::Heart Diseases::Heart Valve Diseases
Medical Subject Headings::Anatomy::Cardiovascular System::Heart::Heart Valves::Aortic Valve
Medical Subject Headings::Information Science::Information Science::Computing Methodologies::Algorithms
Medical Subject Headings::Anatomy::Cardiovascular System::Blood Vessels::Arteries::Aorta
Medical Subject Headings::Diseases::Cardiovascular Diseases::Vascular Diseases::Aortic Diseases::Aortic Aneurysm
Medical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Nucleic Acid Amplification Techniques::Polymerase Chain Reaction::Real-Time Polymerase Chain Reaction
Medical Subject Headings::Health Care::Population Characteristics::Demography
Medical Subject Headings::Phenomena and Processes::Genetic Phenomena::Genetic Processes::Gene Expression
Medical Subject Headings::Diseases::Pathological Conditions, Signs and Symptoms::Pathological Conditions, Anatomical::Dilatation, Pathologic
Medical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Nucleic Acid Amplification Techniques::Polymerase Chain Reaction
Medical Subject Headings::Chemicals and Drugs::Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids::DNA::DNA, Single-Stranded::DNA, Complementary
Medical Subject Headings::Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Nucleic Acid Denaturation
Medical Subject Headings::Anatomy::Tissues
Medical Subject Headings::Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans

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Keywords

Enfermedades de las Válvulas Cardíacas, Válvula Aórtica, Algoritmos, Aneurisma de la Aorta, Reacción en Cadena en Tiempo Real de la Polimerasa, Demografía, Expresión Génica, Dilatación Patológica, Reacción en Cadena de la Polimerasa, ADN Complementario, Desnaturalización de Ácido Nucleico, Humanos

Citation

Rueda-Martínez C, Lamas O, Mataró MJ, Robledo-Carmona J, Sánchez-Espín G, Jiménez-Navarro M, et al. Selection of Reference Genes for Quantitative Real Time PCR (qPCR) Assays in Tissue from Human Ascending Aorta. PLoS ONE. 2014; 9(5):e97449