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

Autor:	Borja Fernández, Juan Robledo-Carmona, Carmen Rueda-Martínez, Gemma Sánchez-Espín, Manuel F. Jiménez-Navarro, Maria Mataró, Miguel Such-Martínez, Oscar Lamas
Přispěvatelé:	[Rueda-Martínez,C, Lamas,O, Mataró,MJ, Robledo-Carmona,J, Sánchez-Espín,G, Jiménez-Navarro,M, Such-Martínez,M] UGC del Corazón, Instituto de Investigación Biomédica de Málaga (IBIMA), Hospital Clínico Universitario Virgen de la Victoria, Málaga, Spain. [Jiménez-Navarro,M] Departamento de Medicina, Facultad de Medicina, Universidad de Málaga, Spain. [Fernández, B] Departamento de Biología Animal, Instituto de Investigación Biomédica de Málaga (IBIMA), Facultad de Ciencias, Universidad de Málaga, Spain., This work was supported in part by grants PI-0689/2010 (Consejería de Salud, Junta de Andalucía, Spain), P10-CTS-06068 (Consejería de Innovación, Ciencia y Empresa, Junta de Andalucía, Spain), and Red de Investigación Cardiovascular (RIC) from Redes Temáticas de Investigación Cooperativa (RETIC)
Rok vydání:	2014
Předmět:	Male Aortic valve Pathology Gene Identification and Analysis Myocardial Infarction Chemicals and Drugs::Nucleic Acids Nucleotides and Nucleosides::Nucleic Acids::DNA::DNA Single-Stranded::DNA Complementary [Medical Subject Headings] Gene Expression lcsh:Medicine Vascular Medicine Phenomena and Processes::Genetic Phenomena::Genetic Processes::Gene Expression [Medical Subject Headings] Cohort Studies Anatomy::Cardiovascular System::Blood Vessels::Arteries::Aorta [Medical Subject Headings] Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::Humans [Medical Subject Headings] Bicuspid aortic valve Reference genes Gene expression Morphogenesis Medicine and Health Sciences lcsh:Science Aorta Anatomy::Cardiovascular System::Heart::Heart Valves::Aortic Valve [Medical Subject Headings] Multidisciplinary Algoritmos Nuclear Proteins RNA-Binding Proteins Middle Aged Congenital Heart Defects Neoplasm Proteins Humanos medicine.anatomical_structure Cardiovascular Diseases Research Design Diseases::Pathological Conditions Signs and Symptoms::Pathological Conditions Anatomical::Dilatation Pathologic [Medical Subject Headings] cardiovascular system Female RNA extraction Anatomy Algorithms Cyclin-Dependent Kinase Inhibitor p27 Dilatation Pathologic Research Article Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Nucleic Acid Denaturation [Medical Subject Headings] Dilatación Patológica medicine.medical_specialty Expresión Génica Clinical Research Design Health Care::Population Characteristics::Demography [Medical Subject Headings] Cardiology Aortic Diseases Desnaturalización de Ácido Nucleico Diseases::Cardiovascular Diseases::Vascular Diseases::Aortic Diseases::Aortic Aneurysm [Medical Subject Headings] In Vitro Techniques Biology Real-Time Polymerase Chain Reaction Research and Analysis Methods Molecular Genetics Diseases::Cardiovascular Diseases::Heart Diseases::Heart Valve Diseases [Medical Subject Headings] Anatomy::Tissues [Medical Subject Headings] medicine.artery Valvular Diseases Enfermedades de las Válvulas Cardíacas Aneurisma de la Aorta Ascending aorta Genetics medicine Humans Demografía Birth Defects ADN Complementario Gene Aged Congenital Hereditary Myopathies Clinical Genetics Acute Cardiovascular Problems lcsh:R 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] Biology and Life Sciences Analytical Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Nucleic Acid Amplification Techniques::Polymerase Chain Reaction [Medical Subject Headings] Human Genetics medicine.disease Reacción en Cadena en Tiempo Real de la Polimerasa Válvula Aórtica Cardiovascular Anatomy Reacción en Cadena de la Polimerasa lcsh:Q Information Science::Information Science::Computing Methodologies::Algorithms [Medical Subject Headings] Developmental Biology
Zdroj:	PLoS ONE, Vol 9, Iss 5, p e97449 (2014) PLoS ONE
ISSN:	1932-6203
Popis:	Journal Article; 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. Yes
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2c3a8f5ad173faed66077c1b64c68c32 https://doi.org/10.1371/journal.pone.0097449 Zobrazit plný text záznamu