naa a22 4500 510763707 CHVBK 20180411083148.0 cr unu---uuuuu 180411e20130301xx s 000 0 eng 10.1007/s12094-012-0904-1 doi (NATIONALLICENCE)springer-10.1007/s12094-012-0904-1 Identification of valid reference housekeeping genes for gene expression analysis in tumor neovascularization studies [Elektronische Daten] [Monica Rienzo, Concetta Schiano, Amelia Casamassimi, Vincenzo Grimaldi, Teresa Infante, Claudio Napoli] Introduction: Real time RT-PCR is a widely used technique to evaluate and confirm gene expression data obtained in different cell systems and experimental conditions. However, there are many conflicting reports about the same gene or sets of gene expression. A common method is to report the interest gene expression relative to an internal control, usually a housekeeping gene (HKG), which should be constant in cells independently of experimental conditions. Materials and Methods: In this study, the expression stability of ten HKGs was considered in parallel in two cell systems (endothelial and osteosarcoma cells): beta actin (ACTB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), TATA box binding protein (TBP), hypoxanthine phosphoribosyl-transferase 1 (HPRT1), Cyclophilin A (PPIA), beta-2-microglobulin (B2M), glucuronidase beta (GUSB), eukaryotic translation elongation factor 1 alpha1 (EEF1A1), transferrin receptor (TFRC), ribosomal protein S18 (RPS18). In order to study the stability of candidate reference genes, data have been also analyzed by several algorithms (geNorm, NormFinder, BestKeeper and delta-Ct method). Results and Conclusions: The overall analysis obtained by the comprehensive ranking showed that RPS18 and PPIA are appropriate internal reference genes for tumor neovascularization studies where it is necessary to analyze both systems at the same time. Federación de Sociedades Españolas de Oncología (FESEO), 2012 Endothelial cells nationallicence Housekeeping gene nationallicence Neovascularization nationallicence Osteosarcoma cells nationallicence Real time RT-PCR nationallicence Rienzo Monica Department of General Pathology, Second University of Naples, Via L. De Crecchio 7, 80138, Naples, Italy aut Schiano Concetta Department of General Pathology, Second University of Naples, Via L. De Crecchio 7, 80138, Naples, Italy aut Casamassimi Amelia Department of General Pathology, Second University of Naples, Via L. De Crecchio 7, 80138, Naples, Italy aut Grimaldi Vincenzo Department of General Pathology, Chair of Clinical Pathology, Excellence Research Centre on Cardiovascular Disease, U.O.C. Immunohematology, Transfusion Medicine and Transplant Immunology (SIMT), Regional Reference Laboratory of Transplant Immunology (LIT), Azienda Universitaria Policlinico (AOU), 1st School of Medicine, Second University of Naples, Piazza Miraglia 2, 80138, Naples, Italy aut Infante Teresa IRCCS SDN Foundation, Via Gianturco 113, 80134, Naples, Italy aut Napoli Claudio Department of General Pathology, Chair of Clinical Pathology, Excellence Research Centre on Cardiovascular Disease, U.O.C. Immunohematology, Transfusion Medicine and Transplant Immunology (SIMT), Regional Reference Laboratory of Transplant Immunology (LIT), Azienda Universitaria Policlinico (AOU), 1st School of Medicine, Second University of Naples, Piazza Miraglia 2, 80138, Naples, Italy aut Clinical and Translational Oncology Springer Milan 15/3(2013-03-01), 211-218 1699-048X 15:3<211 2013 15 12094 https://doi.org/10.1007/s12094-012-0904-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12094-012-0904-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Rienzo Monica Department of General Pathology, Second University of Naples, Via L. De Crecchio 7, 80138, Naples, Italy aut NATIONALLICENCE 700 1- Schiano Concetta Department of General Pathology, Second University of Naples, Via L. De Crecchio 7, 80138, Naples, Italy aut NATIONALLICENCE 700 1- Casamassimi Amelia Department of General Pathology, Second University of Naples, Via L. De Crecchio 7, 80138, Naples, Italy aut NATIONALLICENCE 700 1- Grimaldi Vincenzo Department of General Pathology, Chair of Clinical Pathology, Excellence Research Centre on Cardiovascular Disease, U.O.C. Immunohematology, Transfusion Medicine and Transplant Immunology (SIMT), Regional Reference Laboratory of Transplant Immunology (LIT), Azienda Universitaria Policlinico (AOU), 1st School of Medicine, Second University of Naples, Piazza Miraglia 2, 80138, Naples, Italy aut NATIONALLICENCE 700 1- Infante Teresa IRCCS SDN Foundation, Via Gianturco 113, 80134, Naples, Italy aut NATIONALLICENCE 700 1- Napoli Claudio Department of General Pathology, Chair of Clinical Pathology, Excellence Research Centre on Cardiovascular Disease, U.O.C. Immunohematology, Transfusion Medicine and Transplant Immunology (SIMT), Regional Reference Laboratory of Transplant Immunology (LIT), Azienda Universitaria Policlinico (AOU), 1st School of Medicine, Second University of Naples, Piazza Miraglia 2, 80138, Naples, Italy aut NATIONALLICENCE 773 0- Clinical and Translational Oncology Springer Milan 15/3(2013-03-01), 211-218 1699-048X 15:3<211 2013 15 12094 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer