naa a22 4500 510773648 CHVBK 20180411083223.0 cr unu---uuuuu 180411e20130601xx s 000 0 eng 10.1007/s12022-013-9246-y doi (NATIONALLICENCE)springer-10.1007/s12022-013-9246-y Characterization of Neuroendocrine Tumors of the Pancreas by Real-Time Quantitative Polymerase Chain Reaction. A Methodological Approach [Elektronische Daten] [Laura Annaratone, Marco Volante, Sofia Asioli, Nelson Rangel, Gianni Bussolati] The aim of this study was to assess the suitability of using real-time quantitative PCR (RT-qPCR) to characterize neuroendocrine (NE) tumors of the pancreas. For a series of tumors, we evaluated several genes of interest, and the data were matched with the "classical” immunohistochemical (IHC) features. In 21 cases, we extracted RNA from formalin-fixed paraffin-embedded (FFPE) blocks, and in nine cases, we also extracted RNA from fresh-frozen tissue. The RT-qPCR procedure was performed using two sets of customized arrays. The test using the first set, covering 96 genes of interest, was focused on assessing the feasibility of the procedure, and the results were used to select 18 genes indicative of NE differentiation, clinical behavior, and therapeutic responsiveness for use in the second set of arrays. Threshold cycle (Ct) values were used to calculate the fold-changes in gene expression using the 2-∆∆Ct method. Statistical procedures were used to analyze the results, which were matched with the IHC and follow-up data. Material from fresh-frozen samples performed better in terms of the level of amplification, but acceptable and concordant results were also obtained from FFPE samples. In addition, high concordance was observed between the mRNA and protein expression levels of somatostatin receptor type 2A (R = 0.52, p = 0.016). Genes associated with NE differentiation, as well as the gastrin-releasing peptide receptor and O-6-methylguanine-DNA methyltransferase genes, were underexpressed, whereas angiogenesis-associated markers (CDH13 and SLIT2) were overexpressed in tissues with malignant behavior. The RT-qPCR procedure is practical and feasible in economic terms for the characterization of NE tumors of the pancreas and can complement morphological and IHC-based evaluations. Thus, the results of the RT-qPCR procedure might offer an objective basis for therapeutic choices. Springer Science+Business Media New York, 2013 Pancreas nationallicence Neuroendocrine neoplasm nationallicence Gene expression nationallicence M : Male nationallicence F : Female nationallicence WHO : World Health Organization classification nationallicence NET : Neuroendocrine tumor nationallicence NEC : Neuroendocrine carcinoma nationallicence G : Grade nationallicence IHC : Immunohistochemistry nationallicence T : Tumor nationallicence N : Lymph node nationallicence M : Metastasis nationallicence SSTR2A : Somatostatin receptor subtype 2A nationallicence p-mTOR : Phosphorylated mammalian target of rapamycin nationallicence NED : Not evidence of disease nationallicence AWD : Alive with disease nationallicence DOD : Dead of disease nationallicence *Hormone IHC : Hormonal status detected by the immunohistochemical analysis of paraffin-embedded tissue nationallicence Annaratone Laura Department of Medical Sciences, University of Turin, Via Santena 7, 10126, Turin, Italy aut Volante Marco Department of Oncology, University of Turin, Turin, Italy aut Asioli Sofia Department of Medical Sciences, University of Turin, Via Santena 7, 10126, Turin, Italy aut Rangel Nelson Department of Medical Sciences, University of Turin, Via Santena 7, 10126, Turin, Italy aut Bussolati Gianni Department of Medical Sciences, University of Turin, Via Santena 7, 10126, Turin, Italy aut Endocrine Pathology Springer US; http://www.springer-ny.com 24/2(2013-06-01), 83-91 1046-3976 24:2<83 2013 24 12022 https://doi.org/10.1007/s12022-013-9246-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s12022-013-9246-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Annaratone Laura Department of Medical Sciences, University of Turin, Via Santena 7, 10126, Turin, Italy aut NATIONALLICENCE 700 1- Volante Marco Department of Oncology, University of Turin, Turin, Italy aut NATIONALLICENCE 700 1- Asioli Sofia Department of Medical Sciences, University of Turin, Via Santena 7, 10126, Turin, Italy aut NATIONALLICENCE 700 1- Rangel Nelson Department of Medical Sciences, University of Turin, Via Santena 7, 10126, Turin, Italy aut NATIONALLICENCE 700 1- Bussolati Gianni Department of Medical Sciences, University of Turin, Via Santena 7, 10126, Turin, Italy aut NATIONALLICENCE 773 0- Endocrine Pathology Springer US; http://www.springer-ny.com 24/2(2013-06-01), 83-91 1046-3976 24:2<83 2013 24 12022 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer