Silvia Fittipaldi1, Francesco Vasuri2, Sonia Bonora1, Alessio Degiovanni1, Giacomo Santandrea1, Alessandro Cucchetti3, Laura Gramantieri4, Luigi Bolondi4, Antonia D'Errico1. 1. "F. Addarii" Institute of Oncology and Transplant Pathology, Department of Specialty, Diagnostic and Experimental Medicine (DIMES), S. Orsola-Malpighi University Hospital, V.le Ercolani 4/2, 40138, Bologna, Italy. 2. "F. Addarii" Institute of Oncology and Transplant Pathology, Department of Specialty, Diagnostic and Experimental Medicine (DIMES), S. Orsola-Malpighi University Hospital, V.le Ercolani 4/2, 40138, Bologna, Italy. vasurifrancesco@libero.it. 3. Department of Medical and Surgical Sciences (DIMEC), S. Orsola-Malpighi University Hospital, Bologna, Italy. 4. Division of Internal Medicine, Department of Medical and Surgical Sciences, University of Bologna, S. Orsola-Malpighi University Hospital, Bologna, Italy.
Abstract
BACKGROUND: miRNA deregulation and vascular modifications constitute promising predictors in the study of hepatocellular carcinoma (HCC). In the literature, the relative miRNA abundance in HCC is usually determined using as control non-matched tumoral tissue, healthy liver, or cirrhotic liver. However, a common standard RNA control for the normalization toward the tissue gene expression was not settled yet. AIM: To assess the differences existing in the quantitative miRNA gene expression in HCC on tissue according to two different liver controls. METHODS: A wide array of miRNAs was analyzed on 22 HCCs arisen in cirrhotic and non-cirrhotic livers by means of microfluidic cards. Control samples included total RNA extracted from healthy and cirrhotic livers. Immunohistochemistry for CD34 and Nestin was performed to assess the pattern of intratumoral vascular modifications. RESULTS: Six miRNAs were deregulated in HCCs using either controls: miR-532, miR-34a, miR-93, miR-149#, miR-7f-2#, and miR-30a-5p. Notably, the miRNA expression changed significantly between HCCs arisen in cirrhotic and non-cirrhotic livers, according to the control used for normalization. Different miRNA profiles were found also in HCCs with different vascular patterns, according to the control used for normalization. CONCLUSIONS: Our data confirm that the choice of the methodology, and particularly the control used for normalization, represents the main concern in miRNA evaluation, particularly in a heterogeneous model such as liver pathology. Still we observed the deregulation of some common miRNAs as promising in HCC cancerogenesis and progression. A standardized control will be a crucial achievement to compare miRNA expression among different laboratories.
BACKGROUND: miRNA deregulation and vascular modifications constitute promising predictors in the study of hepatocellular carcinoma (HCC). In the literature, the relative miRNA abundance in HCC is usually determined using as control non-matched tumoral tissue, healthy liver, or cirrhotic liver. However, a common standard RNA control for the normalization toward the tissue gene expression was not settled yet. AIM: To assess the differences existing in the quantitative miRNA gene expression in HCC on tissue according to two different liver controls. METHODS: A wide array of miRNAs was analyzed on 22 HCCs arisen in cirrhotic and non-cirrhotic livers by means of microfluidic cards. Control samples included total RNA extracted from healthy and cirrhotic livers. Immunohistochemistry for CD34 and Nestin was performed to assess the pattern of intratumoral vascular modifications. RESULTS: Six miRNAs were deregulated in HCCs using either controls: miR-532, miR-34a, miR-93, miR-149#, miR-7f-2#, and miR-30a-5p. Notably, the miRNA expression changed significantly between HCCs arisen in cirrhotic and non-cirrhotic livers, according to the control used for normalization. Different miRNA profiles were found also in HCCs with different vascular patterns, according to the control used for normalization. CONCLUSIONS: Our data confirm that the choice of the methodology, and particularly the control used for normalization, represents the main concern in miRNA evaluation, particularly in a heterogeneous model such as liver pathology. Still we observed the deregulation of some common miRNAs as promising in HCC cancerogenesis and progression. A standardized control will be a crucial achievement to compare miRNA expression among different laboratories.
Authors: Sylvain Pradervand; Johann Weber; Jérôme Thomas; Manuel Bueno; Pratyaksha Wirapati; Karine Lefort; G Paolo Dotto; Keith Harshman Journal: RNA Date: 2009-01-28 Impact factor: 4.942
Authors: Laura Gramantieri; Manuela Ferracin; Francesca Fornari; Angelo Veronese; Silvia Sabbioni; Chang-Gong Liu; George A Calin; Catia Giovannini; Eros Ferrazzi; Gian Luca Grazi; Carlo M Croce; Luigi Bolondi; Massimo Negrini Journal: Cancer Res Date: 2007-07-01 Impact factor: 12.701
Authors: Jinmai Jiang; Yuriy Gusev; Ileana Aderca; Teresa A Mettler; David M Nagorney; Daniel J Brackett; Lewis R Roberts; Thomas D Schmittgen Journal: Clin Cancer Res Date: 2008-01-15 Impact factor: 12.531
Authors: Alexandra Drakaki; Maria Hatziapostolou; Christos Polytarchou; Christina Vorvis; George A Poultsides; John Souglakos; Vassilis Georgoulias; Dimitrios Iliopoulos Journal: BMC Cancer Date: 2015-07-24 Impact factor: 4.430