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Journal of Hepatology 51 (2009) 114–126 www.elsevier.com/locate/jhep

Identification of proteomic changes during human liver development by 2D-DIGE and mass spectrometryq
´ Jean Paul Brizard1, Jeanne Ramos2, Agnes Robert3, Daniel Lafitte4, Nicole Bigi5, 5 6 Pierre Sarda , Dalila Laoudj-Chenivesse , Francis Navarro7, Pierre Blanc7, Eric Assenat7,8, ´ Patrick Maurel3, Jean-Marc Pascussi3,Marie-Jose Vilarem3,*
´ ´ Institut de Recherche pour le Developpement (IRD), UMR 5096 (CNRS-IRD-Universite Perpignan), Montpellier, France 2 Service d’Anatomo-pathologie, CHU Gui de Chauliac, Montpellier, France 3 Inserm, UMR-S632, University Montpellier 1, 34293 Montpellier, France 4 ´ Faculte de Pharmacie, Marseille, France 5 ˆ ´ ´ ´ ´ ´ ´ Centre de Reference Anomalies du Developpement et SyndromesMalformatifs, Service de Genetique Medicale, Hopital Arnaud de Villeneuve, ´ Centre Hospitalier Regional et Universitaire, Montpellier, France 6 ˆ INSERM ERI 25, Hopital Arnaud de Villeneuve, Montpellier, France 7 ´ Service Medico-Chirugical des Maladies de l’Appareil Digestif et de Transplantation Hepatique, CHU Saint Eloi, Montpellier, France 8 Service d’Oncologie Digestive, CRLC Paul Lamarque, Vald’Aurelle, Montpellier, France
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Background/Aims: The aim of this study was to identify human liver proteins that are associated with different stages of liver development. Methods: We collected liver samples from 14 fetuses between 14 and 41 weeks of development, one child and four adults. Proteins which exhibited consistent and significant variations during development by two-dimensionaldifferential in gel electrophoresis (2D-DIGE) were subjected to peptide mass fingerprint analysis by MALDI-TOF mass spectrometry. Real-time PCR analysis confirmed, at the transcriptional level, the data obtained by the proteomic approach. Results: Among a total of 80 protein spots showing differential expression, we identified 42 different proteins or polypeptide chains, of which 26 were upregulated and 16downregulated in developing in comparison to adult liver. These proteins could be classified in specific groups according to their function. By comparing their temporal expression profiles, we identified protein groups that were associated with different developmental stages of human fetal liver and suggest that the changes in protein expression observed during the 20- to 36-week time window play apivotal role in liver development. Conclusions: The identification of these proteins may represent good markers of human liver and stem cells differentiation. Ó 2009 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved. Keywords: Liver development; Hepatocellular carcinoma; Two-dimensional differential in gel electrophoresis

1. Introduction
Received 23October 2008; received in revised form 6 February 2009; accepted 18 February 2009; available online 17 April 2009 Associate Editor: C. Trautwein q The authors who have taken part in this study declared that they do not have anything to disclose regarding funding from industry or conflict of interest with respect to this manuscript. * Corresponding author. Tel.: +33 4 67613369; fax: +33 4 67523681. E-mailaddress: marie-jose.vilarem@inserm.fr (M.-J. Vilarem). Abbreviations: HCC, hepatocellular carcinoma; 2D-DIGE, twodimensional differential in gel electrophoresis; MS, MALDI-TOF mass.

The liver is the largest organ of the body and plays a major role in energy metabolism, protein synthesis, homeostasis, blood proteins secretion, detoxification in the adult, and in hematopoiesis in the embryo. It iscomposed of a heterogeneous cell population that includes hepatocytes, non-parenchymal stellate and ovals cells, macrophages, vascular/sinusoidal endothelial cells and

0168-8278/$36.00 Ó 2009 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved. doi:10.1016/j.jhep.2009.02.029

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