title: Changes in protein expression in two cholangiocarcinoma cell lines undergoing formation of multicellular tumor spheroids in vitro.
abstract: Epithelial-to-Mesenchymal Transition (EMT) is relevant in malignant growth and frequently correlates with worsening disease progression due to its implications in metastases and resistance to therapeutic interventions. Although EMT is known to occur in several types of solid tumors, the information concerning tumors arising from the epithelia of the bile tract is still limited. In order to approach the problem of EMT in cholangiocarcinoma, we decided to investigate the changes in protein expression occurring in two cell lines under conditions leading to growth as adherent monolayers or to formation of multicellular tumor spheroids (MCTS), which are considered culture models that better mimic the growth characteristics of in-vivo solid tumors. In our system, changes in phenotypes occur with only a decrease in transmembrane E-cadherin and vimentin expression, minor changes in the transglutaminase protein/activity but with significant differences in the proteome profiles, with declining and increasing expression in 6 and in 16 proteins identified by mass spectrometry. The arising protein patterns were analyzed based on canonical pathways and network analysis. These results suggest that significant metabolic rearrangements occur during the conversion of cholangiocarcinomas cells to the MCTS phenotype, which most likely affect the carbohydrate metabolism, protein folding, cytoskeletal activity, and tissue sensitivity to oxygen.
authors.affiliation: Department of Biomedical and Surgical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Mischiati
authors.affiliation: Institute for Maternal and Child Health, IRCCS Burlo Garofalo, University of Trieste, Trieste, Italy.
authors.forename: Blendi
authors.initials: B
authors.lastname: Ura
authors.affiliation: Center for Prevention and Diagnosis of Coeliac Disease/Gastroenterology 2, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, Milano, Italy; Department of Biomedical, Surgical and Odontoiatric Sciences, University of Milano, Milano, Italy.
authors.forename: Leda
authors.initials: L
authors.lastname: Roncoroni
authors.affiliation: Center for Prevention and Diagnosis of Coeliac Disease/Gastroenterology 2, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, Milano, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Elli
authors.affiliation: Department of Biomedical and Surgical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Cervellati
authors.affiliation: Department of Biomedical and Surgical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Squerzanti
authors.affiliation: Center for Prevention and Diagnosis of Coeliac Disease/Gastroenterology 2, Fondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico, Milano, Italy.
authors.forename: Dario
authors.initials: D
authors.lastname: Conte
authors.affiliation: Department of Biomedical, Surgical and Odontoiatric Sciences, University of Milano, Milano, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Doneda
authors.affiliation: CRIBI Centre of Innovative Biotechnologies, University of Padua, Padua, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Polverino de Laureto
authors.affiliation: CRIBI Centre of Innovative Biotechnologies, University of Padua, Padua, Italy.
authors.forename: Giorgia
authors.initials: G
authors.lastname: de Franceschi
authors.affiliation: Department of Biomedical and Surgical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Calza
authors.affiliation: Moulder Center for Drug Discovery Research, Temple University, School of Pharmacy, Philadelphia, Pennsylvania, United States of America.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Barrero
authors.affiliation: Moulder Center for Drug Discovery Research, Temple University, School of Pharmacy, Philadelphia, Pennsylvania, United States of America.
authors.forename: Salim
authors.initials: S
authors.lastname: Merali
authors.affiliation: Department of Clinical and Molecular Sciences, Faculty of Medicine, Le Marche Polytechnic University, Ancona, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Ferrari
authors.affiliation: Department of Biomedical and Surgical Sciences, University of Ferrara, Ferrara, Italy.
authors.forename: Carlo M
authors.initials: CM
authors.lastname: Bergamini
authors.affiliation: Istituto Pasteur, Fondazione Cenci Bolognetti and Department of Biochemical Sciences "A. Rossi Fanelli", La Sapienza University of Rome and CNR, Biology and Molecular Pathology Institutes, Rome, Italy.
authors.forename: Enzo
authors.initials: E
authors.lastname: Agostinelli
mesh_terms: D001650:Bile Duct Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D018281:Cholangiocarcinoma / Q000378:metabolism* / Q000473:pathology; D058750:Epithelial-Mesenchymal Transition; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D018874:Spheroids, Cellular / Q000378:metabolism*
chemical_list: D020543:Proteome