title: Low-Dose Paclitaxel Reduces S100A4 Nuclear Import to Inhibit Invasion and Hematogenous Metastasis of Cholangiocarcinoma.
abstract: Nuclear expression of the calcium-binding protein S100A4 is a biomarker of increased invasiveness in cholangiocarcinoma, a primary liver cancer with scarce treatment opportunities and dismal prognosis. In this study, we provide evidence that targeting S100A4 nuclear import by low-dose paclitaxel, a microtubule-stabilizing agent, inhibits cholangiocarcinoma invasiveness and metastatic spread. Administration of low-dose paclitaxel to established (EGI-1) and primary (CCA-TV3) cholangiocarcinoma cell lines expressing nuclear S100A4 triggered a marked reduction in nuclear expression of S100A4 without modifying its cytoplasmic levels, an effect associated with a significant decrease in cell migration and invasiveness. While low-dose paclitaxel did not affect cellular proliferation, apoptosis, or cytoskeletal integrity, it significantly reduced SUMOylation of S100A4, a critical posttranslational modification that directs its trafficking to the nucleus. This effect of low-dose paclitaxel was reproduced by ginkolic acid, a specific SUMOylation inhibitor. Downregulation of nuclear S100A4 by low-dose paclitaxel was associated with a strong reduction in RhoA and Cdc42 GTPase activity, MT1-MMP expression, and MMP-9 secretion. In an SCID mouse xenograft model, low-dose metronomic paclitaxel treatment decreased lung dissemination of EGI-1 cells without significantly affecting their local tumor growth. In the tumor mass, nuclear S100A4 expression by cholangiocarcinoma cells was significantly reduced, whereas rates of proliferation and apoptosis were unchanged. Overall, our findings highlight nuclear S100A4 as a candidate therapeutic target in cholangiocarcinoma and establish a mechanistic rationale for the use of low-dose paclitaxel in blocking metastatic progression of cholangiocarcinoma. Cancer Res; 76(16); 4775-84. ©2016 AACR.
authors.affiliation: School of Medicine and Surgery, University of Milan-Bicocca, Milano, Italy. International Center for Digestive Health (ICDH), University of Milan-Bicocca, Milano, Italy.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Cadamuro
authors.affiliation: School of Medicine and Surgery, University of Milan-Bicocca, Milano, Italy.
authors.forename: Gaia
authors.initials: G
authors.lastname: Spagnuolo
authors.affiliation: Metabolism, Disease and Clinical Nutrition Unit, Treviso Regional Hospital, Treviso, Italy.
authors.forename: Luisa
authors.initials: L
authors.lastname: Sambado
authors.affiliation: Veneto Institute of Oncology IOV-IRCCS, Padua, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Indraccolo
authors.affiliation: Veneto Institute of Oncology IOV-IRCCS, Padua, Italy.
authors.forename: Giorgia
authors.initials: G
authors.lastname: Nardo
authors.affiliation: Veneto Institute of Oncology IOV-IRCCS, Padua, Italy. Department of Surgery, Oncology and Gastroenterology, University of Padua School of Medicine, Padua, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Rosato
authors.affiliation: School of Medicine and Surgery, University of Milan-Bicocca, Milano, Italy.
authors.forename: Simone
authors.initials: S
authors.lastname: Brivio
authors.affiliation: School of Medicine and Surgery, University of Milan-Bicocca, Milano, Italy.
authors.forename: Chiara
authors.initials: C
authors.lastname: Caslini
authors.affiliation: 4 Surgery Division, Treviso Regional Hospital, Treviso, Italy.
authors.forename: Tommaso
authors.initials: T
authors.lastname: Stecca
authors.affiliation: 4 Surgery Division, Treviso Regional Hospital, Treviso, Italy.
authors.forename: Marco
authors.initials: M
authors.lastname: Massani
authors.affiliation: Department of Surgery, Oncology and Gastroenterology, University of Padua School of Medicine, Padua, Italy. 4 Surgery Division, Treviso Regional Hospital, Treviso, Italy.
authors.forename: Nicolò
authors.initials: N
authors.lastname: Bassi
authors.affiliation: Biostatistics Unit, Clinica San Gaudenzio, Novara, Italy.
authors.forename: Eugenio
authors.initials: E
authors.lastname: Novelli
authors.affiliation: International Center for Digestive Health (ICDH), University of Milan-Bicocca, Milano, Italy. Section of Digestive Diseases, Yale University School of Medicine, New Haven, Connecticut.
authors.forename: Carlo
authors.initials: C
authors.lastname: Spirli
authors.affiliation: International Center for Digestive Health (ICDH), University of Milan-Bicocca, Milano, Italy. Section of Digestive Diseases, Yale University School of Medicine, New Haven, Connecticut. Department of Molecular Medicine, University of Padua School of Medicine, Padua, Italy. luca.fabris@unipd.it.
authors.forename: Luca
authors.initials: L
authors.lastname: Fabris
authors.affiliation: School of Medicine and Surgery, University of Milan-Bicocca, Milano, Italy. International Center for Digestive Health (ICDH), University of Milan-Bicocca, Milano, Italy. Section of Digestive Diseases, Yale University School of Medicine, New Haven, Connecticut.
authors.forename: Mario
authors.initials: M
authors.lastname: Strazzabosco
mesh_terms: D021581:Active Transport, Cell Nucleus / Q000187:drug effects*; D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D001650:Bile Duct Neoplasms / Q000473:pathology*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D018281:Cholangiocarcinoma / Q000473:pathology*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D009361:Neoplasm Invasiveness / Q000473:pathology; D009362:Neoplasm Metastasis; D017239:Paclitaxel / Q000494:pharmacology*; D000071999:S100 Calcium-Binding Protein A4 / Q000378:metabolism*; D058207:Sumoylation / Q000187:drug effects; D023041:Xenograft Model Antitumor Assays
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D000071999:S100 Calcium-Binding Protein A4; C077591:S100A4 protein, human; D017239:Paclitaxel