title: Novel effect of antihelminthic Niclosamide on S100A4-mediated metastatic progression in colon cancer.
abstract: BACKGROUND
  Metastasis formation in colon cancer severely reduces the survival rate in patients. S100A4, a calcium-binding protein, is implicated in promoting metastasis formation in colon cancer.
  
  
  METHODS
  To identify a transcription inhibitor of S100A4, high-throughput screening of 1280 pharmacologically active compounds was performed using a human colon cancer cell line expressing a S100A4 promoter-driven luciferase (LUC) reporter gene construct (HCT116-S1004p-LUC). Niclosamide, an antihelminthic agent, was identified as a potential candidate. Colon cancer cell lines (HCT116, SW620, LS174T, SW480, and DLD-1) were treated with 1 μM niclosamide to analyze the effect on S100A4 mRNA and protein expression by quantitative reverse transcription-polymerase chain reaction and immunoblot assays, and effects on cell migration, invasion, proliferation, and colony formation were also assessed in vitro. The effect of niclosamide on liver metastasis was assessed in a xenograft mouse model of human colon cancer (n = 8 mice) by in vivo imaging. The long-term effect of niclosamide on metastasis formation after discontinued treatment was quantified by scoring, and overall survival (n = 12 mice) was analyzed by Kaplan-Meier method after discontinuation of treatment. All statistical tests were two-sided.
  
  
  RESULTS
  Reduced S100A4 mRNA and protein expression, and inhibited cell migration, invasion, proliferation, and colony formation were observed in niclosamide-treated colon cancer cells in vitro. In vivo imaging of niclosamide-treated mice showed reduced liver metastasis compared with solvent-treated control mice (n = 4 mice per group). Compared with the control group, discontinuation of treatment for 26 days showed reduced liver metastasis formation in mice (n = 6 mice per group) (control vs discontinuous treatment, mean metastasis score = 100% vs 34.9%, mean difference = 65.1%; 95% confidence interval [CI] = 18.4% to 111.9%, P < .01) and increased overall survival (n = 6 mice per group; control vs discontinuous treatment, median survival = 24 vs 46.5 days, ratio = 0.52, 95% CI = 0.19 to 0.84, P = .001).
  
  
  CONCLUSIONS
  Niclosamide inhibits S100A4-induced metastasis formation in a mouse model of colon cancer and has therapeutic potential.
authors.affiliation: Max-Delbrück-Center for Molecular Medicine, Berlin, Germany.
authors.forename: Ulrike
authors.initials: U
authors.lastname: Sack
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Walther
authors.forename: Dominic
authors.initials: D
authors.lastname: Scudiero
authors.forename: Mike
authors.initials: M
authors.lastname: Selby
authors.forename: Dennis
authors.initials: D
authors.lastname: Kobelt
authors.forename: Margit
authors.initials: M
authors.lastname: Lemm
authors.forename: Iduna
authors.initials: I
authors.lastname: Fichtner
authors.forename: Peter M
authors.initials: PM
authors.lastname: Schlag
authors.forename: Robert H
authors.initials: RH
authors.lastname: Shoemaker
authors.forename: Ulrike
authors.initials: U
authors.lastname: Stein
mesh_terms: D000818:Animals; D000871:Anthelmintics / Q000494:pharmacology; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D047369:Chromatin Immunoprecipitation; D003110:Colonic Neoplasms / Q000188:drug therapy* / Q000378:metabolism* / Q000473:pathology; D018450:Disease Progression; D024202:Electrophoretic Mobility Shift Assay; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D015151:Immunoblotting; D007263:Infusions, Parenteral; D053208:Kaplan-Meier Estimate; D008113:Liver Neoplasms / Q000517:prevention & control* / Q000556:secondary; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D009361:Neoplasm Invasiveness; D009534:Niclosamide / Q000008:administration & dosage / Q000737:chemistry / Q000494:pharmacology*; D011379:Prognosis; D012333:RNA, Messenger / Q000378:metabolism; D012334:RNA, Neoplasm / Q000378:metabolism; D011897:Random Allocation; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013997:Time Factors; D014183:Transplantation, Heterologous; D014410:Tumor Stem Cell Assay; D051153:Wnt Proteins / Q000187:drug effects / Q000378:metabolism; D051176:beta Catenin / Q000187:drug effects / Q000378:metabolism
chemical_list: D000871:Anthelmintics; D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; C495270:CTNNB1 protein, human; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D000071999:S100 Calcium-Binding Protein A4; D009418:S100 Proteins; D051153:Wnt Proteins; D051176:beta Catenin; C077591:S100A4 protein, human; D009534:Niclosamide