title: Knockdown of thrombomodulin enhances HCC cell migration through increase of ZEB1 and decrease of E-cadherin gene expression.
abstract: BACKGROUND
  Thrombomodulin (TM) is a key molecule mediating circulation homeostasis through its binding to thrombin. The TM-thrombin complex can activate protein C and thrombin-activatable fibrinolysis inhibitor to form a tight clot. In many cancer tissues, decrease of TM expression may correlate with cancer metastasis. However, the role of TM in hepatocellular carcinoma (HCC) progression is still unclear.
  
  
  METHODS
  We characterized TM expression in HCC cells (HepJ5 and skHep-1 cells) using real-time polymerase chain reaction (PCR) and Western blotting. We then manipulated TM expression using both TM-specific short hairpin RNA (shRNA) and overexpressing it in HCC cells. Transwell migration assay was performed to monitor the migratory ability of HCC cells under different levels of TM expression.
  
  
  RESULTS
  We found that TM was ectopically highly expressed in skHep-1 at both transcriptional and translational levels. After silencing TM expression in skHep-1 cells, we found that metastatic capability was dramatically increased. Conversely, overexpression of TM in HepJ5 cells decreased metastatic ability. We investigated the possible mechanism and found that decreased TM-mediated enhancement of cell migration was dependent on upregulation of ZEB1, a repressor of E-cadherin.
  
  
  CONCLUSIONS
  TM may be a modulator of cancer metastasis in HCC. Downregulation of TM expression may increase ZEB1 and decrease E-cadherin levels.
authors.affiliation: Department of Surgery, Taipei Medical University, Taipei City, Taiwan.
authors.forename: Ming-Te
authors.initials: MT
authors.lastname: Huang
authors.forename: Po-Li
authors.initials: PL
authors.lastname: Wei
authors.forename: Jun-Jen
authors.initials: JJ
authors.lastname: Liu
authors.forename: Der-Zen
authors.initials: DZ
authors.lastname: Liu
authors.forename: Huang
authors.initials: H
authors.lastname: Huey-Chun
authors.forename: Jane
authors.initials: J
authors.lastname: An
authors.forename: Cheng-Chia
authors.initials: CC
authors.lastname: Wu
authors.forename: Chih-Hsiung
authors.initials: CH
authors.lastname: Wu
authors.forename: Yuan-Soon
authors.initials: YS
authors.lastname: Ho
authors.forename: Yi-Yuan
authors.initials: YY
authors.lastname: Yang
authors.forename: Yu-Jia
authors.initials: YJ
authors.lastname: Chang
mesh_terms: D015153:Blotting, Western; D015820:Cadherins / Q000235:genetics / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D002465:Cell Movement; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018180:Thrombomodulin / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D000071799:Zinc Finger E-box-Binding Homeobox 1
chemical_list: D015820:Cadherins; D018398:Homeodomain Proteins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D018180:Thrombomodulin; D014157:Transcription Factors; C486307:ZEB1 protein, human; D000071799:Zinc Finger E-box-Binding Homeobox 1