title: A supercritical-CO2 extract of Ganoderma lucidum spores inhibits cholangiocarcinoma cell migration by reversing the epithelial-mesenchymal transition.
abstract: BACKGROUND
  Ganoderma lucidum (G. lucidum) is an oriental medical mushroom that has been widely used in Asian countries for centuries to prevent and treat different diseases, including cancer.
  
  
  OBJECTIVE
  The objective of this study was to investigate the effect of A supercritical-CO2 extract of G. lucidum spores on the transforming growth factor beta 1 (TGF-β1)-induced epithelial-mesenchymal transition (EMT) of cholangiocarcinoma cells.
  
  
  METHODS
  This was an in vitro study with human cholangiocarcinoma TFK-1 cells treated with varying concentrations of G. lucidum.
  
  
  METHODS
  A supercritical-CO2 extract of G. lucidum spores (GLE) was obtained from completely sporoderm-broken germinating G. lucidum spores by supercritical fluid carbon dioxide (SCF-CO2) extraction. GLE pre-incubated with human cholangiocarcinoma TFK-1 cells prior to TGF-β1 treatment (2ng/ml) for 48h. Changes in EMT markers were analyzed by western blotting and immunofluorescence. The formation of F-actin stress fibers was assessed via immunostaining with phalloidin and examined using confocal microscopy. Additionally, the effect of the GLE on TGF-β1-induced migration was investigated by a Boyden chamber assay.
  
  
  RESULTS
  TGF-β1-induced reduction in E-cadherin expression was associated with a loss of epithelial morphology and cell-cell contact. Concomitant increases in N-cadherin and Fibronectin were evident in predominantly elongated fibroblast-like cells. The GLE suppressed the TGF-β1-induced morphological changes and the changes in cadherin expression, and also inhibited the formation of F-actin stress fibers, which are a hallmark of EMT. The GLE also inhibited TGF-β1-induced migration of TFK-1 cells.
  
  
  CONCLUSIONS
  Our findings provide new evidence that GLE suppress cholangiocarcinoma migration in vitro through inhibition of TGF-β1-induced EMT. The GLE may be clinically applied in the prevention and/or treatment of cancer metastasis.
authors.affiliation: State Key Laboratory of Biocontrol, Sun Yat-sen (Zhongshan) University, Guangzhou, Guangdong, PR China.
authors.forename: Lian
authors.initials: L
authors.lastname: Li
authors.affiliation: Basic Medical College Jiangxi University of traditional Chinese Medicine, Nanchang, Jiangxi, PR China.
authors.forename: Hui-Jun
authors.initials: HJ
authors.lastname: Guo
authors.affiliation: State Key Laboratory of Biocontrol, Sun Yat-sen (Zhongshan) University, Guangzhou, Guangdong, PR China.
authors.forename: Ling-Yan
authors.initials: LY
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Biocontrol, Sun Yat-sen (Zhongshan) University, Guangzhou, Guangdong, PR China.
authors.forename: Limin
authors.initials: L
authors.lastname: Zheng
authors.affiliation: Academy of Food and Health Engineering, Sun Yat-Sen University, Guangzhou, Guangdong, PR China. Electronic address: lsslx@mail.sysu.edu.cn.
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
mesh_terms: D000199:Actins / Q000378:metabolism; D015703:Antigens, CD / Q000378:metabolism; D001688:Biological Products / Q000494:pharmacology*; D015820:Cadherins / Q000378:metabolism; D045744:Cell Line, Tumor / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects*; D018281:Cholangiocarcinoma / Q000473:pathology*; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects*; D005353:Fibronectins / Q000378:metabolism; D006801:Humans; D020940:Reishi / Q000737:chemistry*; D013172:Spores, Fungal / Q000737:chemistry; D053773:Transforming Growth Factor beta1; D014315:Triterpenes / Q000494:pharmacology
chemical_list: D000199:Actins; D015703:Antigens, CD; D001688:Biological Products; C050361:CDH2 protein, human; D015820:Cadherins; D005353:Fibronectins; D053773:Transforming Growth Factor beta1; D014315:Triterpenes
keywords: Cholangiocarcinoma TFK-1; Epithelial–mesenchymal transition; Ganoderma lucidum; Migration; Supercritical-CO2 extract