title: The silencing of RECK gene is associated with promoter hypermethylation and poor survival in hepatocellular carcinoma.
abstract: BACKGROUND
  To evaluate the promoter methylation status of RECK gene and mRNA expression in patients with hepatocellular carcinoma (HCC).
  
  
  METHODS
  We analyzed RECK methylation by MSP, and RECK mRNA by real-time PCR in 74 HCC. The liver cell lines (7721, Chang and Hep-G2) were treated with 5-Aza-CdR and TSA.
  
  
  RESULTS
  RECK mRNA were lower in HCC tissues (Mean (-∆Ct) = -3.29) than that in Non-Hcc tissues (Mean (-∆Ct) = -2.42). Expression of RECK was elevated in only 24 (32.43%) of the 74 HCC patients but decreased (-∆∆Ct<0) in 50 (67.57%) of the patients. RECK promoter was hypermethylated in 55.4% (41/74) of HCCs, and in only 17.6% (13/74) of Non-Hcc samples. RECK mRNA were lower in HCC patients with hypermethylation (∆MI>=0.5) (Mean (-∆∆Ct) = -1.75) than those with demethylation (∆MI<0.5) (Mean (-∆∆Ct) = 0.05), and there is a decreased tendency for RECK mRNA in HCC patients with promoter hypermethylation (p = 0.002). There was a significantly correlation found between RECK mRNA and poor survival after surgery. After treated by 5-Aza-CdR and TSA, we found that RECK mRNA induced different changes in 7721, Chang and Hep-G2 cells. And RECK demethylation also induced by epigenetic inhibitors.
  
  
  CONCLUSIONS
  The results suggested that the hypermethylation may lead to promoter silencing of RECK mRNA and associated with poor survival in HCC.
authors.affiliation: Clinical Oncology Laboratory, Changzhou Tumor Hospital, Soochow University, Changzhou, China. zhangchangsong@suda.edu.cn
authors.forename: Changsong
authors.initials: C
authors.lastname: Zhang
authors.forename: Yang
authors.initials: Y
authors.lastname: Ling
authors.forename: Chenghui
authors.initials: C
authors.lastname: Zhang
authors.forename: Yun
authors.initials: Y
authors.lastname: Xu
authors.forename: Lu
authors.initials: L
authors.lastname: Gao
authors.forename: Rong
authors.initials: R
authors.lastname: Li
authors.forename: Jing
authors.initials: J
authors.lastname: Zhu
authors.forename: Lieying
authors.initials: L
authors.lastname: Fan
authors.forename: Lixin
authors.initials: L
authors.lastname: Wei
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000235:genetics*; D045744:Cell Line, Tumor; D019175:DNA Methylation / Q000235:genetics*; D058851:GPI-Linked Proteins / Q000235:genetics*; D020868:Gene Silencing; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D058851:GPI-Linked Proteins; C115460:RECK protein, human