title: [Comparison of the effect between procaine and 5-aza-dc on wnt inhibitory factor 1 promoter methylation of HepG2].
abstract: OBJECTIVE
  To observe the CpG island methylation status, mRNA and protein expression of the Wnt inhibitory factor-1 (Wif-1) gene with procaine or 5'-Aza-2'-deoxycycytidine (5-aza-dc) on HepG2. And to explore the comparison of the demethylation with 5-aza-dc or procaine.
  
  
  METHODS
  HepG2 cells were treated with 5-aza-dc or procaine. Wif-1 mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). Wif-1 protein expression was detected by Western blot. The promoter methylation status of Wif-1 gene on treatment or not were detected by methylation-specific PCR (MSP).
  
  
  RESULTS
  (1) RT-PCR and Western blot exhibited that both Wif-1 mRNA and Wif-1 protein expression were positive in groups treated with procaine and 5-aza-dc and highly positive in the L0 cells group, but weakly even negative in the HepG2 cells without any treatment, the difference were statistically significant (P < 0.05) (procaine experimental group was higher than that of 5-aza-dc experimental group, P < 0.05). (2) The positive rates of the promoter hypermethylated in procaine and 5-aza-de groups were (14.41 +/- 3.37)% and (29.29 +/- 1.84)%. Both of the two groups showed a part of the de-methylation status (P < 0.05).
  
  
  CONCLUSIONS
  Both of procaine and 5-aza-dc can reverse the abnormal methylation of Wif-1 gene. Procaine can be more effective than conventional used 5-aza-dc and could be a new demethylation drug.
authors.affiliation: Department of Gastroenterology, the Second Affiliate Hospital of Chongqing Medical University, Chongqing 400010, China.
authors.forename: Man
authors.initials: M
authors.lastname: Hu
authors.forename: Li-Li
authors.initials: LL
authors.lastname: Dai
authors.forename: Wei-Zheng
authors.initials: WZ
authors.lastname: Zeng
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism*; D001374:Azacitidine / Q000494:pharmacology*; D018899:CpG Islands; D019175:DNA Methylation; D056945:Hep G2 Cells; D006801:Humans; D011343:Procaine / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger; D012097:Repressor Proteins / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D012333:RNA, Messenger; D012097:Repressor Proteins; C485569:WIF1 protein, human; D011343:Procaine; D001374:Azacitidine