title: [Methylation of matrix metalloproteinases-2 promoter in endometrial cancer invasion].
abstract: OBJECTIVE
  To investigate the regulation of DNA methylation on the expression of MMP-2 gene, and the effect of 5-aza-2'-deoxycytidine (5-aza-CdR) on invasiveness of endometrial cancer cells.
  
  
  METHODS
  Human endometrial cancer cell lines Ishikawa and HEC-1A were treated with 5-aza-CdR. After genomic DNA was extracted from the cells and treated with bisulfate sodium, clone sequencing was used to detect methylation of the matrix  metalloproteinases-2 (MMP-2) promoter region. Ishikawa and HEC-1A cells were treated with different concentrations of 5-aza-CdR, and the effect  on the invasion and metastasis to the endometrial cancer (EC) cells were detected by Transwell invasion chamber. The MMP-2 mRNA expression was  detected by real-time quantitative PCR.
  
  
  RESULTS
  Although the MMP-2 promoter showed global hypomethylation status, the methylation of CG  dinucleotide sites located on the transcription factor binding sites had high positive frequencies, which were 80% and 70% in HEC-1A cells and 60% and  40% in Ishikawa cells, respectively. The methylation frequencies reduced to 20% and 10% in HEC-1A cells by demethylating agent 5-aza-CdR and the  CG sites in Ishikawa cells were totally demethylated. Moreover, after 72 h of treatment with different concentrations of 5-aza-CdR, the expression of the  MMP-2 mRNA and transmembrane number of both Ishikawa and HEC-1A cells were significantly upregulated compared with the control group (P<0.05).
  
  
  CONCLUSIONS
  Methylation of CG dinucleotide sites of the MMP-2 promotor region may be associated with the regulation of gene transcription. The 5-aza-CdR can promote the invasion and metastasis of endometrial cancer cells in vitro by upregulating the MMP-2 mRNA expression.
authors.affiliation: Department of Gynecology and Obstetrics, Peking University First Hospital, Beijing, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Hu
authors.forename: Li
authors.initials: L
authors.lastname: Yu
authors.forename: Qin-ping
authors.initials: QP
authors.lastname: Liao
mesh_terms: D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D019175:DNA Methylation / Q000235:genetics*; D000077209:Decitabine; D016889:Endometrial Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000235:genetics* / Q000378:metabolism; D009361:Neoplasm Invasiveness / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D012333:RNA, Messenger; D000077209:Decitabine; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D001374:Azacitidine