title: Expression analysis of genes involved in epigenetic regulation and apoptosis in human malignant haematopoietic cell lines treated with 5-azacytidine.
abstract: OBJECTIVE
  The aim of this study was to investigate the modulation of the expression status of 10 different genes involved in epigenetic regulation and apoptosis by the DNA methyltransferase (DNMT) inhibitor 5-azacytidine (5-Aza), as markers of response to treatment, in two different human malignant haematopoietic cell lines.
  
  
  METHODS
  In our analysis we used the SybrGreen technology and gene-specific primers for the qRT-PCR analysis of 10 genes, in cDNA of PC-MDS and K562 cell lines, treated by 1 micromole of 5-Aza for 24h.
  
  
  RESULTS
  DNMT1 and DNMT3A showed statistically significant decrease of expression in 5-Aza-treated PC-MDS cells, whereas DNMT3B showed significantly decreased expression in 5-Aza-treated K562 cells. The members of the Bcl- 2 family of apoptosis-regulating genes Bcl-2 and Bax showed statistically significant differences in expression, in comparison with non-treated PC-MDS cells. Our most interesting result was the significant upregulation (re-expression) of p15, in 5-Aza-treated PC-MDS cells.
  
  
  CONCLUSIONS
  The re-expression of p15 in PC-MDS cell line evaluated by qRT-PCR makes this novel cell line a suitable model for the studies of pharmacologic demethylation as a plausible mechanism resulting in hematologic response in myelodysplastic syndrome (MDS).
authors.affiliation: Department of Biochemistry, Medical Faculty, Clinical Center of Vojvodina, University of Novi Sad, Novi Sad, Serbia. stankovkarmen@yahoo.com
authors.forename: K
authors.initials: K
authors.lastname: Stankov
authors.forename: G
authors.initials: G
authors.lastname: Bogdanovic
authors.forename: V
authors.initials: V
authors.lastname: Kojic
authors.forename: S
authors.initials: S
authors.lastname: Stankov
authors.forename: N
authors.initials: N
authors.lastname: Jojic
authors.forename: L
authors.initials: L
authors.lastname: Popovic
authors.forename: S
authors.initials: S
authors.lastname: Popovic
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D017209:Apoptosis; D001374:Azacitidine / Q000494:pharmacology*; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics; D000090205:DNA Methyltransferase 3A; D044127:Epigenesis, Genetic; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D020014:K562 Cells; D009190:Myelodysplastic Syndromes / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D018384:Oxidative Stress; D012333:RNA, Messenger / Q000032:analysis
chemical_list: D000964:Antimetabolites, Antineoplastic; C000717967:DNMT3A protein, human; D012333:RNA, Messenger; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; C000619380:DNMT1 protein, human; D001374:Azacitidine