title: Epigenetic-mediated upregulation of progesterone receptor B gene in endometrial cancer cell lines.
abstract: OBJECTIVE
  To determine if epigenetic interference can restore progesterone receptor-B (PR-B) expression in PR-B negative endometrial adenocarcinoma cell lines, and to characterize the kinetics of PR-B induction mediated by DNA methyltransferase and histone deacetylase inhibitors.
  
  
  METHODS
  The PR-B negative endometrioid cancer cell lines KLE and HEC-1B were used as study models. PR-B mRNA and protein expression levels were measured using real-time PCR and Western blot analysis, respectively. DNA methylation levels of the PR-B promoter were determined by methylation-specific PCR. Dose-response correlations and the duration of response to aza-deoxycytidine (ADC) and trichostatin A (TSA) were characterized. Cell responses to prolonged and repeated drug treatment were also examined.
  
  
  RESULTS
  Relatively low concentrations of ADC and TSA over a 24-h period induced PR-B expression. Furthermore, ADC and TSA acted synergistically to reactivate PR-B expression. Depending on the cell line used, PR-B mRNA was induced 10-110 fold. This elevated PR-B expression continued for 48 h after drug withdrawal. Sustained upregulation of PR-B mRNA and protein was observed during prolonged and repeated drug treatment.
  
  
  CONCLUSIONS
  The epigenetically silenced PR-B gene remains sensitive to changes in DNA demethylation and histone acetylation in uterine adenocarcinoma cell lines. Treatment with ADC and/or TSA results in a robust and sustainable PR-B upregulation. These small molecule epigenetic modifying agents may be used to sensitize poorly differentiated, PR-B negative endometrial cancers to progestational therapy.
authors: Department of Obstetrics and Gynecology, Mayo Clinic and Foundation, 200 First Street SW, Rochester, MN 55905, USA.
authors: Yuning
authors: Y
authors: Xiong
authors: Sean C
authors: SC
authors: Dowdy
authors: Jesus
authors: J
authors: Gonzalez Bosquet
authors: Ying
authors: Y
authors: Zhao
authors: Norman L
authors: NL
authors: Eberhardt
authors: Karl C
authors: KC
authors: Podratz
authors: Shi-Wen
authors: SW
authors: Jiang
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics* / Q000378:metabolism; D001374:Azacitidine / Q000031:analogs & derivatives / Q000494:pharmacology; D045744:Cell Line, Tumor; D019175:DNA Methylation; D000077209:Decitabine; D016889:Endometrial Neoplasms / Q000235:genetics* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D044127:Epigenesis, Genetic; D005260:Female; D056572:Histone Deacetylase Inhibitors; D006655:Histone Deacetylases / Q000378:metabolism; D006801:Humans; D006877:Hydroxamic Acids / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011980:Receptors, Progesterone / Q000096:biosynthesis / Q000235:genetics*; D015854:Up-Regulation / Q000187:drug effects
chemical_list: D004791:Enzyme Inhibitors; D056572:Histone Deacetylase Inhibitors; D006877:Hydroxamic Acids; D012333:RNA, Messenger; D011980:Receptors, Progesterone; C097314:progesterone receptor B; C012589:trichostatin A; D000077209:Decitabine; D006655:Histone Deacetylases; D001374:Azacitidine