title: Estrogen-receptor-dependent regulation of telomerase activity in human endometrial cancer cell lines.
abstract: OBJECTIVE
  Given that prolonged exposure to unopposed estrogen is associated with endometrial cancer development and that the promoter region of the catalytic subunit of the telomerase enzyme, hTERT, contains putative estrogen response elements (EREs), we postulated that estrogen-receptor (ER)-mediated induction of telomerase activity may play an important role in endometrial carcinogenesis.
  
  
  METHODS
  ER-positive and ER-negative endometrial cancer cell lines were used. ER alpha expression was reconstituted in ER-negative cell lines by transient transfection. Telomerase activity was assayed using a PCR-based telomeric repeat amplification protocol (TRAP) after exposure to estradiol (E2). hTERT mRNA expression was assessed by real-time RT-PCR. Gel shift assays using oligonucleotide probes encoding each ERE and transient expression assays using luciferase reporter plasmids containing varying lengths of the 5' promoter region of the hTERT gene were performed.
  
  
  RESULTS
  E2 induced both hTERT gene transcription and telomerase activity in the ER-positive cell lines, but not in the ER-negative cell lines. Transfection of ER alpha into ER-negative cell lines restored E2-induced hTERT gene transcription and telomerase activity. Gel shift assays revealed two EREs in the hTERT promoter that specifically bind to ER alpha. Luciferase assays demonstrated that at least the proximal ERE is responsible for transcriptional activation by ligand-stimulated ER alpha.
  
  
  CONCLUSIONS
  Telomerase activity and hTERT mRNA were increased in response to estrogen in an ER alpha-dependent fashion in endometrial cancer cells. Binding of complexed estrogen with ER alpha to the EREs found within the hTERT promoter suggests a possible mechanism for telomerase induction that may facilitate the malignant transformation of hormone-dependent endometrial cells.
authors.affiliation: Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA. jboggess@med.unc.edu
authors.forename: John F
authors.initials: JF
authors.lastname: Boggess
authors.forename: Chunxiao
authors.initials: C
authors.lastname: Zhou
authors.forename: Victoria L
authors.initials: VL
authors.lastname: Bae-Jump
authors.forename: Paola A
authors.initials: PA
authors.lastname: Gehrig
authors.forename: Young E
authors.initials: YE
authors.lastname: Whang
mesh_terms: D045744:Cell Line, Tumor; D016889:Endometrial Neoplasms / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D004958:Estradiol / Q000494:pharmacology; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D005260:Female; D006801:Humans; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D020218:Response Elements; D019098:Telomerase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic; D014162:Transfection
chemical_list: D047628:Estrogen Receptor alpha; D012333:RNA, Messenger; D004958:Estradiol; C509186:TERT protein, human; D019098:Telomerase