title: Improved TRAP-silver staining versus conventional radioactive TRAP assays: quantification of telomerase activity during immortalization and in pathological human endometrium.
abstract: OBJECTIVE
  To develop a sensitive telomeric repeat amplification protocol (TRAP)-silver staining assay for telomerase activity quantification.
  
  
  METHODS
  TRAP assays were performed by using a TRAPeze telomerase kit with or without [alpha-32P]-dCTP. Amplification products were electrophoresed in polyacrylamide gels and detected by autoradiography or a modified silver staining protocol. Telomerase activity was quantified from radioactive counts or optical density of telomerase products from test extracts and controls.
  
  
  RESULTS
  TRAP-silver staining assay was at least as sensitive as radioactive TRAP assay and quantified telomerase activity within linearity from 10 to 3,000 cell equivalents. Both methods quantified a weak telomerase activity in normal endometrial glandular epithelial cells (GEC) and a strong increase in immortalized GEC. In human pathologic endometria (n=24), telomerase activity was correlated with lesion seriousness and distinguished simple hyperplasias from nonhyperplasic or cancerous lesions.
  
  
  CONCLUSIONS
  TRAP-silver staining assay is suitable for cell and tissue telomerase activity routine quantification.
authors.affiliation: Laboratoire de Biochimie Biologie Moléculaire, Université de Franche-Comté, UFR Sciences et Techniques, Besançon, France.
authors.forename: C
authors.initials: C
authors.lastname: Nemos
authors.forename: J P
authors.initials: JP
authors.lastname: Rémy-Martin
authors.forename: P
authors.initials: P
authors.lastname: Adami
authors.forename: F
authors.initials: F
authors.lastname: Arbez-Gindre
authors.forename: J P
authors.initials: JP
authors.lastname: Schaal
authors.forename: M
authors.initials: M
authors.lastname: Jouvenot
authors.forename: R
authors.initials: R
authors.lastname: Delage-Mourroux
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D004717:Endometrium / Q000737:chemistry* / Q000201:enzymology* / Q000473:pathology; D005260:Female; D006168:Guinea Pigs; D006367:HeLa Cells; D006801:Humans; D008875:Middle Aged; D016622:Silver Staining / Q000379:methods*; D019098:Telomerase / Q000032:analysis* / Q000378:metabolism*
chemical_list: D019098:Telomerase