title: Proliferating cell nuclear antigen in normal urothelium and urothelial lesions of the urinary bladder: a quantitative assessment using a true color image analysis system.
abstract: To evaluate proliferating cell nuclear antigen (PCNA) staining for assessing proliferative activity in routine pathology specimens of urinary bladder, the bladder carcinoma cell line J82 and a total of 122 specimens of normal bladder and urothelial lesions were stained with the antibody clone PC10 against proliferating cell nuclear antigen. In in vitro plateau cultures the proportion of PCNA-positive cells exceeded that of Ki-67-positive cells, and only very few cells were negative. In formalin-fixed tissues, the PCNA staining pattern, which should be confined to replicon units in the nucleus, was optimized by 1 h postfixation in an organic solvent (methacarn). Sections showed positive nuclear staining confined to basal and some suprabasal cells in normal urothelium and grade 1 dysplasias, but more generalized nuclear staining in all other neoplastic lesions. In addition, stromal cells adjacent to invasive tumors showed nuclear positivity in some instances. Using quantitative true color image analysis of sections counterstained with hemalum, the degree of brown staining of the PCNA reaction product is contrasted with the blue staining of the nuclear area. With this method low contrast specific staining not appreciated optically can be reliably detected. Image analysis data confirmed observations made on noncounterstained sections and showed significant differences between grade 1 and 2 dysplasias as well as between grade 1 dysplasia and all grades of papillary tumor. Furthermore, a significant difference in PCNA staining indices was found between grade 1 and 3 bladder carcinomas. The results indicate that PCNA staining using the PC10 antibody is not confined to the proliferative fraction of neoplastic urothelium.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Institute of Pathology, University of Regensburg, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Knuechel
authors.forename: M
authors.initials: M
authors.lastname: Burgau
authors.forename: J
authors.initials: J
authors.lastname: Rueschoff
authors.forename: F
authors.initials: F
authors.lastname: Hofstaedter
mesh_terms: D000951:Antigens, Neoplasm / Q000032:analysis*; D002295:Carcinoma, Transitional Cell; D004848:Epithelium / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D009687:Nuclear Proteins / Q000032:analysis*; D018809:Proliferating Cell Nuclear Antigen; D014407:Tumor Cells, Cultured; D001743:Urinary Bladder / Q000276:immunology* / Q000473:pathology; D001749:Urinary Bladder Neoplasms / Q000276:immunology* / Q000473:pathology
chemical_list: D000951:Antigens, Neoplasm; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen