Proliferating cell nuclear antigen in normal urothelium and urothelial lesions of the urinary bladder: a quantitative assessment using a true color image analysis system |
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Authors: | Ruth Knuechel Markus Burgau Josef Rueschoff Ferdinand Hofstaedter |
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Institution: | 1. Institute of Pathology, University of Regensburg, Franz-Josef-Strauss Allee 11, W-8400, Regensburg, Germany
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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. In contrast with data from
normal tissue and malignant hematological neoplasms, the amount of PCNA is regulated differently in urothelial neoplasms,
emphasizing the biological differences between the following two sets: mild dysplasia and moderate dysplasia; mild dysplasia
and papillary carcinomas. The use of image analysis to standardize the detection process after controlled staining conditions
is advisable in order to provide reliable data.
Supported by the DFG project: Knuechel/Urothelcarcinom 263 |
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Keywords: | Proliferating cell nuclear antigen (PCNA) Immunohistochemistry Image analysis Urothelial lesions Bladder cancer cell line |
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