title: Ratio of cathepsin B to stefin A identifies heterogeneity within Gleason histologic scores for human prostate cancer.
abstract: BACKGROUND
  Cathepsin B (CB), a lysosomal cysteine protease, is involved in degradation of extracellular matrix proteins and progression of tumor cells from one biological compartment to another in many solid organ cancers, including prostate cancer. Our objective was to identify patterns of distribution of CB and its endogenous cellular inhibitor stefin A in cryostat sections of frozen BPH and prostate cancer tissue samples and to define these patterns in relation to Gleason histologic scores, clinical stages, and serum total PSA levels.
  
  
  METHODS
  We localized CB and stefin A in the same sections using polyclonal and monoclonal antibody immunoglobulin G (IgGs) against CB and stefin A using immunofluorescence and confocal microscopic techniques. Only cryostat sections of frozen prostates were used in localizations of CB and stefin A.
  
  
  RESULTS
  Benign prostatic hyperplasia (BPH) showed similar localization patterns for CB and stefin A and a ratio of 1 was indicated by CB = stefin A. Confocal studies indicated that most CB and stefin A sites in BPH glandular cells overlapped as shown by the yellow fluorescence of their co-localization. We found considerable variability in individual localization of CB and stefin A within and between Gleason histologic scores for prostate cancers. This variability was also found in Gleason score 6 tumors that are otherwise considered similar histologically and morphologically. Negative control sections did not show localization of CB by FITC, stefin A by Cy3 or yellow fluorescence for co-localization. Our analysis of the ratio of CB to stefin A showed three patterns, namely CB = stefin A, CB > stefin A, and CB < stefin A, within each Gleason score evaluated by us. Confocal microscopy showed more sites of yellow fluorescence when the ratio was CB = stefin A than those found in CB > stefin A or CB < stefin A. Statistical analyses showed prostate cancer cases with ratios of CB > stefin A (P < 0.05) and CB < stefin A (P < 0.05) significantly different from normal prostate and BPH which had ratios of CB = stefin A. Regression analysis did not show any specific relationship between the ratio of CB to stefin A and Gleason scores, clinical stages, and serum total prostate specific antigen (PSA) levels in prostate cancers. Analysis of our data indicates that the homeostatic balance between the enzyme and inhibitor was altered even in Gleason histologic score 6 tumors that are usually considered histologically similar by glandular differentiation.
  
  
  CONCLUSIONS
  We have shown that prostate cancer is a heterogeneous tumor within each Gleason histological score regardless of the progression indicated by lower to higher Gleason score tumors. The ratio of CB > stefin A would indicate a preponderance of enzyme that would favor degradation of extracellular matrix proteins and progression of tumor cells in biological compartments. These tumors are expected to be aggressive prostate cancers. In contrast, prostate tumors showing ratios of CB < stefin A and CB = stefin A are expected to be less aggressive prostate cancers. This is the first report to define heterogeneity within any Gleason score for prostate cancers by the ratios of CB to stefin A.
authors.affiliation: Department of Genetics, Cell Biology & Development, University of Minnesota, Minneapolis, Minnesota, USA. sinha001@tc.umn.edu
authors.forename: A A
authors.initials: AA
authors.lastname: Sinha
authors.forename: B J
authors.initials: BJ
authors.lastname: Quast
authors.forename: M J
authors.initials: MJ
authors.lastname: Wilson
authors.forename: E T
authors.initials: ET
authors.lastname: Fernandes
authors.forename: P K
authors.initials: PK
authors.lastname: Reddy
authors.forename: S L
authors.initials: SL
authors.lastname: Ewing
authors.forename: B F
authors.initials: BF
authors.lastname: Sloane
authors.forename: D F
authors.initials: DF
authors.lastname: Gleason
mesh_terms: D000368:Aged; D000369:Aged, 80 and over; D002401:Cathepsin B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015891:Cystatins / Q000378:metabolism*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008297:Male; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D008875:Middle Aged; D017430:Prostate-Specific Antigen / Q000097:blood; D011470:Prostatic Hyperplasia / Q000201:enzymology* / Q000473:pathology; D011471:Prostatic Neoplasms / Q000201:enzymology* / Q000473:pathology*; D012044:Regression Analysis
chemical_list: D015891:Cystatins; D017430:Prostate-Specific Antigen; D002401:Cathepsin B