title: Analysis of cathepsin D forms and their clinical implications in human prostate cancer.
abstract: OBJECTIVE
  To assess cathepsin D (Cat.D) status in the prostate, we analyzed the different Cat.D forms in human prostate tissues using Western immunoblots.
  
  
  METHODS
  Cell extracts were prepared from prostate tissues (n = 42) obtained from radical prostatectomy, adopting the tissue homogenization method. Expression of the different Cat.D forms was analyzed using Western blots. The catalytic activity of Cat.D was assayed by acid treatment, in which cell extracts were incubated in acidic buffer (pH 3 to 4) at 37C for 1 hour.
  
  
  RESULTS
  Pathologically confirmed normal (NML), benign prostatic hyperplasia (BPH) and cancer (CAP) specimens all expressed Cat.D, but as two distinct forms. Both NML and BPH predominantly expressed an inactive procathepsin D (Pro.Cat.D), while CAP notably exhibited an active mature Cat.D. The assessment of Cat.D activity, using PSA (prostate specific antigen) as a physiological substrate, showed that such activity was consistently higher in CAP than in NML/BPH specimens. Further studies revealed that the mode of Cat.D activation in CAP specimens appeared to be primarily due to acid-induced autoproteolysis (self-degradation) of mature Cat.D.
  
  
  CONCLUSIONS
  This study demonstrates that expression and activity of Cat.D varies among prostate specimens. A greater expression of mature Cat.D with a higher catalytic activity in CAP specimens is the most notable difference from NML/BPH. Therefore, the differential expression/activity of Cat.D forms may be a useful indicator for assessing prostate cancer status.
authors.affiliation: Department of Urology, New York Medical College, Valhalla 10595, USA.
authors.forename: J P
authors.initials: JP
authors.lastname: Cherry
authors.forename: J A
authors.initials: JA
authors.lastname: Mordente
authors.forename: J R
authors.initials: JR
authors.lastname: Chapman
authors.forename: M S
authors.initials: MS
authors.lastname: Choudhury
authors.forename: H
authors.initials: H
authors.lastname: Tazaki
authors.forename: C
authors.initials: C
authors.lastname: Mallouh
authors.forename: S
authors.initials: S
authors.lastname: Konno
mesh_terms: D015153:Blotting, Western; D002402:Cathepsin D / Q000032:analysis* / Q000096:biosynthesis; D006801:Humans; D008297:Male; D011467:Prostate / Q000737:chemistry*; D017430:Prostate-Specific Antigen / Q000032:analysis; D011470:Prostatic Hyperplasia; D011471:Prostatic Neoplasms / Q000737:chemistry*
chemical_list: D017430:Prostate-Specific Antigen; D002402:Cathepsin D