Proteome analysis of gelatin-bound urinary proteins from patients with bladder cancers.
OBJECTIVE
Proteome analysis with protein separation by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) and mass spectrometric measurement is utilized for comprehensive identification of proteins in various biological samples. However, this technique has not been widely applied for the analysis of body fluid for clinical assessments, because large amounts of proteins in fluid such as albumin and globulin hinder the detection of small amounts of proteins with high clinical values. The aim of the present study is to develop a method of proteomic analysis for urine of patients with bladder cancers.


METHODS
Urine samples collected from 40 patients with bladder cancers, 32 healthy volunteers, and 7 old volunteers with benign prostate hypertrophy were treated with gelatin-beads as a group-specific affinity carrier. The gelatin-bound proteins were subjected to the proteome analysis.


RESULTS
Various amounts of matrix metalloproteinase-2 (MMP-2), -9 (MMP-9), fibronectin (FN), and their fragments were identified on 2-D PAGE gels from cancer-bearing patients, but not from normal individuals. Importantly, not only total amounts of these proteins, but also patterns of their distributions on the gels were well matched with the extent of invasion diagnosed with histopathological examinations. Thus, the score including such qualitative patterning of these proteins predicts pathological stages of the cancer.


CONCLUSIONS
The proteomic analysis with gelatin-affinity purification of urine samples is useful not only for the diagnosis of bladder cancers, but also for estimating the extent of tumor invasion.
Saito
Minoru
M
Fukuyama Medical Laboratory, Fukuyama, Japan.
Kimoto
Masafumi
M
Araki
Teruo
T
Shimada
Yukie
Y
Fujii
Ryoji
R
Oofusa
Ken
K
Hide
Michihiro
M
Usui
Tsuguru
T
Yoshizato
Katsutoshi
K
D014408:Biomarkers, Tumor / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005353:Fibronectins / Q000032:analysis; D005780:Gelatin / Q000737:chemistry*; D006801:Humans; D020778:Matrix Metalloproteinase 2 / Q000032:analysis; D020780:Matrix Metalloproteinase 9 / Q000032:analysis; D010446:Peptide Fragments / Q000032:analysis; D011237:Predictive Value of Tests; D011506:Proteins / Q000032:analysis*; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D001749:Urinary Bladder Neoplasms / Q000175:diagnosis / Q000378:metabolism; D014556:Urine / Q000737:chemistry*
D014408:Biomarkers, Tumor; D005353:Fibronectins; D010446:Peptide Fragments; D011506:Proteins; D020543:Proteome; D005780:Gelatin; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9