Detection of urothelial bladder cancer cells in voided urine can be improved by a combination of cytology and standardized microsatellite analysis.
OBJECTIVE
To evaluate molecular and immunohistochemical markers to develop a molecular grading of urothelial bladder cancer and to test these markers in voided urine samples.


METHODS
255 consecutive biopsies from primary bladder cancer patients were evaluated on a tissue microarray. The clinical parameters gender, age, adjacent carcinoma in situ, and multifocality were collected. UroVysion fluorescence in situ hybridization (FISH) was done. Expression of cytokeratin 20, MIB1, and TP53 was analyzed by immunohistochemistry. Fibroblast growth factor receptor 3 (FGFR3) status was studied by SNaPshot mutation detection. Results were correlated with clinical outcome by Cox regression analysis. To assess the predictive power of different predictor subsets to detect high grade and tumor invasion, logistic regression models were learned. Additionally, voided urine samples of 119 patients were investigated. After cytologic examination, urine samples were matched with their biopsies and analyzed for loss of heterozygosity (LOH), FGFR3 mutation, polysomy, and p16 deletion using UroVysion FISH. Receiver operator characteristic curves for various predictor subsets were plotted.


RESULTS
In biopsies, high grade and solid growth pattern were independent prognostic factors for overall survival. A model consisting of UroVysion FISH and FGFR3 status (FISH + FGFR3) predicted high grade significantly better compared with a recently proposed molecular grade (MIB1 + FGFR3). In voided urine, the combination of cytology with LOH analysis (CYTO + LOH) reached the highest diagnostic accuracy for the detection of bladder cancer cells and performed better than cytology alone (sensitivity of 88.2% and specificity of 97.1%).


CONCLUSIONS
The combination of cytology with LOH analysis could reduce unpleasant cystoscopies for bladder cancer patients.
Wild
Peter J
PJ
Institute of Surgical Pathology, University Hospital Zurich, Zurich, Switzerland. peter.wild@usz.ch
Fuchs
Thomas
T
Stoehr
Robert
R
Zimmermann
Dieter
D
Frigerio
Simona
S
Padberg
Barbara
B
Steiner
Inbal
I
Zwarthoff
Ellen C
EC
Burger
Maximilian
M
Denzinger
Stefan
S
Hofstaedter
Ferdinand
F
Kristiansen
Glen
G
Hermanns
Thomas
T
Seifert
Hans-Helge
HH
Provenzano
Maurizio
M
Sulser
Tullio
T
Roth
Volker
V
Buhmann
Joachim M
JM
Moch
Holger
H
Hartmann
Arndt
A
D014408:Biomarkers, Tumor / Q000235:genetics* / Q000652:urine*; D003584:Cytological Techniques; D004252:DNA Mutational Analysis; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D017404:In Situ Hybridization, Fluorescence; D053208:Kaplan-Meier Estimate; D019656:Loss of Heterozygosity; D008297:Male; D018895:Microsatellite Repeats / Q000235:genetics*; D051498:Receptor, Fibroblast Growth Factor, Type 3 / Q000096:biosynthesis / Q000235:genetics; D012680:Sensitivity and Specificity; D046888:Tissue Array Analysis; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism; D001749:Urinary Bladder Neoplasms / Q000175:diagnosis / Q000235:genetics* / Q000652:urine*
D014408:Biomarkers, Tumor; C513496:MIB1 ligase, human; D044767:Ubiquitin-Protein Ligases; D051498:Receptor, Fibroblast Growth Factor, Type 3