Molecular profiling of ADAM12 in human bladder cancer.
OBJECTIVE
We have previously found ADAM12, a disintegrin and metalloprotease, to be an interesting biomarker for breast cancer. The purpose of this study was to determine the gene and protein expression profiles of ADAM12 in different grades and stages of bladder cancer.


METHODS
ADAM12 gene expression was evaluated in tumors from 96 patients with bladder cancer using a customized Affymetrix GeneChip. Gene expression in bladder cancer was validated using reverse transcription-PCR, quantitative PCR, and in situ hybridization. Protein expression was evaluated by immunohistochemical staining on tissue arrays of bladder cancers. The presence and relative amount of ADAM12 in the urine of cancer patients were determined by Western blotting and densitometric measurements, respectively.


RESULTS
ADAM12 mRNA expression was significantly up-regulated in bladder cancer, as determined by microarray analysis, and the level of ADAM12 mRNA correlated with disease stage. Reverse transcription-PCR, quantitative PCR, and in situ hybridization validated the gene expression results. Using immunohistochemistry, we found ADAM12 protein expression correlated with tumor stage and grade. Finally, ADAM12 could be detected in the urine by Western blotting; ADAM12 was present in higher levels in the urine from patients with bladder cancer compared with urine from healthy individuals. Significantly, following removal of tumor by surgery, in most bladder cancer cases examined, the level of ADAM12 in the urine decreased and, upon recurrence of tumor, increased.


CONCLUSIONS
ADAM12 is a promising biomarker of bladder cancer.
Fröhlich
Camilla
C
Institute of Molecular Pathology, University of Copenhagen, Copenhagen, Denmark.
Albrechtsen
Reidar
R
Dyrskjøt
Lars
L
Rudkjaer
Lise
L
Ørntoft
Torben F
TF
Wewer
Ulla M
UM
D051722:ADAM Proteins / Q000378:metabolism* / Q000652:urine; D000072197:ADAM10 Protein; D000072199:ADAM12 Protein; D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology / Q000601:surgery / Q000652:urine; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D053829:Amyloid Precursor Protein Secretases / Q000378:metabolism; D000818:Animals; D002294:Carcinoma, Squamous Cell / Q000378:metabolism / Q000473:pathology / Q000601:surgery / Q000652:urine; D002295:Carcinoma, Transitional Cell / Q000378:metabolism* / Q000473:pathology / Q000601:surgery / Q000652:urine; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D008325:Mammary Neoplasms, Experimental / Q000378:metabolism / Q000473:pathology; D008565:Membrane Proteins / Q000378:metabolism* / Q000652:urine; D051379:Mice; D008875:Middle Aged; D009092:Mucous Membrane / Q000378:metabolism; D009364:Neoplasm Recurrence, Local / Q000652:urine; D009367:Neoplasm Staging; D001749:Urinary Bladder Neoplasms / Q000378:metabolism* / Q000473:pathology / Q000601:surgery / Q000652:urine
D008565:Membrane Proteins; D053829:Amyloid Precursor Protein Secretases; D051722:ADAM Proteins; D000072199:ADAM12 Protein; C000606480:ADAM12 protein, human; C068490:ADAM8 protein, human; D000072197:ADAM10 Protein; C491236:ADAM10 protein, human