Kinetic fluorescence reverse transcriptase-polymerase chain reaction for alpha-methylacyl CoA racemase distinguishes prostate cancer from benign lesions.
BACKGROUND
High-throughput gene expression profiling has recently shown that the mRNA for alpha-methylacyl CoA racemase (AMACR or P504S) is overexpressed in prostate carcinomas (PCa). Several immunohistochemical studies have reported the usefulness of anti-AMACR/P504S for detecting prostate cancer over the full range of prostate specimens encountered in surgical pathology. We tested real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for specific and sensitive detection of AMACR transcripts as a supplementary measure for discriminating between malignant and benign lesions in prostatic tissues.


METHODS
Total RNA was isolated from snap-frozen chips in 55 cases of benign prostatic hyperplasia (BPH) and from frozen sections in 57 prostatectomy cases. The latter were analyzed by an uropathologist (J.K.) and found to contain at least 50% malignant epithelia. Relative quantification of AMACR transcripts was performed by RT-PCR using hybridization probes for detection and PBGD for normalization.


RESULTS
Normalized AMACR transcript levels showed an average 3.75-fold increase in 57 prostate carcinomas cases when compared to 55 cases of BPH (p<0.0001). A 85.6% specificity and 64.9% sensitivity can be achieved if the cutoff is set at 12.95. AMACR expression levels among PCa cases were not statistically associated with the tumor and lymph node stage, the grading, the surgical margins, the Gleason score or progression.


CONCLUSIONS
The present study demonstrates the usefulness of quantitative AMACR-mRNA transcript detection in prostatic tissues as an alternative to immunological staining techniques. Since the latter clearly predominate in the laboratory routine, PCR-based detection of AMACR has the potential to gain widespread acceptance as a suitable future tool for monitoring prostate cancer patients.
Schostak
Martin
M
Department of Urology, Charité-Campus Benjamin Franklin, Universitätsmedizin Berlin, Germany. martin.schostak@charite.de
Miller
Kurt
K
Krause
Hans
H
Schrader
Mark
M
Kempkensteffen
Carsten
C
Kollermann
Jens
J
D000230:Adenocarcinoma / Q000175:diagnosis / Q000201:enzymology / Q000556:secondary; D014408:Biomarkers, Tumor; D005453:Fluorescence; D006801:Humans; D007700:Kinetics; D008297:Male; D011468:Prostatectomy; D011470:Prostatic Hyperplasia / Q000175:diagnosis* / Q000201:enzymology / Q000235:genetics; D011471:Prostatic Neoplasms / Q000175:diagnosis* / Q000201:enzymology / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000235:genetics / Q000378:metabolism; D012372:ROC Curve; D019998:Racemases and Epimerases / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity
D014408:Biomarkers, Tumor; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D019998:Racemases and Epimerases; C108352:alpha-methylacyl-CoA racemase