DNA-based detection of prostate cancer in blood, urine, and ejaculates.
BACKGROUND
Methylation-specific polymerase chain reaction (MSP) targeting promoter hypermethylation of the glutathione S transferase P1 gene (GSTP1), as the most frequent DNA alteration in prostatic carcinoma, was used for the molecular detection of cell-bound and cell-free prostate tumor DNA in various human bodily fluids.


METHODS
We investigated GSTP1 promoter hypermethylation in DNA isolated from plasma, serum, nucleated blood cells, ejaculates, urines after prostate massage, and prostate tissue from 33 patients with prostate cancer and 26 control patients with benign prostatic hyperplasia (BPH). Using a viral DNA extraction kit specifically recommended for DNA isolation from urine samples, GSTP1 promoter hypermethylation in urine sediments after prostatic massage was investigated in a cohort of 29 patients with prostate cancer and 40 controls with BPH. Fluorescently labeled MSP products were analyzed on an automated gene sequencer.


RESULTS
GSTP1 promoter hypermethylation was found in 90% of tumors (18 of 20), 72% of plasma or serum samples (23 of 32), 50% of ejaculates (4 of 8), and between 36% (4 of 11; normal DNA isolation kit) and 76% (22 of 29; viral kit) of exprimated urines from patients with prostate cancer. Also, MSP identified circulating tumor cells in 30% (10 out of 33) of prostate cancer patients. Except for one urine sample, GSTP1 promoter hypermethylation was not found in tissue and body fluids from patients with BPH.


CONCLUSIONS
GSTP1 promoter methylation analysis provides a highly specific tool for DNA-based diagnosis of prostate cancer in body fluids.
Goessl
C
C
Department of Urology, Klinikum Benjamin Franklin, Freie Universität Berlin, Germany. goessl@zedat.fu-berlin.de
Müller
M
M
Heicappell
R
R
Krause
H
H
Miller
K
K
D000368:Aged; D001483:Base Sequence; D016022:Case-Control Studies; D019175:DNA Methylation; D017931:DNA Primers; D004273:DNA, Neoplasm / Q000032:analysis* / Q000097:blood / Q000652:urine; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D008297:Male; D008875:Middle Aged; D011471:Prostatic Neoplasms / Q000097:blood / Q000175:diagnosis* / Q000235:genetics / Q000652:urine; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004273:DNA, Neoplasm; D005982:Glutathione Transferase