title: Comparison of telomerase activity and GSTP1 promoter methylation in ejaculate as potential screening tests for prostate cancer.
abstract: New diagnostic tools are needed for the early detection of prostatic cancer. The molecular detection of prostate cancer cells in ejaculates was evaluated using complementary PCR-based methods. LNCaP cells, a cell line derived from prostatic carcinoma, were spiked into normal seminal ejaculates and the prostatic epithelial component of the specimens was isolated by immunomagnetic bead sorting, using a monoclonal antibody to prostate-specific membrane antigen (PSMA). Ejaculates from nine patients with a recent diagnosis of prostate cancer were processed in a similar fashion, using LNCaP-spiked aliquots as an internal positive control. Telomerase expression was evaluated by the telomeric repeat amplification protocol (TRAP) and glutathione S-transferase gene promoter (GSTP1) hypermethylation was evaluated by methylation-sensitive restriction endonuclease digestion and PCR amplification. Telomerase activity was detected in LNCaP cells recovered from normal seminal ejaculates but was not found in all nine samples from patients with prostate cancer. The sensitivity of GSTP1 analysis was similar to telomerase analysis for the detection of LNCaP cells from normal ejaculate samples but was positive in ejaculates from four out of nine patients with prostate cancer. GSTP1 DNA methylation status is more sensitive than telomerase analysis for the detection of malignant cells in seminal ejaculates from patients with prostate cancer.
authors.affiliation: University of Colorado Health Sciences Center and University of Colorado Cancer Center, Denver, CO 80262, USA.
authors.forename: C I
authors.initials: CI
authors.lastname: Suh
authors.forename: T
authors.initials: T
authors.lastname: Shanafelt
authors.forename: D J
authors.initials: DJ
authors.lastname: May
authors.forename: K R
authors.initials: KR
authors.lastname: Shroyer
authors.forename: J B
authors.initials: JB
authors.lastname: Bobak
authors.forename: E D
authors.initials: ED
authors.lastname: Crawford
authors.forename: G J
authors.initials: GJ
authors.lastname: Miller
authors.forename: N
authors.initials: N
authors.lastname: Markham
authors.forename: L M
authors.initials: LM
authors.lastname: Glode
mesh_terms: D019175:DNA Methylation; D004542:Ejaculation; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase / Q000235:genetics*; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D008297:Male; D008403:Mass Screening / Q000379:methods; D011401:Promoter Regions, Genetic; D011471:Prostatic Neoplasms / Q000175:diagnosis* / Q000235:genetics*; D012016:Reference Values; D013094:Spermatozoa / Q000502:physiology; D019098:Telomerase / Q000032:analysis / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D007527:Isoenzymes; C496556:GSTP1 protein, human; D051549:Glutathione S-Transferase pi; D005982:Glutathione Transferase; D019098:Telomerase