Corey E, Arfman E W, Oswin M M, Melchior S W, Tindall D J, Young C Y, Ellis W J, Vessella R L
Urology Department, School of Medicine, University of Washington, Seattle 98195, USA.
Urology. 1997 Aug;50(2):184-8. doi: 10.1016/S0090-4295(97)00262-8.
To investigate the clinical value of human glandular kallikrein (hK2) reverse transcriptase-polymerase chain reaction (RT-PCR) for detection of prostate cells in circulation and to compare the results with those obtained from prostate-specific antigen (PSA) RT-PCR.
We examined peripheral blood (PB) and bone marrow (BM) samples of 13 patients with advanced-stage prostate cancer and 63 patients with clinically localized disease for the presence of circulating prostate cells. An RT-PCR protocol with a two-step amplification cycle and hot-start conditions was used.
The limit of detection of the PCR portion is similar for PSA and hK2 (5 to 10 copies of the plasmid containing the cDNA). The RT-PCR limit of detection is one LNCaP cell in 10(8) peripheral blood mononuclear cells (PMBC) for PSA, and one LNCaP cell in 10(7) PMBC for hK2. Of the BM samples obtained prior to radical prostatectomy, 71.4% were positive for PSA mRNA and 41.3% were positive for hK2 mRNA. In PB, the PSA positivity was 19% and hK2 positivity 12.7%. In advanced-stage patients, there were 76.9% PSA-positive samples in BM versus 38.5% hK2-positive samples; 46.2% of patients were positive in PB for PSA versus 30.8% for hK2.
We have developed a sensitive RT-PCR protocol for detection of hK2 mRNA and evaluated the suitability of hK2 mRNA in comparison with PSA mRNA as an additional marker for detection of prostate cells in circulation. Combining results of these two tests increased the sensitivity of detection.
研究人腺激肽释放酶（hK2）逆转录聚合酶链反应（RT-PCR）检测循环中前列腺细胞的临床价值，并将结果与前列腺特异性抗原（PSA）RT-PCR的结果进行比较。
我们检测了13例晚期前列腺癌患者和63例临床局限性疾病患者的外周血（PB）和骨髓（BM）样本中循环前列腺细胞的存在情况。使用了具有两步扩增循环和热启动条件的RT-PCR方案。
PCR部分的检测限对于PSA和hK2相似（含有cDNA的质粒5至10个拷贝）。RT-PCR检测限对于PSA是10⁸外周血单个核细胞（PMBC）中有1个LNCaP细胞，对于hK2是10⁷ PMBC中有1个LNCaP细胞。在前列腺癌根治术前获取的BM样本中，71.4%的样本PSA mRNA呈阳性，41.3%的样本hK2 mRNA呈阳性。在PB中，PSA阳性率为19%，hK2阳性率为12.7%。在晚期患者中，BM中PSA阳性样本为76.9%，而hK2阳性样本为38.5%；PB中PSA阳性患者为46.2%，而hK2阳性患者为30.8%。
我们开发了一种用于检测hK2 mRNA的灵敏RT-PCR方案，并评估了hK2 mRNA与PSA mRNA相比作为检测循环中前列腺细胞的附加标志物的适用性。结合这两项检测结果提高了检测的敏感性。
