Katz A E, Olsson C A, Raffo A J, Cama C, Perlman H, Seaman E, O'Toole K M, McMahon D, Benson M C, Buttyan R
Department of Urology, Columbia University College of Physicians and Surgeons, New York, New York.
Urology. 1994 Jun;43(6):765-75. doi: 10.1016/0090-4295(94)90132-5.
Because up to 40 percent of surgically treated patients with prostate cancer are subsequently found to be clinically understaged, a more sensitive staging modality to identify extraprostatic disease prior to surgery is required.
We describe an enhanced reverse transcriptase [RT] polymerase chain reaction (PCR) assay utilizing oligonucleotide primers specific for the human prostate-specific antigen (PSA). This assay identifies PSA-synthesizing cells from reverse transcribed mRNA. This assay was applied to RNAs extracted from the peripheral blood lymphocytes of 65 patients with clinically localized prostate cancer. In addition, blood from 20 women, 20 young men, 25 age-matched control men under treatment for benign prostatic hyperplasia (BPH), and 18 men with established, untreated metastatic prostate cancer was tested.
An RT-PCR assay for PSA can recognize one PSA-expressing cell diluted into one hundred thousand lymphocytes. The sensitivity of this assay can be enhanced by the addition of digoxigenin-modified nucleotides to the PCR reaction and this assay was applied to RNAs extracted from the peripheral lymphocyte fraction of 148 prostate cancer patients and controls at this institution. Although no specimen from women or men without cancer was positive in this assay, 14 of 18 metastatic prostate cancer patients were positive (77.8%). Additionally, 25 of 65 (38.5%) patients with clinically localized disease (T1-2b) were positive from blood specimens obtained prior to surgery. Final pathologic results from this group of patients identified a correlation between positivity on this assay and the presence of capsular tumor penetration (sensitivity, 68%; specificity, 84%) as well as strong correlation with the finding of carcinoma at the surgical margin (sensitivity, 87%; specificity, 76%). Logarithmic regression analysis of the results of the RT-PCR assay indicates its remarkable superiority to digital rectal examination, computed tomography scan, endorectal coil magnetic resonance imaging, PSA, prostate-specific antigen density, or Gleason score for predicting the true pathologic stage of prostate cancer in these surgically treated patients.
An RT-PCR assay using PSA primers to detect prostate cells in the peripheral circulation of surgical-candidate patients is significantly correlated with capsular penetration and tumor-positive surgical margins. This molecular assay provides a sensitive and specific means to stage correctly apparent localized prostate cancer prior to radical prostatectomy.
由于高达40%接受手术治疗的前列腺癌患者随后被发现临床分期过低，因此需要一种更敏感的分期方法，以便在手术前识别前列腺外疾病。
我们描述了一种增强型逆转录[RT]聚合酶链反应（PCR）检测方法，该方法使用针对人前列腺特异性抗原（PSA）的寡核苷酸引物。该检测方法从逆转录的mRNA中识别出合成PSA的细胞。该检测方法应用于65例临床局限性前列腺癌患者外周血淋巴细胞提取的RNA。此外，还检测了20名女性、20名年轻男性、25名年龄匹配的正在接受良性前列腺增生（BPH）治疗的对照男性以及18名已确诊、未经治疗的转移性前列腺癌男性的血液。
PSA的RT-PCR检测方法能够识别稀释在十万个淋巴细胞中的一个表达PSA的细胞。通过在PCR反应中添加地高辛标记的核苷酸可以提高该检测方法的灵敏度，并且该检测方法应用于本机构148例前列腺癌患者和对照的外周淋巴细胞部分提取的RNA。尽管在该检测中没有来自无癌症的女性或男性的标本呈阳性，但18例转移性前列腺癌患者中有14例呈阳性（77.8%）。此外，65例（38.5%）临床局限性疾病（T1-2b）患者中，25例在手术前采集的血液标本呈阳性。这组患者的最终病理结果表明，该检测呈阳性与包膜肿瘤侵犯之间存在相关性（灵敏度为68%；特异性为84%），并且与手术切缘发现癌灶也有很强的相关性（灵敏度为87%；特异性为76%）。对RT-PCR检测结果进行对数回归分析表明，在预测这些接受手术治疗的患者前列腺癌的真实病理分期方面，该检测方法明显优于直肠指检、计算机断层扫描、直肠内线圈磁共振成像、PSA、前列腺特异性抗原密度或Gleason评分。
使用PSA引物的RT-PCR检测方法来检测手术候选患者外周循环中的前列腺细胞，与包膜侵犯和肿瘤阳性手术切缘显著相关。这种分子检测方法为在根治性前列腺切除术之前正确分期明显局限性前列腺癌提供了一种敏感且特异的手段。
