Siracusano Salvatore, Niccolini Benedetta, Knez Roberto, Tiberio Anna, Benedetti Elena, Bonin Serena, Ciciliato Stefano, Pappagallo Giovanni L, Belgrano Emanuele, Stanta Giorgio
Department of Urology, Trieste University, Cattinara Hospital, Via Strada di Fiume 447, 34100 Trieste, Italy.
Eur Urol. 2005 Mar;47(3):327-33. doi: 10.1016/j.eururo.2004.10.007. Epub 2004 Dec 2.
Because of the low sensitivity of urinary cytological diagnosis of urinary bladder carcinoma, new molecular diagnostic methods have been proposed. We decided to verify the expression of telomerase mRNA coding for the catalytic component (hTRT), cytokeratin 20 (CK20) and CD4 antigen mRNAs in urine as possible diagnostic tool.
Evaluation of hTRT, CK20, CD4 mRNAs was performed in 50 ml of naturally voided urine of 205 patients of which 153 with bladder cancer (Tis, n = 11; TaGx, n = 4; TaG1, n = 25; TaG2, n = 26; TaG3, n = 8; T1G1, n = 16; T1G2, n = 17; T1G3, n = 20; T2G2, n = 6; T2G3, n = 13; T3G3, n = 7) and 52 controls. A quantitative expression of hTRT at mRNA level versus TRAP (telomeric repeat amplification protocol) assay was performed in 20 patients and 14 controls. The expression of RT-PCR for hTRT, CK20, CD4 versus urinary cytology was analysed in 44 patients with bladder cancer. Evaluating the three molecular markers together, the result was considered correct when at least two of the markers were positive, suspected when only one marker was positive and negative for diagnosis of tumour when all markers were negative. The performance of the diagnostic model resulted from the logistic analysis evaluated with receiver operating characteristics (ROC) curve analysis.
The sensitivity detected for each tumour marker was as follows: for hTRT 90.8%, for CK20 84.3% and for CD4 was 64.7%, while the specificity was 94.2% for CD4 and 78.8% for both hTRT and CK20. When a simultaneous evaluation of the three tumour markers was considered, 88.2% of the diagnoses were correct, 11.8% were suspected for tumour and none were mistaken. When compared with cytology, the simultaneous use of the three markers allowed reaching a correct diagnosis in 88% of the cases in comparison to 25% by urinary cytology. The sensitivity in the detection of bladder cancer was higher for hTRT at mRNA level in comparison with the enzymatic activity detection with TRAP (90% vs. 35%) while the specificity for both markers resulted very high (100%).
These data show that in the future the diagnostic improvement of urine based molecular markers for the detection of bladder cancer in the urine could improve the sensitivity of urinary cytology reducing the need of a cystoscopy.
由于膀胱癌细胞学诊断的敏感性较低，人们提出了新的分子诊断方法。我们决定验证尿液中编码催化成分的端粒酶mRNA（hTRT）、细胞角蛋白20（CK20）和CD4抗原mRNA的表达，将其作为可能的诊断工具。
对205例患者自然排出的50ml尿液进行hTRT、CK20、CD4 mRNA评估，其中153例为膀胱癌患者（Tis，n = 11；TaGx，n = 4；TaG1，n = 25；TaG2，n = 26；TaG3，n = 8；T1G1，n = 16；T1G2，n = 17；T1G3，n = 20；T2G2，n = 6；T2G3，n = 13；T3G3，n = 7），52例为对照。对20例患者和14例对照进行了mRNA水平的hTRT定量表达与端粒重复序列扩增法（TRAP）检测的比较。分析了44例膀胱癌患者hTRT、CK20、CD4的逆转录聚合酶链反应（RT-PCR）表达与尿液细胞学检查结果的关系。同时评估这三种分子标志物时，当至少两个标志物为阳性时结果被认为正确，仅一个标志物为阳性时结果可疑，所有标志物均为阴性时诊断为肿瘤阴性。诊断模型的性能通过逻辑分析和受试者工作特征（ROC）曲线分析进行评估。
每个肿瘤标志物检测到的敏感性如下：hTRT为90.8%，CK20为84.3%，CD4为64.7%，而CD4的特异性为94.2%，hTRT和CK20均为78.8%。同时评估三种肿瘤标志物时，88.2%的诊断正确，11.8%可疑为肿瘤，无误诊。与细胞学检查相比，同时使用三种标志物在88%的病例中能得出正确诊断（尿液细胞学检查为25%）。与TRAP酶活性检测相比，mRNA水平的hTRT检测膀胱癌的敏感性更高（90%对35%），而两种标志物的特异性都非常高（100%）。
这些数据表明，未来基于尿液的分子标志物用于尿液中膀胱癌检测的诊断改进可能会提高尿液细胞学检查的敏感性，减少膀胱镜检查的需求。
