Non-invasive molecular detection of bladder cancer recurrence

Transitional cell carcinoma (TCC) is the most common bladder tumor and approximately 90% of bladder TCC are superficial at initial diagnosis. High recurrence rate and possible progression to muscle invasive disease that is eventually indicated for radical cystectomy are established features of these tumors. Therefore, reliable predictors of tumor recurrence are of critical importance for management of superficial bladder TCC. Successful molecular diagnosis of bladder cancer by detecting genetic lesions: loss of heterozygosity (LOH) or microsatellite instability (MSI) in cells exfoliated in urine has been reported by several groups including ours. The aim of our study was to evaluate the predictive potential of microsatellite analysis of cells exfoliated in urine in the detection of superficial bladder TCC recurrence. We studied 47 Caucasian patients with confirmed superficial bladder TCC (37 pTa, 10 pT1) at initial diagnosis. Blood samples were obtained once from every patient whereas urine samples were collected before each cystoscopy (initial and follow-up). Matched DNAs from blood and urine were subjected to microsatellite analysis in a blinded fashion. The follow-up period ranged 12-48 months after tumor resection. Microsatellite analysis correctly identified 94% (44/47) of primary tumors and 92% (12/13) of tumor recurrences. Interestingly enough, 75% (9/12) of tumor recurrences were molecularly detected 1-9 months before cystoscopic evidence of recurrent disease. This study demonstrated clearly that not only urine microsatellite analysis reliably detected superficial bladder tumors, but also was a reliable test for detecting and predicting tumor recurrence in Caucasian patients. These results warrant multicenter randomized trials.     

Sensitive detection of transitional cell carcinoma of the bladder by microsatellite analysis of cells exfoliated in urine.

Transitional cell carcinoma (TCC) is the most common bladder tumor. Urine cytology can identify most high-grade tumors but sensitivity is lower if one includes lesions of all grades. Microsatellite marker alterations have been found in many tumor types including bladder cancer and have been used to detect cancer cells in body fluids including urine. The aim of our study is to further evaluate feasibility and sensitivity of microsatellite analysis to detect bladder cancer cells in urine. We studied 55 individuals: 21 with symptoms suggestive of bladder cancer, 23 patients with previous history of TCC and 11 healthy subjects. Genomic DNA was extracted from blood lymphocytes, urine sediment, bladder washings and tumor or normal bladder mucosa. Twenty highly informative microsatellite markers were analyzed for loss of heterozigosity (LOH) and microsatellite instability (MIN) by polymerase chain reaction. Microsatellite analysis of urine identified 33 of 34 (97%) patients with either primary or tumor recurrence, whereas urine cytology identified 27 of 34 (79%) patients (p = 0.0001). Detection of microsatellite abnormalities improved the sensitivity of detecting low-grade and/or stage bladder tumor: from 75-95% for grades G1-G2 and from 75-100% for pTis-pTa tumors. Bladder washings from 25 patients were also analyzed, and in all cases results were identical to those obtained from voided urine. None of the 16 patients without evidence of TCC showed LOH and/or MIN in urine samples or bladder washings. Interestingly, in a patient with persistent bladder mucosa abnormalities, microsatellite alterations were demonstrated 8 months before the histopathologic diagnosis of tumor recurrence. These results further indicate that microsatellite marker analysis is more sensitive than conventional urine cytology in detecting bladder cancer cells in urine and represents a potential clinical tool for monitoring patients with low-grade/stage TCC.     

Microsatellite alterations in urinary sediments from patients with cystitis and bladder cancer

Microsatellite instability (MIN) and loss of heterozygosity (LOH) in bladder cancer have been suggested for diagnosis and follow-up of bladder cancer based on urinary sediments, reflecting tumor alterations. We have examined 6 microsatellites in urine sediments from 11 patients with transitional-cell carcinomas (TCC) and in 31 patients with benign prostatic hyperplasia (BPH), 22 of whom had cystitis. In the TCC patients, tumor tissue was available for comparison with urine. Microsatellites were amplified by PCR and compared with leukocyte DNA from the same individual in silver-stained gels. Altered mobility of bands, new bands and loss of bands were scored. We found MIN and LOH at relatively high frequency in markers from chromosomes 8 and 14 in urine from patients with TCC, but also in BPH patients who had cystitis. Even control patients with BPH without cystitis showed some instability and some losses. Novel bands in urine occurred significantly more often among TCC patients than among BPH patients with or without cystitis (p < 0.001). Band shifts in urine appeared to be more associated with BPH plus cystitis than with TCC. The alterations we found in urine from patients with bladder cancer did not always reflect those found in their tumors, the occurrence of novel bands being significantly higher (p < 0.008) in tumor tissue than in corresponding urine. In conclusion, microsatellite alterations in urine are indicators not only of malignancy but also of inflammatory conditions.     

应用荧光原位杂交技术检测膀胱癌的研究

目的应用荧光原位杂交((fluorescence in situ hybridization,FISH)技术检测膀胱癌患者尿液脱落细胞中染色体异常,评估FISH在中国人群中诊断膀胱癌的作用。方法2007年1月至2008年8月,随机留取20例良性前列腺增生症患者的新鲜尿液,用3号和7号、17号及p16位两组混合探针,通过在尿液脱落细胞标本上进行FISH检测,建立正常人群的阈值;其后随机留取30例门诊膀胱镜活检证实的膀胱癌患者的尿液,同时进行尿液脱落细胞的细胞形态学分析及FISH检测,对比检查结果。结果3号、7号和17号染色体非整倍性改变及p16位点异常正常阈值分别为8.5%、7.1%、6.8%和9.2%,FISH与细胞学检查总敏感性分别为76.6%和43.3%(P<0.05)。T_(is)及T_a、T_1患者FISH检测的敏感性分别为80.0%和64.2%,脱落细胞组织学检测显示敏感性分别为40.0%和35.7%;T_(2-3)患者FISH的敏感性为90.9%,而脱落细胞组织学检测为54.7%(P<0.05),低级别尿路上皮癌FISH及细胞学敏感性分别为68.4%和31.6%;高级别分别为90.9%和63.6%。结论与尿液脱落细胞组织学检测相比,对尿液脱落细胞进行FISH检测可以提高膀胱癌的诊断率,FISH可以作为诊断膀胱癌的一种无创伤的新方法。     

Microsatellite analysis--DNA test in urine competes with cystoscopy in follow-up of superficial bladder carcinoma: a phase II trial

BACKGROUND: It has been shown that microsatellite analysis (MA) is able to detect bladder carcinoma in urine. Relatively small groups of patients often with high stage and grade disease were investigated. However, greater than 85% of cystoscopies are performed for follow-up of superficial bladder carcinoma. The authors evaluated this DNA-based method in a group of consecutive patients in follow-up after transurethral resection of superficial disease. METHODS: Matched blood and urine samples from 109 patients were obtained before cystoscopy and subjected to MA. The BTA stat test (Bard Diagnostic Sciences, Inc., Redmond, WA) and cytology were used for comparison. RESULTS: Sixteen patients were excluded: the DNA was of insufficient quality for 7 patients and leukocyte abundance rendered the result of MA unreliable for 9 patients. For the remaining 93 patients, MA detected 18 of the 24 recurrent tumors. The six undetected tumors were small pTaG1 lesions for which immediate surgery was not necessary. Conversely, 5 of 9 patients with a positive MA and a negative cystoscopy had a tumor recurrence within 6 months after urine collection. In contrast, a recurrence occurred in only 7 of 60 patients who were negative in both MA and cystoscopy (P = 0.006). The MA (74%) appeared more sensitive than the BTA stat test (56%) or urine cytology (22%). CONCLUSIONS: Microsatellite analysis is a DNA test in urine that reliably signals the presence of recurrent bladder carcinoma, sometimes even before cystoscopic evidence of the disease. This noninvasive diagnostic tool has the potential to replace cystoscopy in many cases. The authors' results warrant the need for randomized trials.     

Urinary cytokeratin 20 mRNA expression has the potential to predict recurrence in superficial transitional cell carcinoma of the bladder

Higher levels of cytokeratin 20 (CK 20) mRNA are expressed in malignant urothelial tissue compared to normal tissue. We determined the CK 20 mRNA expression in urine from patients with transitional cell carcinoma (TCC) of the bladder and assessed the biological behavior of such tumors in a 5-year follow-up. Second voided urine was preoperatively collected from 56 patients with bladder carcinoma, from 20 patients with nonmalignant urological diseases and from 40 healthy volunteers. RNA extraction from exfoliated urothelial cells was followed by quantitative real-time RT-PCR with the Light Cycler. Patients in the superficial TCC group had a median expression of 8226AU (arbitrary units) with and 1523AU without tumor recurrence (P=0.023). No such correlation was detected in the group with muscle-invasive tumors. Kaplan-Meier analysis revealed a significant difference between recurrent and nonrecurrent disease (P=0.019) in superficial but not in muscle-invasive TCC (P=0.84). CK 20 mRNA expression in urine has the potential to identify patients at risk for recurrence of noninvasive papillary urothelial tumors. It helps to categorize patients prior to TUR-B, so that the cystoscopy interval during follow-up may be extended in those with low-risk superficial TCC.     

Clinical significance of urinary vascular endothelial growth factor in patients with superficial bladder tumors

Vascular endothelial growth factor (VEGF) is an important mediator of tumor angiogenesis and has been shown to be excreted in the urine of bladder cancer patient. The goal of this study was to evaluate urinary VEGF levels of patients with superficial bladder transitional cell carcinoma (TCC) and to determine its predictive value for recurrence. Pre-operative urinary VEGF levels were determined in 31 patients with superficial bladder TCC and 10 control patients. A quantitative enzyme immunoassay was used to measure urinary VEGF levels and the urine VEGF concentration was corrected by the creatinine concentration in a 24-h urine specimen. The corrected urinary VEGF levels were higher in patients than controls (p=0.003). Ten of 31 patients had TCC recurrences during this study. Corrected urinary VEGF levels were significantly higher in recurrent vs. non-recurrent patients (p=0.001). A cut-off value of 0.32 (corrected urinary VEGF levels) was valuable for predicting recurrences in this prospective study. However, there was no statistical correlation between VEGF levels and tumor stage (Ta or T1), tumor size or tumor grade. Pre-operative urinary VEGF levels are associated with a risk of recurrence in patient with superficial bladder TCC. Quantification of urinary VEGF may prove to be a valuable, non-invasive indicator of carcinoma recurrence in patients with superficial bladder TCC. Urinary VEGF may be a therapeutic target for intravesical therapy. However, because of the small number of cases, further studies with larger number of patients will be needed to clarify this issue.     

Early diagnosis and monitoring of superficial transitional cell carcinoma by microsatellite analysis on urine sediment

It has recently been shown that allelic abnormalities, detected by microsatellite analysis of the DNA extracted from urine sediment, can be successfully used for the detection of transitional cell carcinoma (TCC) of the bladder. The diagnostic accuracy of urinary cytology, urinary bladder cancer (UBC) marker, bladder tumor antigen (BTA) and microsatellite sequence alterations was compared in 42 patients who were recruited for the study. Of them, 30 had been diagnosed with TCC at cystoscopy plus biopsy (group A). Seven patients without any apparent lesions after trans-urethral resection (TUR) and 6 subsequent weeks of endovesical administration of bacillus Calmette-Guerin (BCG), had irritative symptoms. None of them had positive cytology or TCC bladder mucosa biopsies (group B). In the control group were 5 other subjects who were affected by benign prostatic hypertrophy and candidates for prostatectomy (group C). Urine and blood samples were obtained from all of the patients before surgery. Tumor tissue and normal mucosa samples were taken from groups A and C during surgery. Different urinary sediment analyses were performed by using both nuclear medicine and molecular tests. UBC and BTA-t analyses were carried out using monoclonal antibody tests while microsatellite analyses were performed using extracted DNA and electrophoresis of polymerase chain reaction (PCR) products on 13 different primers. Urinary cytological examinations were carried out using the Autocyte Preparation System(R). Urinary cytology confirmed the presence of TCC in 13.3% of patients. The BTA-t marker allowed the identification of 73.3% of cancers with 50% specificity; the UBC marker identified 63.3% of the cases with 41.6% specificity. Microsatellite analysis permitted the identification of 83.3% of the tumors with 100% specificity. DNA analysis demonstrated high sensitivity in patients affected by superficial (81.4%) or G1 (80%) tumors, even when cytological studies demonstrated little or no sensitivity. Microsatellite analysis is a highly-sensitive and specific marker for TCC diagnosis and its monitoring, especially in patients with low-stage and low-grade tumors. Other testing procedures failed to increase urinary cytological diagnostic significance.     

Detecting bladder cancer in the Chinese by microsatellite analysis: ethnic and etiologic considerations

BACKGROUND: Microsatellite analysis of urine sediments has shown promise as a highly sensitive and specific technique for the detection of bladder cancer. However, most studies have been conducted in Western countries with Caucasian subjects. We explored the potential of microsatellite analysis for detecting bladder cancer in Chinese people. METHODS: We performed microsatellite analysis of surgical specimens and urine sediment cells collected from Chinese patients with bladder tumors. Those microsatellite markers giving clearly readable patterns and showing susceptibility to alterations were used as a panel to detect primary tumors. A blinded study of additional patients with bladder cancer was performed to investigate the practical value of this panel for detecting bladder cancer. All statistical tests were two-sided. RESULTS: Thirty-eight bladder tumors and corresponding urine sediment specimens were initially screened for 60 microsatellite markers from 18 chromosomes. Nine markers, most of which were different from those that had been used for Western patients, with frequent alterations in the initial patients were selected for further analysis. In the subsequent blinded experiment, microsatellite alterations were observed in urine sediments from 22 (96%) of 23 patients with bladder cancer and from all three patients with inverted papilloma. None of the urine sediments from the one patient with bladder lipoma, from the one patient with neurofibroma, or from the 12 individuals without evidence of bladder tumor showed any microsatellite alterations. CONCLUSIONS: Microsatellite analysis of urine sediments could be a practical method for detecting bladder cancer in the Chinese. Our identification of different microsatellite markers highlights possible ethnic and etiologic disparities between the Chinese and Western bladder cancer patients.     

Evaluation of microsatellite analysis in urine sediment for diagnosis of bladder cancer

Alterations at microsatellite DNA markers in cells exfoliated in urine have been correlated to the presence of bladder cancer. To check the feasibility of such noninvasive analysis to routinely diagnose bladder cancers, we have developed a highly sensitive method using fluorescent PCR to search for DNA microsatellite alterations in urine sediment compared with a blood paired sample. One hundred eighty-three patients were included in our study. This population comprised 103 bladder cancers (64 pTa stages), the complement representing controls and other benign or malignant diseases. Results of the analysis at 17 loci in a blinded study were compared with cystoscopy and/or pathology. The high reproducibility of this technique and the analysis of 26 control patients allowed us to determine for each microsatellite a cutoff characterizing a significant allelic imbalance. For bladder cancer detection, the overall sensitivity of the test was 84%. Using this procedure, we identified alterations in 81%, 84%, 91%, and 100% of pTa, pT1, pT2, and >pT2 stages, respectively. This corresponds to 79%, 82%, and 96% sensitivity for grades I, II, and III, respectively. Interestingly, for routine purposes, we observed an overall sensitivity of 80% (76% for pTa stages) when only the eight most rearranged microsatellites were considered. In conclusion, the noninvasive feature combined with the rapidity of this fluorescent and highly sensitive technique for the detection of early stages provides us with a useful help for the diagnosis of bladder cancer.     

Microsatellite instability in transitional cell carcinoma of the urinary tract and its relationship to clinicopathological variables and smoking

To determine whether microsatellite instability is involved in the development of transitional cell carcinoma (TCC) of the urinary tract, a microsatellite instability assay was carried out using PCR with 9 microsatellite loci. Thirty-eight TCC samples (30 patients with bladder cancer, 5 with renal pelvic tumors and 3 with ureteral tumors) and 1 lymph node with metastasis were examined. Microsatellite instability was found in 8 of 38 tumors examined, and 3 showed alterations in more than 2 microsatellite loci. All 8 tumors were beyond grade 2 and stage pT2 advanced tumors. Stages pT1-2 and pT3-4 patients differed significantly. Microsatellite instability was greater in smokers than non-smokers, but the differences were not significant. Microsatellite instability in TCC of the urinary tract is rare in superficial tumors but more common in invasive tumors. Microsatellite alterations would thus appear to occur, and possibly be importantly involved, in the tumorigenesis of urinary tract TCC. © 1996 Wiley-Liss, Inc.     

Genomic instability analysis of urine sediment versus tumor tissue in transitional cell carcinoma of the urinary bladder

Microsatellite alterations are a common feature of neoplastic cells. Our study aimed to compare the profile of microsatellite DNA alterations in tumor tissue and urine sediment at 12 selected microsatellite loci in transitional cell carcinoma of the bladder, and to determine which of the 12 markers or combination of markers has potential for the non-invasive diagnosis of bladder cancer. DNA alterations were examined using microsatellite markers on chromosomes 2p, 3p, 8p, 9p, 9q, 12q, 13q, 17p and 18q in 38 patients, including 12 with superficial Ta/T1 and 26 with muscle invasive T2-T4 bladder tumors. Microsatellite instability was a rare event in comparison with loss of heterozygosity and was related to a low rate of defects in mismatch repair genes. The sensitivity of microsatellite analysis was 75% (9/12) for Ta/T1 tumors and 69% (18/26) for T2-T4 tumors. Two tetranucleotide markers, D9S242 and D9S252, when combined, displayed microsatellite alterations in 59% (16/27) of microsatellite analysis-positive patients. DNA alterations were not detected in 21 non-tumor specimens. Twenty of 51 (39%) tumor DNA alterations were re-detected in urine sediments, and 7 alterations found in urine sediments were not found in the corresponding tumor specimens. No association was found between the DNA alterations and any of the prognostic parameters. However, the overall survival correlated with microsatellite alterations (P=0.04, log-rank test). These data suggest that markers at tetranucleotide repeats on chromosome 9q have particular diagnostic potential in bladder cancer. Moreover, microsatellite analysis is suitable for the selection of patients with a less favorable outcome.     

膀胱移行细胞癌中FHIT基因杂合性缺失及其蛋白表达的研究

目的分析候选抑癌基因FHIT及其蛋白表达在中国人膀胱移行细胞癌(transitional cell carcinoma,TCC)中的异常改变,探讨其潜在的临床意义.方法选取两个位于FHIT基因第5内含子的微卫星多态标记,对40例膀胱TCC患者的肿瘤组织和尿沉渣DNA进行杂合性缺失(loss of heterozygosity,LOH)分析,同时对相应的组织病理切片进行FHIT蛋白的免疫组化染色.结果在40例膀胱TCC患者的肿瘤组织中,两个微卫星多态标记D3S1234和D3S1300中至少有一个发生LOH的频率为57.8%;而在相应的尿沉渣DNA中,这两个位点中至少有一个发生LOH的频率为55.8%.同时在62.5%(25/40)的膀胱TCC肿瘤组织检出FHIT蛋白表达缺失;而在FHIT蛋白表达缺失的25例肿瘤组织中,有20例存在一个或两个上述微卫星位点的LOH.结论 FHIT基因在膀胱TCC患者的肿瘤组织和尿沉渣DNA中存在高频率LOH,提示这一基因在膀胱TCC的发生发展中可能起重要作用;杂合性缺失可能是膀胱TCC中FHIT蛋白表达下调的重要机理之一.而尿沉渣DNA中FHIT基因的LOH则有可能成为膀胱TCC早期诊断的潜在分子标志之一.     

检测尿脱落细胞中微卫星不稳定性的临床意义

目的:探讨膀胱癌患者尿脱落细胞微卫星不稳定性及临床意义.方法:将42例患者尿液标本经适当处理后,采用PCR-PAGE-银染方法检测尿脱落细胞中微卫星不稳定性的情况,分析其与临床病理参数的关系.结果:膀胱癌患者尿中MSI阳性率为59.52%(25/42),浸润性肿瘤患者尿中阳性率较高,但无统计学意义(P＞0.05);复发病例中阳性率(80.00%)比初发者(40.91%)明显要高(P＜0.05).结论:检测尿脱落细胞微卫星不稳定性可以作为膀胱癌的一种无创诊断及筛查手段.     

RT-PCR法检测尿脱落细胞中XIAP的表达在膀胱癌诊断中的价值

目的：比较膀胱癌患者尿液脱落细胞中XIAP表达的RT-PCR检测法和常规尿脱落细胞病理学检测在膀胱癌诊断中的临床价值。方法：采用逆转录聚合酶链反应技术（RT-PCR）检测51例膀胱尿路上皮癌患者尿液脱落细胞中XIAP-mRNA的表达,同时行常规尿脱落细胞病理学检测,20例非肿瘤人员作为对照组。结果：实验组51例尿脱落细胞XIAP-mRNA RT-PCR检测阳性27例（53%）,尿脱落细胞学病理学检测阳性12例（24%）,对照组20例尿脱落细胞XIAP-mRNA检测阳性1例（5.0%）,对照组尿脱落细胞病理学检测阳性0例（0%）。实验组RT-PCR检测膀胱尿路上皮癌患者尿脱落细胞中XIAP表达的敏感性高于尿脱落细胞病理学检测,差异有极显著统计学意义（P〈0.01）,实验组RT-PCR检测膀胱尿路上皮癌患者尿中XIAP表达的敏感性显著高于非肿瘤对照组,差异有极显著统计学意义（P〈0.01）。结论：膀胱尿路上皮癌患者尿脱落细胞中XIAP表达的RT-PCR检测法较常规尿脱落细胞病理学检测更敏感,临床上作为膀胱癌的筛选方法,有一定的临床价值。     

尿脱落细胞微卫星DNA改变在膀胱癌早期诊断中的应用

目的 利用检测尿脱落细胞微卫星ＤＮＡ序列（ｍｉｃｒｏｓａｔｅｌｌｉｔｅ,ＭＳ）改变,建立早期诊断膀胱癌的方法。方法 选择１０对微卫星ＭＳ引物,利用聚合酶链反应（ＰＣＲ）方法,以自身外周血和膀胱癌组织为对照,检测２８例膀胱癌患者尿脱落细胞中ＭＳ的失杂合（ｌｏｓｓ ｏｆ ｈｅｔｅｒｏｚｙｇｏｓｉｔｙ,ＬＯＨ）和不稳定性（ｍｉｃｒｏｓａｔｅｌｌｉｔｅ ｉｎｓｔａｂｉｌｉｔｙ,ＭＩＮ）。结果 ２８例膀胱癌患者中,２４例（８５．７％）尿脱落细胞至少在１个ＭＳ位点存在ＬＯＨ或ＭＩＮ改变,３例（１０．７％）脱落细胞学检查阳性患者尿脱落细胞均检出ＬＯＨ或ＭＩＮ。同一患者尿脱落细胞与癌组织ＬＯＨ改变一致率为９４．１％。１５例正常人悄脱落细胞中未见ＭＳ的改变。结论 利用检测尿脱落细胞ＭＳ改变诊断膀胱癌,比常规的细胞学检查更敏感、更     

尿液检查在膀胱癌诊断和术后复发监测中的现状及进展

膀胱癌是我国男性泌尿生殖系统肿瘤中的最常见肿瘤.膀胱镜检查是诊断和治疗后随访的最主要手段,但其往往会带给患者痛苦和恐惧.无创性的尿液检查替代膀胱镜诊断膀胱癌、监测复发及判断预后,一直是研究的热点.除尿细胞学外,目前通过尿液检查诊断和监测膀胱癌复发主要有尿核基质蛋白22、膀胱肿瘤抗原、免疫-细胞检查法、纤维素和纤维蛋白降解产物及荧光原位杂交技术等方法,但敏感性和特异性均不十分理想.检测尿液的端粒酶、透明质酸和透明质酸酶的方法其敏感性和特异性较好,但尚未经多中心的研究证实.对膀胱癌尿液中新的肿瘤标志物的初步研究结果表明,微卫星体异常、一些基因启动子的异常甲基化、细胞角蛋白、survivin等在膀胱癌的诊断中有较大的应用价值,但目前这些方法均有其局限性.合理地联合应用上述无创性尿液分析手段,可望推迟或减少膀胱镜检查,但尚难以取代膀胱镜.     

Clusterin as a Diagnostic and Prognostic Marker for Transitional Cell Carcinoma of the Bladder

We investigated the feasibility of profiling and measuring the concentration of clusterin in urine and serum for individuals with transitional cell carcinoma (TCC) of the bladder and comparing it with nontumor controls. In addition, we analyzed the correlation of expression of clusterin in specimens of TCC to various clinicopathologic parameters and prognosis of bladder cancer. Blood and urine samples were used from 68 patients with TCC of the bladder and from 61 patients with benign urological diseases. Enzyme-linked immunosorbent assays (ELISA) were performed for clusterin from serum and urine. Quantitation of clusterin mRNA was carried out in 68 bladder tumor specimens from radical cystectomy or transurethral resection and 26 normal bladder specimens from BPH patients by using RT-PCR method. Correlation for the expression of clusterin mRNA with clinicopathologic parameters was analyzed. Serum and urine clusterin was significantly higher in individuals with bladder cancer than control (p = 0.001). Sensitivity and specificity of serum and urine clusterin as a tumor marker for TCC of the bladder was found to be 80%, 91%, 87.1% and 96.7% respectively. Clusterin expression was significantly higher in TCC specimens than normal tissue specimens (P < 0.001). Expression of clusterin was significantly higher in patients with invasive TCC of the bladder than that in patients with superficial TCC and control (P < 0.001). Overexpression of clusterin mRNA was significantly associated with tumor recurrence and overall survival (p < 0.001). The recurrence-free survival time of patients with overexpression of clusterin was significantly shorter than that of patients with weak expression of clusterin (9.8 months vs. 35.2 months). Clusterin may be considered as a potential diagnostic and prognostic biomarker for bladder cancer using urine, serum and/or molecular biology techniques.     

Fetal fibronectin: a new screening-marker for bladder cancer?

Early detection of transitional cell carcinoma (TCC) of the urinary bladder is essential for effective treatment. While several serum markers have been evaluated, none have been widely accepted for practical clinical use. Thus, urinary markers have been introduced and investigated to detect the evidence of bladder cancer. But sensitivity and specificity range around 80% respectively. In a prospective study we evaluated fetal fibronectin in the urine of patients with TCC of the urinary bladder. The positivity of oncofetal fibronectin was measured in morning urine samples by membrane immunoassay. This FFN membrane immunoassay is a qualitative test, a solid-phase immunogold assay. A positive sample will result in a single spot after binding of the oncofetal fibronectin-immunogold complex to the membrane containing a monoclonal antibody specific to oncofetal fibronectin (FDC-6, which specifically recognizes III-CS region). The morning urine samples were collected from patients with TCC before they underwent transurethral resection (n=40, 34 non-invasive and 6 invasive carcinomas) and healthy controls (n=20). Oncofetal fibronectin was investigated in the surgical samples by immunohistochemistry (antibody FDC-6, APAAP technique). We found a positive result for oncofetal fibronectin in 38/40 patients with transitional cell carcinoma of the urinary bladder. Two patients with a small pTaG1-TCC showed negative results. In the urine of healthy controls no positive results were detected. Thus, there is a sensitivity of 95% and a specificity of 100%. The TCC was demonstrated as a source of oncfn. To our knowledge this is the first study showing that patients with an evident TCC have a demonstrable amount of oncofetal fibronectin in the urine. We conclude that a positive result is common in TCC-patients. The sensitivity and specificity of this test seems to be extraordinarily high. Because of the small number of cases further studies are required.     

RT-PCR法检测尿脱落细胞中XIAP的表达在膀胱癌诊断中的价值

目的:比较膀胱癌患者尿液脱落细胞中XIAP表达的RT-PCR检测法和常规尿脱落细胞病理学检测在膀胱癌诊断中的临床价值。方法:采用逆转录聚合酶链反应技术(RT-PCR)检测51例膀胱尿路上皮癌患者尿液脱落细胞中XIAP-mRNA的表达,同时行常规尿脱落细胞病理学检测,20例非肿瘤人员作为对照组。结果:实验组51例尿脱落细胞XIAP-mRNA RT-PCR检测阳性27例(53%),尿脱落细胞学病理学检测阳性12例(24%),对照组20例尿脱落细胞XIAP-mRNA检测阳性1例(5.0%),对照组尿脱落细胞病理学检测阳性0例(0%)。实验组RT-PCR检测膀胱尿路上皮癌患者尿脱落细胞中XIAP表达的敏感性高于尿脱落细胞病理学检测,差异有极显著统计学意义(P<0.01),实验组RT-PCR检测膀胱尿路上皮癌患者尿中XIAP表达的敏感性显著高于非肿瘤对照组,差异有极显著统计学意义(P<0.01)。结论:膀胱尿路上皮癌患者尿脱落细胞中XIAP表达的RT-PCR检测法较常规尿脱落细胞病理学检测更敏感,临床上作为膀胱癌的筛选方法,有一定的临床价值。