粘蛋白7在膀胱癌尿脱落细胞中的表达及意义

目的:探讨粘蛋白7(mucin7,MUC7)在膀胱移行细胞癌(TCC)诊断中的意义。方法:采用巢式逆转录聚合酶链反应(nested RT-PCR)方法检测MUC7mRNA在TCC患者尿脱落细胞中的表达。结果:68例TCC患者中有42例尿液MUC7表达阳性,敏感性为62%;MUC7表达与临床分期有关而与病理分级无关。对照组25例中有3例在尿液中MUC7为阳性表达,特异性为88%。结论:用RT-PCR的方法检测尿脱落细胞中的MUC7,具有较高的膀胱癌诊断价值。     

联合检测尿液中Survivin和MUC7在膀胱癌诊断中的临床应用

目的探讨检测尿液中Survivin和mucin 7（MUC7）在诊断膀胱癌中的临床价值。 方法对75例膀胱肿瘤患者、38例泌尿系统良性疾病患者及20例健康自愿者在行膀胱镜检前分别留置3份尿液标本,通过检测尿液中Survivin、MUC7和尿脱落细胞学的敏感性和特异性,分析比较3种方法在临床诊断膀胱癌的临床应用价值。 结果Survivin诊断膀胱癌的敏感性和特异性分别是93.3%和98.3%,MUC7诊断膀胱癌的敏感性和特异性分别是80%和100%;尿脱落细胞学诊断膀胱癌的敏感性和特异性分别是66.7%和100%;Survivin联合MUC7的敏感性和特异性分别是98.7%和100%;Survivin联合MUC7、Survivin、MUC7与尿脱落细胞学的敏感性比较差异均有统计学意义（P<0.05）。Survivin联合MUC7、Survivin、MUC7与尿脱落细胞细胞学的准确性分别是99.2%、95.5%、88.7%、81.2.3%,与脱落细胞学组比较,差异有统计学意义（P<0.05）。 结论联合检测两种肿瘤标记物Survivin及MUC7可以提高膀胱癌诊断的敏感性和准确性。     

CK-20在膀胱癌术后随访中的临床意义

目的　评估尿脱落细胞CK -20表达的检测在诊断膀胱癌复发中的价值及其临床意义。方法　对 60 例膀胱癌术后随访尿液标本进行CK -20标志物检测。分析参数包括肿瘤数目、大小、WHO分级及 CK- 20 mRNA标志物。结果　20 例复发者中 12例CK -20表达阳性(60%),且均为血尿标本; CK 20 表达检测诊断膀胱癌复发的敏感性、特异性及准确性分别为 60%、90%及80%; CK -20的表达与肿瘤分级间无明显相关性。结论　CK- 20 检测可以作为监测膀胱癌术后随访病人肿瘤复发的良好标记物。     

Qualitative and quantitative detection of urinary human complement factor H-related protein (BTA stat and BTA TRAK) and fragments of cytokeratins 8, 18 (UBC rapid and UBC IRMA) as markers for transitional cell carcinoma of the bladder

OBJECTIVE: To evaluate the role of BTA stat, BTA TRAK, UBC Rapid, UBC IRMA and voided urinary cytology in the detection of bladder transitional cell carcinoma (TCC). METHODS: The study included 78 patients with TCC of the bladder (group A), 62 patients with a history of bladder TCC without tumor recurrence at the time of examination (B, control group), 20 patients with other malignancy of the urinary tract (C), 38 patients with non-malignant urinary tract diseases (D), 10 patients with urinary tract infection (E) and 10 healthy volunteers (F). Except in group F, voided urine was collected before cystoscopy or cystectomy. RESULTS: The specificity and sensitivity in bladder cancer detection were 87.1 and 74.4%, respectively with BTA stat, 79.3 and 48.7%, respectively with UBC Rapid, 100 and 33.3%, respectively with cytology, 72.6 and 75.6%, respectively with BTA TRAK, 64.5 and 70.5%, respectively with UBC IRMA. CONCLUSIONS: The BTA stat and BTATRAK tests are superior to UBC Rapid, UBC IRMA and urinary cytology in detection of bladder TCC. In daily practice however cytology remains the best adjunct to cystoscopy because of its high sensitivity in Tis and 100% specificity. Cystoscopy cannot be replaced by any of evaluated methods.     

Clinical utility of a multiprobe FISH assay in voided urine specimens for the detection of bladder cancer and its recurrences,compared with urinary cytology

OBJECTIVES: To evaluate the clinical utility of a Multi-color FISH (fluorescence in situ hybridization) assay in voided urine specimens for the detection of bladder cancer and its recurrences, comparing the results with those afforded by urinary cytology. METHODS: Voided urine samples from 86 patients were obtained for urine cytology and FISH analysis. The latter was performed using a mixture of fluorescent labeled DNA probes for the centromeric regions of chromosomes 3, 7 and 17, and the 9p21 region. Cystoscopy with biopsy or tumor resection was performed in all patients, comparing the pathological results with the cytological and FISH findings. RESULTS: Urinary cytology affords an overall sensitivity of 63.8%, the figure being 25% for grade 1, 66.6% for grade 2 and 94.7% for grade 3 tumors. The sensitivities for FISH were 53.3% for grade 1, 83.3% for grade 2 and 100% for grade 3 tumors, with an overall sensitivity of 80.4%. The specificities of urinary cytology and FISH were 86.1 and 85.3%, respectively. CONCLUSIONS: FISH improves the sensitivity rates obtained with urine cytology for bladder cancer detection in all tumor grades and stages, and offers similar specificity. FISH doubles the accuracy of urinary cytology in application to low grade-stage tumors, and detects all high grade infiltrating tumors.     

核基质蛋白22检测与尿脱落细胞学检查在膀胱癌诊断中的价值

目的　探讨尿核基质蛋白 2 2 (NMP 2 2 )检测和尿脱落细胞学检查在膀胱移行细胞癌诊断中的价值。　方法　对 15 5例怀疑膀胱癌者进行尿NMP 2 2与尿细胞学检查 ,其中 95例经组织学证实为膀胱移行细胞癌。比较两者诊断膀胱癌的敏感性和特异性。　结果　尿NMP 2 2的敏感性为6 5 .3%、特异性为 70 .0 % ;尿细胞学的敏感性为 4 3.2 %、特异性为 83.3%。NMP 2 2在膀胱癌不同分级和分期中的敏感性优于尿细胞学 (P <0 .0 5 )。　结论　尿NMP 2 2检测在早期诊断膀胱癌方面优于尿细胞学检查 ,可以作为膀胱癌的早期检测指标。     

单抗免疫分析法在血尿及膀胱移行细胞癌诊断中的应用

目的 研究单抗免疫分析法（ＢＴＡ Ｓｔａｔ）在血尿及膀胱肿瘤诊断中的准确性和可靠性。方法 采用ＢＴＡ Ｓｔａｔ检测试剂盒对６５例无痛血尿患者,７１例膀胱移行细胞癌电切术后患者及１０例正常人进行了ＢＴＡ Ｓｔａｔ检测。     

尿脱落细胞CK20检测在膀胱癌诊断中的价值

目的:检测膀胱癌患者尿脱落细胞细胞角蛋白20(CK20)表达情况并探讨其在膀胱癌诊断中的价值。方法:应用RT-PCR方法分别检测35例膀胱癌患者、8例泌尿生殖系良性疾病患者及4例健康自愿者尿脱落细胞CK20表达情况,并与尿脱落细胞学检查结果进行比较。结果:35例膀胱癌患者中有26例的自排尿标本中可检测出CK20(敏感度为74.29％)。尿脱落细胞学检测阳性例数则为12例(敏感度为34.29％)。前者与后者比较差异有显著性意义(P<0.05)。8例泌尿生殖系良性疾病患者及4例健康自愿者的自排尿标本中无一例可检测出CK20(特异性为100％)。不同分化程度及不同临床病理分期膀胱癌患者之间尿脱落细胞(2K20阳性表达率的差异有显著性意义(P<0.05)。结论:尿脱落细胞CK20检测的敏感性及特异性均高,有望成为膀胱癌诊断和术后复发监测的一种有效手段。     

Comparison of cytology and nuclear matrix protein 22 for the detection and follow-up of bladder cancer

OBJECTIVES: This study was designed to determine the clinical usefulness of the nuclear matrix METHODS: 84 patients suffering from bladder cancer or suspected bladder cancer, 25 patients with benign urological lesions and 60 healthy controls participated in a prospective study. Freshly voided spot urine samples were taken for cytological examination and determination of NMP 22 levels by enzyme-linked immunoassay. RESULTS: The sensitivity of the NMP 22 test according to the tumor grading was (results of cytology in brackets): G1 25.0% (20.0%); G2 68.2% (59.1%), and G3 100.0% (66.7); overall sensitivity was 62.5% (45.0%). The sensitivity for superficial bladder cancer was 46.7% (36.7%) and for invasive bladder cancer 90.0% (70.0%). The specificity was 65.9% (88.9%). CONCLUSIONS: NMP 22 is a reliable tool for detecting invasive bladder cancer. Results for the frequently occurring low grade superficial bladder cancer are as poor as those obtained with cytology. In addition benign lesions such as urolithiasis or urinary tract infection lead to false-positive results. Therefore cystoscopy has to be performed when trying to detect and follow-up bladder cancer.     

荧光原位杂交技术检测膀胱尿路上皮癌尿液的临床应用研究

目的：评价荧光原位杂交技术（fluorescence in situ hybridization,FISH）检测膀胱尿路上皮癌患者尿液的应用价值。方法：收集我院2007年10月-2009年4月期间77例膀胱尿路上皮癌患者、43例非尿路上皮癌的血尿患者（通过膀胱镜检查排除尿路上皮癌）和泌尿系良性疾病患者的晨尿,同时行FISH检测和尿脱落细胞学分析,再结合病理结果将两种方法进行比较。FISH检测使用荧光标记DNA探针混合物与细胞核上3、7、17号染色体着丝粒和9p16位点进行杂交。结果：FISH总的敏感度和特异度分别为89．6％和95．3％,G1-3各级的敏感度分别为76．1％、90．9％、100％,Ta、Tis、T1、T2-4各期的敏感度分别为55．6％、100％、88．9％、97．4％。尿脱落细胞学分析总的敏感度和特异度分别为37．7％和93．0％,G1-3各级的敏感度分别为0％、33．3％、78．3％,Ta、Tis、T1、T2-4各期的敏感度分别为11．1％、100％、14．8％、56．4％。结论：FISH比尿脱落细胞学提高了膀胱癌患者检测的敏感度,而特异度两者相近。FISH使低级别浅表型膀胱癌的准确率明显提高,几乎能检测出所有高级别的浸润型膀胱癌。相对于尿脱落细胞学,FISH检测更佳。     

IMMUNOSTAINING OF LEWIS X IN CELLS FROM VOIDED URINE, CYTOPATHOLOGY AND ULTRASOUND FOR NONINVASIVE DETECTION OF BLADDER TUMORS

Purpose

We examined the use of immunostaining of the Lewis X antigen in exfoliated cells from voided urine samples, cytopathology and bladder ultrasound for noninvasive detection of bladder tumors as a potential substitute for cystoscopy.

Materials and Methods

A total of 260 patients were included, of whom 80 were evaluated because of irritative symptoms or hematuria and 180 were examined during followup visits after resection of bladder tumors. Voided urine samples were obtained from each patient for immunocytology and cytopathology. Bladder ultrasound and cystoscopy were performed. Biopsies were obtained whenever a bladder tumor was seen or if carcinoma in situ was suspected. Indirect immunoperoxidase staining was done on cytocentrifuge slides, using the P12 monoclonal antibody against the Lewis X antigen.

Results

Cystoscopy and biopsies revealed bladder tumors in 84 patients. Immunocytology of 1 urine sample resulted in a sensitivity of 79.8% and a specificity of 86.4%. The diagnosis of primary carcinoma in situ by immunocytology was correct in 100% of the cases. The examination of 2 consecutive urine samples detected 95.1% of the tumors. False-negative results occurred in a few cases with small, superficial, low grade tumors. Cytopathology and bladder ultrasound resulted in a sensitivity of 47.6 and 66.7%, and a specificity of 97.7 and 97.2%, respectively. The results of immunocytology of 2 urine samples were equivalent to the combination of immunocytology of a single urine sample, cytology and ultrasound.

Conclusions

Immunostaining of the Lewis X antigen is significantly more sensitive than cytopathology for the detection of low grade as well as high grade tumor cells in voided urine. Immunocytological evaluation of 2 consecutive voided urine specimens for the Lewis X antigen is the most sensitive method currently available for noninvasive detection of transitional cell tumors. This assay may replace cystoscopy for detection of bladder cancer.     

ProteinChip technology reveals distinctive protein expression profiles in the urine of bladder cancer patients

OBJECTIVE: Since accurate biomarkers for the early diagnosis or individual prognosis of the bladder carcinoma are still not available, we used the ProteinChip technology, to search for discriminating protein expressions associated with this cancer and its subtypes. METHODS: A training set consisting of 30 archival urine samples from bladder carcinoma patients and 30 urinary samples from healthy volunteers, was analyzed via ProteinChip technology and computer based data mining. Mass clusters of differentially expressed proteins were verified by a second set (test set) comprising 21 bladder carcinoma urine samples and 21 non-tumor urinary samples. Expression differences between carcinoma subtype sample groups of the initial training set were assessed by a trend test. RESULTS: Bladder carcinoma was segregated from control with a sensitivity and specificity of 80% and 90 to 97% in the trainings set, as well as 52 to 57% and 57 to 62% in the test set, respectively. Segregation of pooled tumor stages pT2-pT3 from stages pT1 and pTa was possible at the 53.3 kDa cluster of the CM10-chip array data derived rule base. CONCLUSION: ProteinChip technology together with adapted computer based data mining tools are useful for the rapid establishment of potential protein biomarkers.     

膀胱移行上皮癌患者尿及癌组织中Survivin表达的临床意义

目的：探讨Survivin对膀胱癌早期发现、常规筛选且无损伤的方法,研究其与肿瘤病理分级（期）的相关性。方法：留取53例膀胱移行上皮癌、20例泌尿系其他疾病、10例健康志愿者新鲜中段晨尿,同上53例术后癌组织。利用巢式RT—PCR、实时定量PCR技术检测Survivin的表达,同时行尿脱落细胞学及膀胱镜病检。结果：53例膀胱癌患者尿及癌组织中Survivin均有表达,敏感性为100％,特异性为90％。与尿脱落细胞学敏感性比较两者差异有统计学意义（P〈0．05）,与膀胱镜检比较差异无统计学意义。三种方法特异性比较差异无统计学意义（P〉0．05）。癌组织中Survivin的量与肿瘤病理分级（期）正相关（P〈0．01）。结论：检测尿脱落细胞中Survivin的表达有望成为临床诊断、筛检膀胱癌的较可靠方法。Survivin在膀胱的演进过程中可能起重要作用,可望作为检测膀胱癌恶化进展的指标。     

Comparative evaluation of the nuclear matrix protein,fibronectin, urinary bladder cancer antigen and voided urine cytology in the detection of bladder tumors

PURPOSE: We evaluate the diagnostic efficacy of nuclear matrix protein-22 (NMP22, Matritech, Newton, Massachusetts), fibronectin and urinary bladder cancer antigen (UBC, IDL Biotech, Borlange, Sweden) compared with voided urine cytology in the detection of bladder cancer. MATERIALS AND METHODS: A total of 168 patients provided a single voided urine sample for NMP22, fibronectin an ideal monoclonal for urinary bladder cancer and cytology before cystoscopy. Cystoscopy was done for all patients as the reference standard for identification of bladder cancer. Biopsy of any suspicious lesion was performed for histopathological examination. Of the 168 cases 100 were histologically diagnosed as bladder cancer, whereas the remaining 68 had benign urological disorders. A group of 47 healthy volunteers were also enrolled in this study. Voided urine was evaluated by NMP22, fibronectin and UBC, and their values were expressed relative to mg. creatinine. RESULTS: The optimal threshold values for NMP22, fibronectin and UBC were calculated by receiver operator characteristics curves as 27 units per mg. creatinine, 198 mg./mg. creatinine and 13 ng./mg. creatinine, respectively. The levels and positive rates of the 3 parameters were significantly higher in the malignant group compared to either the benign group or normal controls. Of the entire group NMP22, fibronectin and UBC were positive in 93.2%, 91% and 68.2%, respectively in bladder cancer cases with positive cytology. Moreover, these positive rates were significantly higher in bilharzial bladder cancer cases (58.8%, 67.5%, 58.8%, respectively) compared to nonbilharzial cases (35.6%, 36.3%, 31.1%). Overall sensitivity and specificity were 85% and 91.3% for NMP22, 83% and 82.6% for fibronectin, 67% and 80.8% for UBC and 44% and 100% for voided urine cytology. Combined sensitivity of voided urine cytology with the 3 biomarkers together was higher than either combined sensitivity of voided urine cytology with 1 of the biomarkers or than that of the biomarker alone. CONCLUSIONS: Our data indicate that NMP22 and fibronectin had superior sensitivities compared to UBC and voided urine cytology, while NMP22 and voided urine cytology had the highest specificities. The combined use of markers increased the sensitivity of cytology from 44% to 95.3%. The higher sensitivities of markers in bilharzial than nonbilharzial bladder cancer highlight their clinical use in screening patients with urinary bilharziasis.     

The simultaneous use of telomerase, cytokeratin 20 and CD4 for bladder cancer detection in urine

OBJECTIVE: 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. METHODS: 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. RESULTS: 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%). CONCLUSIONS: 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.     

Urinzytologie beim Harnblasenkarzinom

ImmunoCyt is a new immunocytologic fluorescence test promising a higher diagnostic sensitivity, esp. for TaG1 carcinomas. The aim of the study was to evaluate the sensitivity of the test in diagnosis of bladder cancer as compared to both urinary cytology and histopathology. A total of 121 spontaneous urine samples of 92 patients (age range 28 to 86, mean 62.5 years) was examined. 41 of the samples were of patients suspicious of transitional cell carcinoma, 46 of patients in whom symptoms were suggestive of tumor recurrence, and 34 of patients who were part of a follow-up protocol. Cystoscopy was performed in all patients. The ImmunoCyt-test was carried out according to the manufacturers protocol using 3 fluorescent monoclonal antibodies. A total of 113 specimens could be evaluated. In 87 cystoscopy and/or histology was negative (control group). There was histologic evidence of 7 pTaG1, 4 pTaG2, 8 pT1G2/G3, and 7 pT2G2/G3 bladder cancers. As for ImmunoCyt and cytology specificity was 83.9% and 91.9%, resp. The combined specificity was 81.6%. Sensitivity amounted to 38.5% and 34.6%, resp., the combined sensitivity to 53.8%. Sensitivity for TaG1 carcinomas was 14.3% each, for TaG2 carcinomas 25% and 50%, for T1G2/G3 carcinomas 37.5% each, and for T2G2/G3 carcinomas 71.4% and 42.9%, resp. In our study the ImmunoCyt test did not show the expected increase in the detection of TaG1 bladder cancers. Because of false-positive results the test should only be used in combination with urinary cytology which reveals a higher specificity. In conclusion the ImmunoCyt test can not replace cystoscopy (with biopsy) in diagnosis and monitoring of bladder cancer.     

Muskelinvasives Urothelkarzinom der Harnblase

Detection of metastases in lymph nodes is an important prognostic factor for progression-free survival in bladder cancer patients. Patients undergoing radical cystectomy with pelvic lymphadenectomy are randomized in the LEA study (AUO AB 25/02) into two groups receiving standard (obturator and external nodes) or extended lymphadenectomy (complete pelvic nodes up to the inferior mesenteric artery).The aim of this study is the detection of lymph node metastases that are not identified with classic pathological methods using RT-PCR as a highly sensitive and specific method. For detection of occult disseminated tumor cells we analyze the expression of the tumor markers cytokeratin 20 (CK-20), uroplakin II (UP II), mucin 2 (MUC2), and mucin 7 (MUC7).We examined 315 lymph nodes from 19 cystectomy patients for the expression of CK-20. In 93 lymph nodes CK-20 expression was detected whereas only 18 lymph nodes were histopathologically positive. More than one third of CK-20-positive lymph node metastases were located outside the standard lymphadenectomy field. We did not detect any skip lesions. Follow-up data will validate if there is a correlation between detection of occult disseminated tumor cells and progression-free survival.     

几种新瘤标对膀胱癌早期诊断价值的比较

目的：比较几种新瘤标对膀胱移行细胞癌早期诊断的价值。方法：对322例肉眼或镜检血尿、膀胱刺激症状或发现膀胱占位者，在作膀胱镜检查前行尿液中细胞角蛋白20(CK20)、核基质蛋白(NMP22)、ImmunoCyt、膀胱肿瘤抗原(BTA stat)检测和常规尿细胞学检查，比较它们的敏感性和特异性。结果：经膀胱镜和病理检查诊断膀胱移行细胞癌149例，其中浅表性肿瘤110例，浸润性39例；G181例，G246例．G322例。CK20、NMP22、ImmunoCyt、BTA stat、尿细胞学和膀胱镜检的敏感性分别为90．6％、85．2％、82．6％、65．8％、30．2％和94．6％，特异性分别为83．6％、84．0％、78．1％、64％、100％和100％。结论：新瘤标CK20、NMP22、ImmunoCyt具有较高的敏感性，与BTA stat和尿细胞学比较，差异有统计学意义，可用于膀胱肿瘤的早期诊断。