膀胱移行细胞癌尿脱落细胞CD44v6 mRNA表达的临床意义

目的 探讨膀胱移行细胞癌患者尿脱落细胞CD44v6mRNA的表达及其临床意义。方法 采用逆转录聚合酶链反应（RT—PCR）检测膀胱移行细胞癌37例,所有患者行经尿道膀胱肿瘤电切术或膀胱部分切除术;20例泌尿科非肿瘤患者以及10例健康志愿者尿脱落细胞CD44v6mRNA的表达设为对照组。结果 术前37例膀胱移行细胞癌中有29例尿脱落细胞CD44v6mRNA表达阳性,阳性率为78．4％,术后2周37例膀胱移行细胞癌尿脱落细胞CD44v6mRNA表达均为阴性;20例泌尿科非肿瘤患者以及10例健康志愿者尿脱落细胞CD44v6mRNA表达均为阴性。结论 尿脱落细胞CD44v6mRNA的表达阳性可以作为膀胱移行细胞癌的证据之一,有助于膀胱移行细胞癌的筛选和术后随访。     

尿路移行细胞癌微转移的检测

目的 探讨尿路移行细胞癌患者外周血CK20的表达及临床意义.方法 采用逆转录聚合酶链反应(RT-PCR)检测47例尿路移行细胞癌患者和14例健康志愿者以及18例非肿瘤患者外周血CK20的表达.以GAPDH作为内参照.结果 14例健康志愿者和18例非肿瘤患者外周血CK20的表达均为阴性;47例尿路移行细胞癌患者外周血CK20阳性表达率为21.3%(10/47):T1为0(0/29),T2为20%(2/10),T3为100%(4/4),T4为100%(4/4).随访12个月,6例(T2～T3)外周血CK20阳性的尿路移行细胞癌有3例(T3G2膀胱癌1例,TaG2和T3G2输尿管癌各1例)发生远处转移.结论 检测外周血CK20表达,可以提示癌细胞的血行播散,对判断预后以及指导治疗具有一定临床价值.     

晨尿脱落细胞角蛋白CK-20基因表达的检测及其临床意义

目的：探讨尿脱落细胞角蛋白（CK－20）基因表达的临床意义。方法：采用逆转录－聚合酶链反应技术，检测73例膀胱移行上皮细胞癌（TCC）患者（研究组）、20例泌尿系统非恶性肿瘤疾病患者（对照组）和21例正常人（正常组）晨尿脱落细胞CK－20基因表达，同时行尿脱落细胞学检查。结果：研究组尿脱落细胞CK－20基因表达的检出率为86．3％（63／73），明显高于尿脱落细胞学检查对TCC的检出率[20.5%(15/73)];对照组检出率为20．0％（4／20）；正常组检出率为0（0／21）。G3级TCC患者尿脱落细胞CK－20基因表达的检出率为100．0％（20／20）明显高于G1、G2级患者的76．5％（26／34）和89．5％（17／19）；T2－T4期尿脱落细胞CK－20基因表达的检出率为93．1％（27／29），明显高于Tis-T1期的81．8％（36／44）。结论：尿脱落细胞CK－20基因表达的检测有望成为1种灵敏、特异的TCC标志物。     

细胞角蛋白20对膀胱癌早期诊断的前瞻性研究

目的：评估细胞角蛋白20（CK20）标志物作为膀胱癌早期诊断及临床监测的价值。方法：对62例患者的尿液进行尿脱落细胞学及CK20标志物免疫荧光检测的前瞻性研究。分析参数包括肿瘤数目、大小及WHO分级，术前或活检前尿脱落细胞学和CK20标志物。结果：病理活检证实15例移行细胞癌中，CK20为13例阳性，2例阴性；47例非膀胱癌中，CK20标志物2例假阳性。与尿脱落细胞学比较，CK20标志物对膀胱移行细胞癌诊断的特异性和阳性预报值更高，分别为96．0％比82．5％（U＝2．18，P＜0．05），86．7％比52．4％（U＝2．16，P＜0．05）；但两者在敏感性和阴性预报值间无显著性差异（U值分别为：0．91和1．02，P＞0．05）。CK20表达与肿瘤分级间无明显相关性。结论：CK20是诊断膀胱移行细胞癌的一种良好标志物，其特异性明显优于尿脱落细胞学。     

尿核基质蛋白22和细胞角蛋白18在膀胱移行细胞癌中的表达及其临床意义

目的 探讨尿核基质蛋白22(NMP22)和细胞角蛋白18(CK18)在膀胱移行细胞癌中的表达及其临床意义.方法 采用酶联免疫吸附法(ELISA)对293例膀胱移行细胞癌、400例非移行细胞肿瘤、105例泌尿系良性病患者进行尿NMP22和CK18蛋白水平的检测.结果 膀胱移行细胞癌患者术前NMP22和CK18表达中位值分别为17.3 U/ml和484.2 U/L,非移行细胞肿瘤患者分别为6.8 U/ml和156.0 U/L,良性疾病患者分别为2.3 U/ml和66.6 U/L,3组之间进行比较,差异有统计学意义(P<0.001).以NMP22 10 U/ml和CK18 120 U/L为界值,其对膀胱移行细胞癌诊断的敏感度分别为79.2%和78.2%,特异度分别为88.6%和82.9%,两者联合检测的敏感度为91.7%.对术后患者动态观察,治疗有效患者的NMP22和CK18表达水平较治疗前明显下降,而复发、转移的患者则上升,差异有统计学意义(P<0.01).NMP22和CK18对膀胱移行细胞癌的检测有显著的相关性(r=0.689 P<0.0001).NMP22和CK18表达水平在不同病理分级和不同分期的患者中比较,差异有统计学意义(均P<0.01).结论 尿NMP22和CK18检测可作为膀胱移行细胞癌术前诊断、病情监测重要指标,两者联合检测可进一步提高敏感度.     

膀胱癌抗原在膀胱移行细胞癌诊断中的价值

目的评价膀胱癌抗原(UBC)对膀胱移行细胞癌(BTCC)的诊断价值.方法采用ELISA法对89例BTCC及108例泌尿系非移行细胞癌(TCC)患者的尿UBC进行检测,并同时行尿脱落细胞学检查.结果 BTCC患者尿UBC均值为30.5 μg/L,与泌尿系非TCC患者8.2μg/L的均值比较,差别有显著性意义(P＜0.01).尿UBC(以8.4μg/L为最适临界值)和尿细胞学诊断BTCC的敏感性分别为75.3%和16.9%,特异性分别为77.8%和100%,两者差别均有显著性意义(P＜0.01).结论尿UBC检测对BTCC的诊断是一种较为敏感、特异且无创的方法,敏感性明显优于尿细胞学,但临床上不能完全替代尿细胞学检查.     

环氧化酶-2 在膀胱癌组织及尿脱落细胞中的表达和意义

 目的 探讨COX-2在膀胱癌组织中的表达，了解尿脱落细胞COX-2表达在膀胱癌早期诊断中的价值。方法 应用免疫组化技术检测48例膀胱移行细胞癌组织、免疫细胞化学技术检测40例膀胱移行细胞癌患者和30例非肿瘤患者尿脱落细胞COX-2的表达。结果 膀胱移行细胞癌组织COX-2阳性表达率为72．9％，对照组正常膀胱黏膜无表达。COX-2的表达与膀胱癌临床分期显著相关(P<0．05)，不同病理分级膀胱癌的表达差别无显著性意义。非肿瘤患者尿脱落细胞无COx-2表达，膀胱癌尿脱落细胞COX-2免疫细胞化学检测阳性率为67．5％，明显高于常规尿细胞学的37．5％(P<0．05)，尤其对于G1级和Ta～T1期的低级、早期肿瘤，尿脱落细胞COX-2免疫细胞化学检测与常规尿细胞学检查相比，具有显著性意义(P<0．05)。结论 COX-2在膀胱移行细胞癌的发生发展中起重要作用，与肿瘤的浸润、转移相关。尿脱落细胞COX-2表达检测特异性高，可作为早期诊断膀胱癌的一种标志物。     

人膀胱癌细胞角蛋白20 mRNA表达的研究

目的　评估细胞角蛋白 2 0mRNA(CK2 0mRNA)是否可以作为膀胱癌临床诊断的一种有用标志物。方法　对 84例肉眼血尿患者的尿液进行尿脱落细胞学及CK2 0mRNA标记物RT/PCR检测。分析参数包括肿瘤数目、大小及WHO分级 ,术前或活检前尿脱落细胞学和CK2 0mRNA标志物。结果　病理活检证实 2 2例移行细胞癌中 ,CK2 0mRNA 18例为阳性 ,4例阴性 ;6 2例非膀胱癌患者中 ,CK2 0mRNA标志物 2例假阳性。与尿脱落细胞学比较 ,CK2 0mRNA标志物对膀胱移行细胞癌诊断的特异性和阳性预报值更高 ,分别为 96 .8%比 77.4 % (U =3.2 1,P <0 .0 1) ,90 %比 5 1.7% (U =2 .81,P <0 .0 1) ;但两者在敏感性和阴性预报值间无显著性差异 (U值分别为 :1.0 4和 1.2 1,P >0 .0 5 )。CK2 0mRNA表达与肿瘤分级间无明显相关性。结论　通过RT/PCR方法检测CK2 0mRNA是诊断膀胱移行细胞癌的一种良好生物标志物 ,其特异性明显优于尿脱落细胞学。     

蛋白质组学法检测膀胱移行细胞癌患者尿液中特异性肿瘤标志物

 目的:检测膀胱移行细胞癌患者尿液中差异表达蛋白质,筛选新的肿瘤标志物,进而探讨其与TCC发病机制的相关性,以及蛋白质组学法在膀胱移行细胞癌早期无创诊断方面的研究应用价值。 方法:选取青岛大学医学院附属医院泌尿外科TCC住院病人尿液标本24例,另外选取12例TCC术后病人、12例健康志愿者的尿液标本作为对照。采用蛋白质芯片技术结合表面增强激光解析/离子化-飞行时间-质谱技术检测各组尿液标本的差异表达蛋白质,然后到蛋白数据库中鉴定筛选肿瘤标志物。 结果:TCC组与对照组尿液标本的蛋白质存在差异表达,IMAC-Cu-3蛋白质芯片共发现6个蛋白及2个蛋白簇表达水平发生变化。联合检测3438Da及8002Da蛋白可使诊断TCC的敏感度达83.3%,特异性达75.0%。3438Da蛋白已被鉴定为α-defensin蛋白;搜索SWISS-PRO蛋白数据库,4310Da~5150Da蛋白簇为gp40蛋白。 结论:SELDI-TOF-MS蛋白质芯片技术是一种快速、简便易行、用量少和高通量分析方法,能直接筛选出膀胱移行细胞癌患者尿液中特异的肿瘤标志物,为蛋白组学方法在膀胱移行细胞癌早期无创诊断、判断预后及探讨发病机制方面的研究应用开拓了广阔前景。     

膀胱移行细胞癌患者尿脱落细胞中MUC7的表达及其临床意义的研究

目的:探讨膀胱移行细胞癌(BTCC)患者尿脱落细胞中MUC7的表达及其临床意义.方法:采用逆转录聚合酶链反应(RT-PCR)检测42例BTCC患者、40例对照者尿脱落细胞中MUC7的mRNA表达,β-actin作为内参照.结果:40例对照组中有2例MUC7 mRNA表达阳性,阳性率为5.0%;42例BTCC患者中36例尿脱落细胞中MUC7 mRNA表达阳性,阳性率为85.7%,其诊断特异性为95%.不同临床分期尿脱落细胞中MUC7 mRNA的阳性表达率:Ta-T1期为64.3%(9/14),T2-4期为96.4%(27/28).浸润性BTCCMUC7的阳性表达率显著高于浅表性BTCC(P＜0.05).不同病理分级BTCC患者尿脱落细胞中MUC7的mRNA的阳性表达率为G158.3%(7/12),G2 93.8%(15/16),G3 100%(14/14).MUC7mRNA的表达与BTCC的病理分级呈正相关.MUC7诊断BTCC的阳性预测值为94.7%,阴性预测值为86.3%.结论:BTCC患者尿脱落细胞中MUC7的阳性表达率明显高于对照组.尿脱落细胞中MUC7表达与临床分期、病理分级呈正相关.采用RT-PCR法检测尿脱落细胞中MUC7表达诊断BTCC的敏感性和特异性好、阳性预测值高,特别对T2-4期肿瘤的诊断敏感性好.     

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。     

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.     

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。     

Telomerase activity in patients with transitional cell carcinoma: a preliminary study

BACKGROUND: Telomerase activity is not detectable in normal cells, and their telomers shorten until the chromosome is unable to replicate. Immortal cells have short but stable chromosomes and increased telomerase activity. Transitional cell carcinoma (TCC) has only a few useful markers of diagnostic or prognostic importance. The objective of this study was to determine whether there was a correlation between telomerase activity and the grade or stage of TCC, and whether the enzyme's activity could serve as a biochemical marker of this tumor. METHODS: The study included 29 patients with TCC. From each patient, samples of urine cells were obtained, and a cup biopsy was taken from an apparently normal area as well as from a part of the bladder tumor resected transurethrally. Control uroepithelial biopsies were taken from normal transitional cell sites from non-TCC patients. Biopsies or cells were subjected to either histologic examination or telomerase activity determination. RESULTS: Twenty-six of 29 (90%) of the tumor biopsies exhibited telomerase activity. Most of the cup biopsies were categorized as metaplastic or dysplastic, and 20 of 29 (69%) of these exhibited telomerase activity. Telomerase activity was found in 17 of 21 (81%) of the urine cells but in only 3 of 14 (21%) of control urine cells. All (10 of 10) of the uroepithelial biopsies taken from non-TCC patients did not show any telomerase activity. CONCLUSIONS: In this study, almost all tumor biopsies exhibited telomerase activity. The high incidence of telomerase activity found in cup biopsies of the malignant field uroepithelial cells from cup biopsies of TCC patients may suggest that telomerase could be activated early in carcinogenesis. A high incidence of telomerase activity was found in voided uroepithelial cells of TCC patients; however, no correlation between this activity and the histologic determination of grading and staging of the tumor was found.     

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.     

CK20m RNA在大肠癌病人外周血中的表达及其临床意义

目的：以CK20mRNA作为靶产物,检测大肠癌病人外周血中的癌细胞即微转移。并探讨其存在的临床意义。方法：应用逆转录聚合酶链反应（RT-PCR）。检测20例健康志愿者和47例大肠癌患者外周血中CK20mRNA的表达。同时检测20例大肠癌新鲜组织标本中CK20mRNA的表达。结果：20例大肠癌组织标本均扩增出370bp的条带。结肠腺癌HT-29细胞系作为阳性对照亦扩增出370bp的条带,20例正常人外周血中CK20mRNA表达均阴性。全组47例大肠癌病人,19例外周血中表达CK20mRNA,总阳性率为40.4%(19/47),CK20mRNA的表达与肿瘤Dukes分期,最大直径,肝转移有关,结论：CK20mRNA可作为检测大肠癌病人外周血中微转移的良好指标;外周血中CK20mRNA的检测有助于识别出具有复发或远处转移倾向的高危大肠癌病人。