尿脱落细胞LewisX检测诊断膀胱尿路上皮癌的价值

目的：探讨LewisX抗原在膀胱尿路上皮癌非侵袭性诊断中的应用价值。方法：采用EnVision免疫细胞化学方法，检测52例膀胱尿路上皮癌和16例非肿瘤患者尿脱落细胞标本中LewisX抗原的表达情况，并与细胞病理学检测结果相比较。结果：尿路上皮癌诊断的敏感性和特异性分别为84．6％和87．5％，其敏感性显著高于细胞病理学。结论：尿脱落细胞LewisX抗原免疫染色，是检测膀胱尿路上皮癌可行的较敏感的非侵袭性方法。     

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。     

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。     

CD15在膀胱癌尿脱落细胞学检测中的应用价值

目的：探讨CD15在膀胱癌尿脱落细胞学诊断中的应用价值。方法：采用EnVision免疫细胞学染色方法，检测52例膀胱尿路上皮癌和16例非肿瘤病人尿脱落细胞标本中CD15的表达情况，并与细胞病理学检测相比较。结果：以5％尿脱落上皮细胞CD15染色阳性为膀胱癌诊断阈值，其敏感性和特异性分别为84．6％和87．5％，敏感性显著高于细胞学检查。结论：尿脱落细胞CD15免疫染色，有助于膀胱尿路上皮癌的诊断。     

荧光原位杂交在膀胱尿路上皮癌中的应用

目的探讨3,7,17号染色体及9p21(p16基因)组合探针在监测膀胱尿路上皮癌术后复发及诊断膀胱尿路上皮癌的应用价值。方法利用荧光原位杂交（FISH）检测膀胱尿路上皮癌患者的尿脱落细胞及组织切片中3,7,17号染色体及9p21组合的畸变情况。结果FISH监测膀胱尿路上皮癌术后复发的敏感度为87.50%;术前FISH阳性的患者术后更易复发;FISH诊断膀胱尿路上皮癌的阳性率为78.85％。结论利用FISH检测尿脱落细胞中3,7,17号染色体及9p21组合的畸变能有效辅助监测膀胱尿路上皮癌术后复发;并可能在一定程度上预测术后复发;同时利用FISH检测组织切片中3,7,17号染色体及9p21组合的畸变也能有效辅助膀胱尿路上皮癌的诊断。     

膀胱移行上皮癌尿脱落细胞端粒酶活性检测

[目的]检测尿脱落细胞端粒酶活性表达对膀胱移行上皮癌的诊断价值.[方法]应用PCR为基础的端粒重复片段扩增方法(TRAP-Hyb Kit)分别检测52例膀胱移行上皮癌患者及10例泌尿生殖系良性疾病患者尿脱落细胞端粒酶活性表达情况,并与尿脱落细胞学检查结果进行比较.[结果]52例膀胱移行上皮癌患者中有34例的自排尿标本中可检测出端粒酶活性表达(65.4%);尿脱落细胞学检查阳性例数为11例(11/30,36.7%);两者比较差异有显著性(P＜0.05).泌尿生殖系良性疾病的尿脱落细胞端粒酶的阳性率为10.0%.但端粒酶活性与肿瘤的分级无相关性.[结论]尿脱落细胞端粒酶活性检测作为无创性的检测方法,可作为膀胱移行上皮癌早期诊断、术后监测的一种有效手段.     

尿路肿瘤细胞多重荧光PCR检测微卫星不稳定

目的：研究膀胱肿瘤尿路脱落细胞的微卫星状态及其临床病理关系。方法：35例膀胱肿瘤尿液脱落细胞应用多重荧光PCR检测尿路的微卫星状态。结果：35例尿液样本：膀胱肿瘤组15例,有6例MSI-H,8例MSI-L,1例MSS,诊断阳性敏感度达93.33%;膀胱癌术后复查组6例,血尿及尿路上皮轻度异型组7例,正常体检组7例中,在血尿及轻度异型组发现1例MSI-L,其余均为MSS,阴性特异率达95%。结论：尿路脱落细胞的微卫星检测可作为膀胱肿瘤诊断和监测复发的有效非侵袭性检测方法。     

尿路肿瘤细胞多重荧光PCR检测微卫星不稳定

目的:研究膀胱肿瘤尿路脱落细胞的微卫星状态及其临床病理关系。方法:35例膀胱肿瘤尿液脱落细胞应用多重荧光PCR检测尿路的微卫星状态。结果:35例尿液样本:膀胱肿瘤组15例,有6例MSI-H,8例MSI-L,1例MSS,诊断阳性敏感度达93.33%;膀胱癌术后复查组6例,血尿及尿路上皮轻度异型组7例,正常体检组7例中,在血尿及轻度异型组发现1例MSI-L,其余均为MSS,阴性特异率达95%。结论:尿路脱落细胞的微卫星检测可作为膀胱肿瘤诊断和监测复发的有效非侵袭性检测方法。     

长链非编码RNA HOTAIR抑制膀胱尿路上皮癌对阿霉素的敏感性

目的:研究长链非编码RNA HOTAIR在膀胱尿路上皮癌中的表达,明确其对膀胱尿路上皮癌对阿霉素敏感性的调控作用.方法:HOTAIR基因在膀胱尿路上皮癌及癌旁组织中的表达由实时定量PCR法检测.将HOTAIR基因的表达载体和沉默载体分别转染膀胱尿路上皮癌J82细胞,实时定量PCR验证转染效果.应用MTT法检测HOTAIR基因表达改变对于J82细胞的增殖能力及对阿霉素敏感性的调控作用.结果:HO-TAIR基因在膀胱尿路上皮癌表达显著上调.HOTAIR基因的表达载体和沉默载体能够显著上调或沉默HO-TAIR基因的表达.HOTAIR高表达能够促进J82细胞增殖,抑制J82细胞对阿霉素的敏感性;而HOTAIR表达沉默的J82细胞增殖能力下调明显,对阿霉素的敏感性明显增加.结论:长链非编码RNA HOTAIR在膀胱尿路上皮癌中高表达,HOTAIR能够作为癌基因促进膀胱尿路上皮癌的细胞增殖能力并抑制膀胱尿路上皮癌对阿霉素敏感性.     

尿液核基质蛋白22在膀胱移行上皮癌筛检中的价值

目的：评价尿核基质蛋白22(NMP22)在膀胱移行上皮癌筛检中的意义。方法：采用酶联免疫法检测28例膀胱移行上皮癌、25例泌尿系良性疾病和10例泌尿系非移行上皮癌患者尿NMP22水平，并与尿脱落细胞学检查进行比较。结果：膀胱移行上皮癌患者尿NMP,,中位值为66．5u／ml，明显高于泌尿系良性疾病和非移行上皮癌，与肿瘤分期、分级、数目无关，以10u／ml为临界，诊断膀胱癌敏感性为85．7％，特异性为60％，尿脱落细胞学检查敏感性为32．1％，特异性为100％。结论：尿NMP22检测膀胱移行上皮癌敏感性、特异性均较高，可用于膀胱移行上皮癌的筛选和术后随访。     

Bladder cancer detection and surveillance: carcinoembryonic antigen as a monitor of neoplastic transformation

Routine cytopathology of exfoliated urothelial cells will identify only 70% of patients with transitional cell carcinoma (TCC) because most well-differentiated bladder tumors are not detected. Carcinoembryonic antigen (CEA) was identified on urothelial cancer cells from 66% of patients with grade I TCC, 65% with grade II TCC, 65% with grade III TCC, and 80% of patients with CIS. Immunoperoxidase labelling of CEA can enhance the sensitivity of exfoliative cytology.     

Expression of blood group antigens in bladder cancer: current concepts.

Blood group antigens are a group of carbohydrate structures bound to membrane lipids or proteins of erythrocytes and certain epithelial tissues including urothelium. The Lewis antigens are structures that are genetically and biochemically related to the ABO blood group. The ABO and Lewis blood group systems are differentially expressed in the normal urothelium of "secretors" versus "nonsector" individuals. The normal urothelium of "secretors" is rich ABH, Leb, and Ley antigens while the urothelium of "nonsecretors" does not express these antigens. Therefore, deletion of ABH antigens, commonly noted in TCC, can only be reliably ascertained in "secretor" individuals. Neoexpression of the Lewis X antigen (which is absent in normal urothelium) is noted in over 85% of TCC regardless of tumor stage and grade. Immunocytological detection of the Lewis X antigen on exfoliated bladder epithelial cells enhances the detection of urothelial tumor cells, particularly from low grade and low stage neoplasms.     

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。     

Precise microdissection of human bladder carcinomas reveals divergent tumor subclones in the same tumor

BACKGROUND: Human bladder carcinoma is thought to arise from a field change that affects the entire urothelium. Whether independently transformed urothelial cell populations exist in the same patient is uncertain. METHODS: We studied the clonality of urinary bladder carcinoma in 18 female patients who underwent cystectomy for urothelial carcinoma. None had multiple tumors. Tumor samples were obtained from different areas of the same tumor. Sixty-seven tumor samples were analyzed. Tumor genomic DNA was microdissected and extracted from formalin-fixed, paraffin-embedded slides. The clonality of urothelial tumors was evaluated on the basis of a polymorphism of the X chromosome-linked human androgen receptor gene (HUMARA) locus. The technique is dependent on digestion of DNA with the methylation-sensitive restriction enzyme HhaI, polymerase chain reaction (PCR) amplification of HUMARA locus, and detection of methylation of this locus. With this method, only the methylated HUMARA allele is selectively amplified by PCR. RESULTS: Eleven of 18 patients were informative. Nonrandom inactivation of the X chromosome was found in 9 of the 11 informative patients (82%). Seven patients showed different patterns of nonrandom X chromosome inactivation for tumor samples obtained from different regions of the same tumor. Two patients showed the same pattern of nonrandom X chromosome inactivation in all samples. CONCLUSIONS: Some muscle-invasive urothelial carcinomas may arise from independently transformed progenitor urothelial cells, supporting the "field effect" theory for bladder carcinogenesis.     

Role of human papillomavirus in the development of urothelial carcinoma

It has been estimated that almost 10% of the worldwide cancer burden is linked to human papillomavirus (HPV) infection. Although the association between HPV and bladder carcinoma has been extensively investigated, data on the role of HPV in bladder carcinogenesis are controversial. The aim of the study was to assess the possible role of human papillomavirus in the development of urothelial bladder carcinomas. Formalin-fixed and paraffin-embedded archival tissue samples were used for DNA extraction. Seventy urothelial bladder carcinoma tissues were screened by nested-polymerase chain reaction (PCR) for HPV DNA with a control group of total 18 cervical tissues with invasive cervical carcinoma and cervical intraepithelial neoplasia III (CIN III). In the study group, we did not find HPV DNA positivity in any of the urothelial carcinomas. In the control group, 15 out of 18 (83.3%) samples were positive for the HPV DNA. These results indicated that there was no association between HPV infection and urothelial carcinomas.     

血型抗原Lewis X免疫组化染色法与尿液脱落细胞学对膀胱移行细胞癌的诊断比较

目的:采用多中心临床试验方案,以病理诊断为金标准,比较尿液Lewis X抗原免疫组化染色法与尿液脱落细胞学(voided urine cytology,VUC)对膀胱移行细胞癌(TCC)的诊断价值。方法:受试对象为258例因怀疑膀胱癌收住入院的患者以及97例膀胱癌保留膀胱手术后随访患者。尿液脱落细胞标本分别行常规细胞学检查和免疫组化Envision二步法Lewis X抗原染色,计数每100个脱落细胞所含阳性染色细胞数目。结果:根据ROC曲线得出最佳阈值为5个阳性细胞/100脱落细胞,此时灵敏度单次试验为82.1%~84.2%,特异性为80.3%~78.9%,2次测定作为平行试验灵敏度为86.8%,特异性为77.0%;灵敏度提高,特异性略有下降。以11个阳性细胞/100脱落细胞为阈值的灵敏度和特异性分别为66.0%和95.4%。VUC的灵敏度和特异性分别为47.6%和94.7%。结论:Lewis X抗原灵敏度高于VUC,对低分级、早期肿瘤差别尤其明显。与B型超声结合可以提高灵敏度。     

Overexpression of epidermal growth factor receptor in urothelium elicits urothelial hyperplasia and promotes bladder tumor growth

Although urothelium is constantly bathed in high concentrations of epidermal growth factor (EGF) and most urothelial carcinomas overexpress EGF receptor (EGFr), relatively little is known about the role of EGFr signaling pathway in urothelial growth and transformation. In the present study, we used the uroplakin II gene promoter to drive the urothelial overexpression of EGFr in transgenic mice. Three transgenic lines were established, all expressing a higher level of the EGFr mRNA and protein in the urothelium than the nontransgenic controls. The overexpressed EGFr was functionally active because it was autophosphorylated, and its downstream mitogen-activated protein kinases were highly activated. Phenotypically, the urinary bladders of all transgenic lines developed simple urothelial hyperplasia that was strongly positive for proliferative cell nuclear antigen and weakly positive for bromodeoxyuridine incorporation. When coexpressed with the activated Ha-ras oncogene in double transgenic mice, EGFr had no apparent tumor-enhancing effects over the urothelial hyperplastic phenotype induced by Ha-ras oncogene. However, when coexpressed with the SV40 large T antigen, EGFr accelerated tumor growth and converted the carcinoma in situ of the SV40T mice into high-grade bladder carcinomas, without triggering tumor invasion. Our studies indicate that urothelial overexpression of EGFr can induce urothelial proliferation but not frank carcinoma formation. Our results also suggest that, whereas EGFr and Ha-ras, both of which act in the same signal transduction cascade, stimulated urothelial hyperplasia, they were not synergistic in urothelial tumorigenesis, and EGFr overexpression can cooperate with p53 and pRB dysfunction (as occurring in SV40T transgenic mice) to promote bladder tumor growth.     

Blood group ABO and Lewis antigen expression during neoplastic progression of human urothelium. Immunohistochemical study of type 1 chain structures

The deletion of blood group ABO antigen expression in bladder carcinoma has attracted attention because of its potential as a prognostic parameter. Based on recently produced monoclonal antibodies against blood group antigens, it has become possible to elucidate the carcinoma-associated modulation of these antigens at a molecular level. In this study we have used a panel of monoclonal antibodies (H, Lea, Leb, A, ALeb) that are specific to type 1 chain structures. By the use of an immunohistochemical method, the histologic and cytologic location of these antigens in the urothelium was studied in 25 biopsies from transitional cell carcinomas and compared to 21 previously examined normal biopsies. Urothelial blood group reactivity was compared to Lewis and secretor status. The authors found a series of events associated with neoplastic progression of noninvasive urothelium: a disruption of the orderly stratification of blood group antigens in different cell layers; cytostructural relocation of cytoplasmic antigens to the cell surface; loss of correlation between urothelial blood group antigens and secretor status; and gradual deletion of antigens. In the invasive tissue these events were followed by a total deletion of A and H isoantigens and uniform expression of Lewis b and sialyted Lewis a antigen. These findings indicate that there is a complex modulation of blood group antigen biosynthesis associated with the neoplastic progression of the human urothelium.     

IMP3 predicts aggressive superficial urothelial carcinoma of the bladder

PURPOSE: In this study, we investigated whether an oncofetal protein, IMP3, can serve as a new biomarker to predict progression and metastasis of early-stage urothelial carcinoma of the bladder. EXPERIMENTAL DESIGN: The expression of IMP3 in 242 patients with primary superficial bladder urothelial carcinoma and metastatic urothelial carcinoma was evaluated by immunohistochemistry. Patients with primary superficial urothelial carcinoma of the bladder were further investigated by use of survival analysis. RESULTS: Twenty percent (42 of 214) of primary superficial urothelial carcinomas and 93% (26 of 28) of metastatic urothelial carcinomas expressed IMP3. Kaplan-Meier plots and log-rank tests showed that patients with IMP3-positive tumors had a much lower progression-free survival (P = 0.0002) and disease-free survival rate (P = 0.0067) than did those with IMP3-negative tumors. The 5-year progression-free and disease-free survival rates were 91% and 94% in IMP3-negative patients versus 64% and 76% in IMP3-positive patients, respectively. Sixty percent of IMP3-positive patients with superficial invasive urothelial carcinoma at initial diagnosis went on to develop metastases, whereas no metastasis was found in IMP3-negative patients (P = 0.0017). In the multivariable Cox analysis, patients with IMP3 expression in their superficial urothelial carcinomas subsequently developed invasive tumors or metastasis at a rate that was about five times greater than cases without expression of IMP3 adjusting for other well-known clinical variables (tumor stage and grade, etc.). CONCLUSIONS: Our findings indicate that IMP3 is an independent prognostic marker that can identify a group of patients with a high potential to develop progression and who might benefit from early aggressive therapy.