Occurrence of chromosome 9 and p53 alterations in multifocal dysplasia and carcinoma in situ of human urinary bladder

To define the genetic changes of flat urothelial lesions, carcinoma in situ (CIS) and moderate dysplasias (DII) were investigated for alterations in the two chromosomal regions most frequently involved in bladder cancer. Overall, 33 CIS and 16 DII from 21 patients were used to microdissect urothelium. Dual color fluorescence in situ hybridization (FISH) using gene locus probes of 9q22 (FACC), 9p21 (CDK), 17p13 (p53), and related centromeric probes was applied on interphase nuclei. In parallel, preamplified DNA of these samples was used for loss of heterozygosity (LOH) analyses with eight microsatellite markers on chromosomes 9p, 9q and 17p, and for sequencing of exons 5-9 of p53. Data indicated nearly identical deletion frequencies for chromosomes 9 and 17 for CIS (chromosome 9, 86%; p53, 84%). DII showed a lower deletion rate in comparison with CIS (chromosome 9, 75%; p53, 53%). A very high correlation between the results of FISH and LOH analyses was found. p53 mutations were detected in 12 of 15 patients (CIS, 72%; DII, 67%). In three of 16 patients with multifocal tumors, oligoclonal lesions were identified by LOH analyses, a finding further supported by sequencing of p53, by which two different p53 deletions were detected in two cases. In conclusion, data from microdissected flat urothelial lesions indicate that chromosome 9 deletions cannot be regarded as indicators of papillary growth, because they are found frequently in both types of flat lesions of the urothelium: those associated with papillary tumors and those that are not. The similar distribution and lower amount of genetic changes in DII render DII a possible precursor lesion of CIS.     

Viability of the human cervical epithelium with dysplasia and carcinoma in situ

Autoradiograms of histologic slides of 58 human cone specimens with dysplasia and carcinoma in situ were analyzed after tissue samples were incubated with labeled RNA precursors. Both the vertical and lateral distributions of labeled cells were rather uniform in the major part of the epithelium, which suggested that the tissue remained metabolically active during incubation. Only the uppermost epithelial cells in heavily labeled areas were devitalized as deduced by the morphologic appearance of the cells, the absence of labeling in the cells, the trypan blue exclusion test, and the trypsin digestion test. The viability of large epithelial areas suggested that the previously reported focal distribution of proliferating and nonproliferating areas in the cervical epithelium is a genuine phenomenon and not the result of focal epithelial devitalization acquired during incubation.     

Whole bladder wall photodynamic therapy for refractory carcinoma in situ of the bladder.

Whole bladder wall photodynamic therapy (PDT) with haematoporphyrin derivative and an argon dye laser as a light source was performed on 34 patients with refractory carcinoma in situ (CIS) of the bladder. Twenty-five of the 34 patients (73.5%) had achieved a complete response (CR) at 3 months after the treatment. The median follow-up for these CR patients is 49.3 months. Although recurrence within 2 years of follow-up occurred in 14 (77.8%) of the 18 CR patients followed to that point, since most of the recurrent tumours were superficial and low-grade papillary tumours, transurethral resection of the bladder tumours appeared to be sufficient. Of the total of 34 patients, ten were alive with bladder intact with a mean follow-up period of 64.0 months. Skin photosensitivity and transient decrease in bladder capacity were noted as adverse reactions, caused by retention of haematoporphyrin derivative in the skin and normal portion of the bladder. These data suggest that PDT can be an effective form of therapy for CIS of the bladder.     

Expression of blood-group-related antigens in carcinoma in situ of the urinary bladder

Expression of epithelial ABH blood group antigens and the T (Thomsen-Friedenreich)-antigen was quantitatively studied by immunoperoxidase techniques in nine cystectomy specimens containing extensive carcinoma in situ (CIS), and also histologically benign epithelium. CIS areas typically showed abnormal expression of both ABH and T-antigens and a distinctive vascular architecture, revealed by endothelial ABH staining. Histologically normal epithelium generally was antigenically normal, but occasionally showed abnormalities of either ABH or T-antigens. Antigen expression was variable in histologically atypical epithelium, with significant segments showing abnormalities of either ABH or T-antigen, and sometimes of both antigenic markers. It is postulated that histologically benign, but antigenically abnormal, epithelium may represent low-grade CIS of the urinary bladder. Assessment of blood-group-related antigen expression in flat atypical epithelium of the urinary bladder may be useful for predicting the biologic potential of these lesions.     

Survival of patients with carcinoma in situ of the urinary bladder.

BACKGROUND: To the authors' knowledge, the long term follow-up of patients with carcinoma in situ of the urinary bladder is limited. METHODS: The authors studied 138 patients diagnosed with urothelial carcinoma in situ of the bladder at the Mayo Clinic between 1972-1979. All the histologic slides were reviewed and fulfilled the diagnostic criteria for carcinoma in situ according to the newly proposed World Health Organization and International Society of Urologic Pathology classification system. None of these patients had previous or coexisting invasive urothelial carcinoma at the time of diagnosis. Cox proportional hazards models were used to determine the prognostic significance of numerous clinical and pathologic findings using progression free, cancer specific, and all-cause survival as the endpoints for analysis. Progression was defined as the development of invasive carcinoma, distant metastases, or death from bladder carcinoma. RESULTS: The patients ages at the time of diagnosis ranged from 32-90 years (mean, 65.6 years). The male to female ratio was 7:1. Carcinoma in situ usually was multifocal (50%) with a predilection for the trigone, lateral wall, and dome. The mean follow-up after surgery was 11.0 years (range, 0.7-25 years). Actuarial progression free, cancer specific, and all-cause survival rates were 63%, 79%, and 55%, respectively, at 10 years, and 59%, 74%, and 40%, respectively, at 15 years. The mean interval from the time of diagnosis to cancer progression was 5 years. Patient age at diagnosis was significant in predicting progression free (P = 0.01) and all-cause survival (P = 0.002). Cystectomy performed within 3 months after the initial diagnosis was associated with improved all-cause survival (P = 0.03). After controlling for age, there was no difference in survival between patients who received immediate cystectomy and those did not (P = 0.16). CONCLUSIONS: Patients with carcinoma in situ of the bladder are at significant risk of cancer progression and death from bladder carcinoma. Cystectomy does not appear to offer a significant survival advantage in patients with carcinoma in situ of the bladder after adjusting for age.     

Urothelial papilloma of the bladder. Clinical and biologic implications

BACKGROUND: An international consensus has been reached regarding diagnostic criteria for papilloma of the urinary bladder. However, the incidences of recurrence and progression in patients with urothelial papilloma are uncertain. METHODS: The population for this study consisted of 52 patients who were diagnosed with urothelial papilloma of the bladder at the Mayo Clinic between 1914 and 1998. All histologic slides were reviewed and fulfilled the diagnostic criteria of urothelial papilloma from the 1998 World Health Organization/International Society of Urological Pathology classification system. No patients had previous or coexistent urothelial carcinoma, and none were treated after biopsy. RESULTS: The mean patient age at diagnosis was 57 years (range, 22-89 years). The male-to-female ratio was 1.9:1. The mean follow-up was 9.8 years (range, 0.1-58 years). Four patients developed recurrent papilloma (mean interval from diagnosis to recurrence, 3.3 years); 1 other patient developed papillary neoplasm of low malignant potential (Ta WHO Grade 1 papillary urothelial carcinoma) 6 years after the initial diagnosis of papilloma. None of these patients developed dysplasia, carcinoma in situ, or invasive urothelial carcinoma or died of bladder cancer. CONCLUSIONS: Patients with urothelial papilloma have a low incidence of recurrence and rarely, if ever, develop urothelial carcinoma.     

Characteristics of women with dysplasia or carcinoma in situ of the cervix uteri.

To identify risk factors for various cervical abnormalities, 237 women with abnoromal cervical smears and 422 control women were interviewed. Cervical biopsy specimens taken from the patients with abnormal smears were reviewed according to standard criteria by one pathologist and classified as follows: 65 carcinoma in situ, 81 severe dysplasia, 44 mild dysplasia and 47 normal histology. Factors associated with risk of mild dysplasia, severe dysplasia and carcinoma in situ were similar to those previously identified for invasive carcinoma, and included age at first intercourse, multiple sexual partners and pregnancy outside marriage. Analysis to disentangle correlated factors revealed that number of sexual partners exerted effects independently of age at first intercourse, whereas the reverse was not true. This finding fails to support suggestions that adolescence is a period when the cervix is most vulnerable to the effects of sexual behaviour. Other factors relating to risk of cervical abnormalities were smoking and use of oral contraceptives. It was not possible to show that these relationships were incidental, but further investigation is required to establish whether they are causal.     

Blood group isoantigen deletion in carcinoma in situ of the urinary bladder.

Blood group isoantigens A and H (O) were measured by the Secific Red Cell Adherence (SRCA) Test in nine radical cystectomy specimens removed from patients with extensive carcinoma in situ of the urinary bladder. All bladders had areas of histologically normal epithelium and areas of epithelial atypia in addition to the carcinoma in situ. In eight cases, tissue-associated blood group isoantigens were deleted in areas showing either atypia or carcinoma in situ. Isoantigens were present in these areas in the ninth case. Blood group isoantigens were absent in approximately half of the sections of histologically normal epithelium. We propose that areas of epithelium which appear benign according to conventional histological criteria but in which the blood group isoantigens are absent may represent areas of low grade carcinoma in situ.     

荧光原位杂交在尿路上皮癌诊断中的应用

目的：检测尿路上皮肿瘤患者尿液脱落细胞染色体的缺失和非整倍异常,探讨FISH技术作为尿路上皮肿瘤患者无创诊断方法的价值。方法：收集可疑尿路上皮肿瘤患者和健康对照人群的新鲜尿液,同步进行细胞形态学分析及荧光原位杂交（Fluorescence in situ hybridization, FISH） 检测3号、7号及17号染色体、9号染色体p16位点异常。共入选可疑尿路上皮肿瘤患者100例,正常健康对照组20例,采用正常对照组患者各染色体异常数据设定阈值用于肿瘤患者的实验室诊断。根据检验结果与病理结果对照分别计算FISH和脱落细胞的敏感度和特异度并进行统计学分析。结果：与正常对照组相比,尿路上皮肿瘤患者尿液脱落细胞染色体异常明显增多。尿脱落细胞学的敏感度和特异度分别为71%和80%,FISH的敏感度和特异度分别为88%和80%（P<0.01）。根据两种检测方法的敏感度和特异度绘制的接受者工作特征（ROC）曲线显示尿脱落细胞学和FISH的曲线下面积分别为0.758和0.842。结论：对可疑尿路上皮肿瘤的患者进行FISH检测是一种有价值的无创检测方法。FISH的总体敏感度高于尿脱落细胞学,特异度与尿脱落细胞学相当。     

多色荧光原位杂交在膀胱尿路I皮癌诊断中的应用

背景与目的:膀胱尿路上皮癌是我国泌尿生殖系统常见的恶性肿瘤.本研究旨在分析中国人膀胱尿路上皮癌中染色体畸变的情况,探讨多色荧光原位杂交(multicolor fluorescence in situ hybridization,M-FISH)技术辅助诊断膀胱尿路上皮癌的可行性和有效性.方法:用随机引物法标记3、7、17号染色体着丝粒及9p21区带探针,对57例膀胱尿路上皮癌间期细胞核进行荧光原位杂交(fluorescence in situ hybridization,FISH),统计染色体畸变情况并分析其与病理分期、分级的关系以及染色体畸变组合诊断膀胱尿路上皮癌的阳性率.结果:3、7、17号染色体和9p21的畸变率分别为47.4%(27/57)、50.9%(29/57)、56.1%(32/57)和59.6%(34/57),畸变与分期无相关性,3、7和17号染色体畸变与病理分级有显著相关性(P＜0.01).四个探针组合诊断膀胱尿路上皮癌的总阳性率为54.4%.结论:M-FISH技术检测有助于探索3、7、17号染色体畸变与病理分级的关系.     

Use of fluorescent in situ hybridization in the cytogenetic diagnosis of urinary bladder urothelial carcinoma

Our study was concerned with assessing the results of FISH analysis as a procedure to detect occult features of tumor and contribute to individualizing treatment strategies. It included a cohort of 50 patients with primary bladder tumors who had undergone transurethral resection. Washings from all patients were tested using UroVysion protocol immediately prior to surgery. Patients were divided into groups according to stage and grade of tumor. The sensitivities were 81.5%, 91.7% and 100% for Ta, T1 and T2 groups, respectively. In G1, G2 and G3 groups, the sensitivities were 70%, 100% and 100%, respectively. Besides, the rate of detecting genetically abnormal cells was significantly higher at stage T2 as compared with Ta and Ta+T1 groups. It was also higher in G3 group as compared with G1 for Ta and G2. No significant differences in the mean number of signals from each chromosome was reported. Frequency rates of abnormal cells of less than 40% correlated with absence of invasion into the muscular layer and poorly-differentiated cell status. They rose insignificantly in proportion to stage and grade. Detection of less than 40% of genetically abnormal cells correlated with absence of mascular invasion and G3 tumor (90% credibility). FISH diagnostic procedure showed high sensitivity of detecting even at incipient stage of bladder carcinoma and was instrumental in preoperative predicting type of tumor.     

E-cadherin promotes intraepithelial expansion of bladder carcinoma cells in an in vitro model of carcinoma in situ

High-grade transitional cell carcinomas (TCCs) of the urinary bladder are frequently associated with carcinoma in situ, which may replace large areas of the mucosa of the urinary tract. The invasive component of TCCs often reveals a loss of expression of the cell-cell adhesion molecule E-cadherin, but the role of E-cadherin in the development and expansion of intraepithelial neoplasia is unknown. To study the underlying mechanism of intraepithelial expansion (IEE), we have developed an IEE assay. Human TCC cell lines were investigated in this IEE assay for their capacity to replace the surrounding normal murine urothelial cells. In vitro IEE appeared to be prominent in three (SD, RT112, and 1207) of the four E-cadherin-positive cell lines. Although the two E-cadherin-negative cell lines (T24 and J82) were able to penetrate surrounding normal urothelium as single cells, they largely lacked the capacity of IEE. These results prompted us to investigate whether the cell-cell adhesion molecule E-cadherin is an important determinant for IEE. T24 cells that were transfected with full-length mouse E-cadherin cDNA displayed an enhanced IEE rate. Transfection did not influence their proliferative capacity, their pattern and level of integrin expression, or their ability to expand in the absence of surrounding urothelium. The data suggest that E-cadherin-mediated cohesiveness is an important factor in the IEE of bladder carcinoma cells. These observations argue for a dual, paradoxical role of E-cadherin in bladder tumorigenesis. On the one hand, E-cadherin promotes the expansion of intraepithelial neoplasia; on the other hand, its loss correlates with invasive behavior.     

Urothelial tumors of the urinary bladder in manipur: a histopathological perspective

Objective: To study the histomorphological pattern of urothelial tumors of the urinary bladder in Manipurand to evaluate whether any correlation exists between tumor grade and muscle invasion. Methods: A 10 yearretrospective study of all consecutive cases diagnosed in the Department of Pathology RIMS – Imphal, between1st January 2001 to 31st December 2010. Results: The study included 26 cases of transitional cell tumors ofurinary bladder. The male to female ratio was 1.5: 1 and the ages ranged from 38 years to 73 years (mediansof 60 and 64 years, respectively). Of the total, 14 (53.9%) cases were low grade, 9 (34.6%) were high grade, 2(7.7%) were papillomas and 1 (3.9%) was a papillary urothelial neoplasm of low malignant potential (PUNLMP).Pathological staging showed that 14 (53.9%) of the cases were stage PTa, four (15.4%) PT1, and eight (30.9%)PT2. Some 18.2% of low grade tumors and 75% of high grade tumors were invasive to the detrusor musclelayer. Conclusion: Bladder cancer is an uncommon disease, transitional tumors being the only histological typeobserved. It was more common in males than females, with peak incidence in seventh decade. Most of the tumorswere non- invasive (PTa) and invasion to the detrusor muscle layer was seen in more than half of the high gradetumors. There is a definite correlation between advancing tumor grade and muscle invasion.