膀胱癌端粒酶活性的研究

目的 :探讨端粒酶活性和膀胱癌之间的关系。方法 :采用在PCR基础上建立的TRAP ELISA法对30例膀胱癌及 9例切缘组织和 7例正常膀胱组织中端粒酶活性进行半定量的研究。结果 :膀胱癌组总阳性率为83.3% (2 5 / 30 ) ,与切缘组 11.1% (1/ 9)和对照组 0 .0 % (0 / 7)相比差异有极显著性意义 (P <0 .0 1) ;后两者之间差异无显著性意义 (P >0 .0 5 )。端粒酶阳性表达率和表达强度与患者年龄、性别、病变部位、肿瘤大小、手术方式等无显著性相关 (P >0 .0 5 ) ,而端粒酶表达强度与细胞病理分级具有相关性 (P <0 .0 5 )。结论 :膀胱肿瘤的端粒长度和端粒酶活性对于判断疾病的恶性程度、预后、监测微小残瘤病灶和预示早期复发均有积极的意义     

膀胱癌尿脱落细胞端粒酶活性检测及其临床意义

目的检测尿脱落细胞端粒酶活性并探讨其临床意义。方法应用改良的端粒重复序列扩增（ＴＲＡＰ）银染方法,分别对膀胱癌组织、正常膀胱组织,以及膀胱癌患者和非尿路上皮肿瘤患者的尿脱落细胞、膀胱冲洗液进行端粒酶活性检测。结果１２例正常膀胱组织均无端粒酶活性,４８例膀胱癌组织中４４例（９１．７％）端粒酶阳性。膀胱癌患者尿液及膀胱冲洗液中脱落细胞端粒酶阳性率分别为８３．３％（４０／４８）和８７．５％（４２／４８）。１２例分化良好（Ｇ１级）膀胱癌患者中,尿液和膀胱冲洗液中脱落细胞端粒酶阳性率分别为７５．０％（９／１２）和８３．３％（１０／１２）。结论尿脱落细胞端粒酶活性检测敏感性高,可用于膀胱癌的早期诊断和术后随访。     

Telomerase activity in bilharzial bladder cancer: Prognostic implications

Background: Bladder cancer is a common malignancy in Egypt and other developing countries in which infection with Schistosoma haematobium is prevalent. Bladder cancer caused by bilharziasis has different clinical and biological characters than that observed in the western world. In this study, we used the TRAP technique to estimate telomerase activity in bilharzial bladder cancer specimens and we correlated the findings with other clinical and pathological findings. Patients and methods: Bladder cancer specimens were obtained from 57 patients who underwent radical cystectomy and pathological diagnosis was obtained in all patients. Tissue samples were frozen in liquid nitrogen and stored at −80°C. Telomerase activity by PCR-ELISA technique was measured using TRAP technique. Results: Our patient group included 45 males and 12 females with a median age of 49 years. The majority of our patients (35/57) have squamous histology and they have proven bilharzial history shown in the pathology specimens. Stage P3b was encountered in 29/57 patients whereas thirty-five patients have grade II tumors. The majority of our patients (41/57) were negative for pelvic nodes metastases. Telomerase activity was detected in 27/57 patients (47.4%). The mean level of telomerase was 0.85±0.77 in positive patients and 0.029±0.025 in negative patients. The expression of telomerase and its mean level in patients above age of 60, in males and in those with squamous pathology, higher grade of tumors or positive node was higher than those without but the difference did not reach statistical significance (P>0.05). Alternatively, expression was significantly higher in those with stages (P1–P3a) compared with P3b–P4a disease stages (66.6% vs. 37.1, P=0.03). Conclusion: Telomerase activity is increased in bilharzial bladder cancer although to a lesser degree than that reported for TCC in the western world, which could be explained, by different biological behavior or different assay methods. Further larger studies with more number of patients are still needed to determine its potential value for early detection and possible use as a therapeutic target.     

膀胱癌组织端粒酶活性的研究

目的　探讨膀胱癌组织端粒酶活性的临床意义。　方法　应用银染端粒重复序列扩增法检测 42例膀胱癌及其癌旁组织的端粒酶活性。　结果　 3例正常膀胱组织端粒酶表达均阴性 ;42例膀胱癌组织中端粒酶表达阳性 35例 (83 .3 % ) ,癌旁组织端粒酶表达阳性 7例 (16 .7% ) ;浸润癌或有淋巴结或远处转移者端粒酶表达阳性率高于非浸润癌或无转移者 ,但差别无显著性意义 (P >0 .0 5 )。　结论　端粒酶是膀胱癌较理想的肿瘤标记物之一。     

Telomerase in human bladder cancer

Biomarkers for human bladder cancer are currently available and more are being developed. However, the ultimate goal of diagnosing bladder cancer consistently in a noninvasive fashion has not yet been achieved. Telomerase is an enzyme that may play a role in maintaining telomere sequences in the ends of chromosomes and its activity may reflect the presence of immortal or cancer cells. In this article, we reviewed the potential applications of telomerase in the diagnosis, monitoring, and treatment of human bladder cancer.     

膀胱肿瘤端粒酶活性及其对预后的影响

目的：探讨瑞粒酶活性与膀胱癌临床生物学行为及预后的关系。方法：采用ＴＲＡＰ法４６例膀胱癌组织端粒酶提取液以及１０倍、１００倍稀释液分别进行端粒酶活性检测并结合临床资料进行分析。结果：４６例膀胱癌组织有３８例（８２．６％）端粒酶活性表达阳性。端粒酶活性强度与病人年龄、性别、肿瘤大小、数目无关（Ｐ均＞０．０５）;而Ｇ３肿瘤端粒酶活性强度较Ｇ２和Ｇ１肿瘤明显增高,Ｐ＜０．０１;浸润性肿瘤（Ｔ２－４）较浅     

膀胱癌癌旁组织端粒酶活性检测的临床意义

目的　探讨膀胱癌癌旁组织端粒酶活性检测的临床意义。　方法　采用端粒重复序列扩增 (TRAP)法 ,检测 2 4例膀胱癌组织及癌旁组织中端粒酶活性表达。　结果　 2 4例癌旁组织中端粒酶活性表达阳性 10例 (42 % ) ,癌旁组织端粒酶活性表达与原发灶癌病理分级分期相关 ,并与癌复发相关。　结论　膀胱癌癌旁组织端粒酶活性检测可作为判断膀胱癌预后的指标之一     

胰腺癌组织中端粒酶活性的表达

目的　探讨端粒酶活性与胰腺癌之间的关系 ,评价其作为胰腺癌诊断新的肿瘤标志物的可能性。方法　采用重复片段扩增SYBRGreen染色法 ,对 42例胰腺癌癌患者癌组织及其癌旁组织的端粒酶活性进行检测。结果　胰腺癌组织中端粒酶阳性率为 80 .9% (3 4/4 2 ) ,而在癌旁组织中仅 7.1% (3 /4 2 )表达端粒酶活性 ,胰腺癌组织端粒酶活性显著高于癌旁组织 (P <0 .0 0 1)。端粒酶活性的表达与胰腺癌组织的肿瘤大小、淋巴结转移、病理学临床分期及分化程度相关。结论端粒酶活性是特异性较强的恶性肿瘤分子标志物 ,其检测有可能成为胰腺癌诊断和预后判断的有效指标     

Telomerase activity in urine after transurethral resection of superficial bladder cancer and early recurrence

BACKGROUND: To explore the relationship between telomerase activity in urine after transurethral resection (TUR) of superficial bladder cancer and early intravesical recurrence. METHODS: Urine samples were obtained from 42 patients with superficial bladder cancers prior to TUR and on the postoperative day 1 and day 6. These patients were followed-up prospectively by cystoscopy at 3 and 6 months after TUR in combination with urinary cytology and telomerase activity. Telomerase activity in the urine was assessed by the telomeric repeat amplification protocol assay. RESULTS: Urinary telomerase activity prior to TUR was positive in 24 (57%) of the 42 patients. On the postoperative day 1 and day 6, positive urinary telomerase activity was seen in 13 (31%) and nine (21%) patients, respectively. Postoperative urinary telomerase activity on either day 1 or day 6 was significantly associated with pre-operative urinary telomerase activity status (P = 0.0024). Fifteen patients showed intravesical tumor recurrence at 3 months cystoscopic check-up and an additional nine had recurred at the 6 months check-up. Recurrence rate within 6 months in patients with pre-operative positive urinary telomerase activity was similar to that in those with negative activity (58.3 vs 58.8%). However, recurrence rate at 3 months for patients with positive activity was higher than that of those with negative activity (50 vs 17.7%), in 23 patients treated only by TUR. CONCLUSIONS: Presence of cells positive for telomerase activity in urine after TUR of superficial bladder cancer indicates persistently existing cancer cells in the urine. It is, however, not a sole predictor of the early intravesical recurrence.     

膀胱肿瘤端粒酶活性及细胞凋亡的研究

目的　探讨端粒酶活性及细胞凋亡与膀胱癌临床生物学行为及预后的关系。　方法　分别采用端粒酶活性试剂盒和末端脱氧核苷酸转移酶介导缺口末端标记 (TUNEL)法对 5 6例膀胱癌标本进行端粒酶活性和细胞凋亡的检测 ,并结合临床资料进行分析。　结果　 5 6例膀胱癌标本端粒酶活性阳性率为 89 3 % (5 0 / 5 6 ) ,细胞凋亡指数为 (4 5 2± 14 4) % ;端粒酶活性强度及细胞凋亡指数与年龄、性别、肿瘤大小、数目等无关 (P >0 0 5 ) ;与肿瘤分级、分期及预后有关 (P <0 0 1) ,即端粒酶活性越高或 (和 )细胞凋亡越少 ,则肿瘤“瘤级”、“瘤期”高者预后较差。端粒酶活性强度与细胞凋亡指数呈明显的负相关 (r=- 0 6 9,P <0 0 1)。　结论　膀胱癌端粒酶活性及细胞凋亡与肿瘤分级、分期及预后有关 ,端粒酶活性及细胞凋亡的检测有助于膀胱癌的临床分析及预后的评估     

尿脱落细胞端粒酶活性表达对尿路上皮癌的诊断价值

目的：探讨尿脱落细胞端粒酶活性表达对尿路上皮癌的诊断价值。方法：采用ＰＣＲ－ＣＬＩＳＡ法检测尿液中脱落的肿瘤细胞端粒酶活性,并以肾癌及非肿瘤患者的尿标本对照。结果：５９例尿路上皮癌患者尿脱落细胞端粒酶活性表达率为８８．１％,而肾癌患者为１０．０％,非肿瘤患者为２０．０％,前者与后两者比较,有极显著性差异（Ｐ〈０．０１）;不同分化程度及不同临床分期膀胱癌患者尿脱落细胞端粒酶活性表达率无显著性差异（Ｐ     

Telomerase activity as a potential marker in preneoplastic bladder lesions

OBJECTIVE: To assess telomerase activity (involved in cell immortalization and detectable in most malignant tumours but not in normal somatic tissues) as a marker in cancer diagnosis. PATIENTS AND METHODS: Tissue telomerase activity was assayed by two different techniques, the telomeric repeat amplification protocol-polymerase chain reaction (TRAP-PCR) and a telomerase PCR-enzyme linked immunosorbent assay. Malignant and inflammatory bladder lesions and their adjacent normal tissues were assessed for telomerase activity in a group of 18 patients, 14 of whom had urothelial carcinoma and four a nonspecific inflammatory lesion of the bladder. RESULTS: Eleven of the 14 tumour samples analysed were telomerase-positive and two of the three telomerase-negative tumour samples had a detectable 'telomerase inhibitor'. In the apparently normal tissues next to bladder tumours, four of the 14 specimens were telomerase-positive. Interestingly, these lesions were always next to high-grade muscle-invasive bladder tumours (pT2G3). Two of the four nonspecific inflammatory lesions (one of cystitis glandularis and one of severe dysplasia), known to be preneoplastic lesions, were also telomerase-positive. CONCLUSION: These results strongly suggest that the reactivation of telomerase may be an early event in bladder carcinogenesis, preceding morphological changes related to malignant transformation. Telomerase activity may therefore be useful both as an indicator of malignant potential in preneoplastic lesions, e.g. cystitis glandularis and severe dysplasia, and as a prognostic marker of bladder tumour relapse or progression.     

Telomerase activity in disseminated prostate cancer cells

OBJECTIVE: To analyse telomerase activity in disseminated prostate cancer cells isolated from bone marrow aspirates taken from men with localized prostate cancer before radical prostatectomy (RP). PATIENTS AND METHODS: Disseminated epithelial prostate cancer cells were isolated from bone marrow aspirates from 69 men with localized prostate cancer before RP, by magnetic column-chromatography enrichment, followed by isolation of fluorescently labelled epithelial cells by micropipetting. We used pools of 10 non-epithelial bone marrow cells after tumour cell enrichment as control samples. These pure cell pools were tested for the presence of telomerase activity. RESULTS: In all, 49 of the patient samples contained disseminated prostate cancer cells. Homogeneous pools of 10 cells were obtained from 35 of these; 49% of the 35 specimens showed telomerase activity, whereas all five control samples did not. Telomerase activity in the 35 samples was not significantly associated with Gleason score, preoperative prostate-specific antigen level, tumour stage, or surgical margin status. Follow-up is continuing to assess an association with disease recurrence. CONCLUSION: This work shows the feasibility of isolating disseminated cancer cells for analysing individual or pooled cells. Compared to tissue staining, where telomerase is detected in 80-90% of samples, we found lower rates of telomerase activity in the disseminated tumour cells (49%). Telomerase-negative cells might provide information about cell dormancy, as telomerase is a marker of cell proliferation in immortal and cancer cells. Telomerase-positive cells might predict early disease recurrence, but a longer follow-up is needed to test this possibility.     

胃癌与端粒酶活性表达及DNA倍体关系的研究

目的　揭示胃癌与端粒酶活性及DNA倍体的关系。方法　检测 30例胃癌标本 ,同时取无瘤残端作为对照。端粒酶检测采用端粒重复扩增 酶联免疫吸附法 (TRAP ELISA法 )。DNA倍体的测定采用流式细胞术 ,一步法检测DNA含量。结果　肿瘤瘤体端粒酶阳性率 83 3% (2 5 / 30 ) ,无瘤残端端粒酶阳性率 3 3%(1/ 30 ) (P <0 .0 5 ) ;端粒酶阳性瘤体平均直径 6 5cm ,阴性瘤体平均直径 3 6cm(P <0 .0 5 ) ;端粒酶阳性肿瘤淋巴转移率 5 3 3% (16 / 30 ) ,阴性者无淋巴转移 (0 / 5 ) (P <0 .0 5 ) ;端粒酶阳性肿瘤中异倍体肿瘤占 5 6 0 % (14/2 5 ) ,而端粒酶阴性者无异倍体出现 (P <0 .0 5 )。结论　胃癌端粒酶活性升高 ,端粒酶阳性肿瘤瘤体大 ,淋巴转移率高 ,且异倍体发生率高 ,预后差。提示端粒酶的激活与胃癌的发生发展有密切关系。     

大肠癌患者粪便标本的端粒酶活性研究

目的 探讨通过粪便途径筛查大肠癌的可行性和新方法。方法 应用聚合酶链端粒重复扩增（PCR－TRAP）银染技术,研究了43例大肠癌患者粪便中脱落细胞的端粒酶活性表达。结果 62．8％的大肠癌患者粪便标本中有端粒酶阳性表达。患者粪便标本中端粒酶阳性表达与其大肠癌Dukes分期、淋巴结转移和癌肿部位未见显著相关（P＞0．05）。1例结肠腺瘤患者粪便标本端粒酶表达阳性,其腺瘤组织也存在端粒酶活性表达。粪便端粒酶检测的敏感性、特异性和阳性预测值分别为62．8％、95．7％和96．4％。结论 PCR－TRAP银染检测大肠癌患者粪便脱落细胞的端粒酶活性表达为改善大肠癌筛查方法和大肠癌诊断作了新的尝试。     

Tumor-adopted diagnosis of bladder cancer

70% of our patients suffering from bladder cancer present themselves initially with a superficial tumor (Tis, Ta, T1) and only 30% are initially seen with a muscle-invasive carcinoma (T2, T3-4, N+, M+). Clinical history, physical examination, urine cytology, IVP and cystoscopy in combination with adequate tissue harvest by transurethral resection are the basis of our diagnostic procedures. Bimanual palpation, systematic biopsies, sonography, computed tomography, bone scan and further procedures are not to be considered as routine examinations and are only used in special situations. Only exactly defined diagnostic algorithms permit the development and use of valid prognostic parameters. They further allow to perform a tumor-orientated therapy and to compare patient groups which have been treated in different institutions according to similar or different therapeutic regimens. This includes the importance of a TNM-oriented therapy. It has to be stressed though that the prevalence of distinct pathological changes, such as bone metastases in T1 tumors, as well as financial resources, have to be taken into account. Further, definite therapeutic intention such as a curative vs. palliative regimen is a decisive criterium for the amount of performed diagnostic procedures. It also is of importance that new diagnostic modalities, if introduced into the clinical routine, have to be investigated concerning their validity in relevant and large patient cohorts and their rationale in our diagnostic algorithm has to be defined. This is predominantly not the case in radiological imaging techniques and thus a large amount of resources are wasted. Copyright Copyright 1999 S. Karger AG, Basel     

Computer-assisted cystoscopy diagnosis of bladder cancer

Objectives

One of the most reliable methods for diagnosing bladder cancer is cystoscopy. Depending on the findings, this may be followed by a referral to a more experienced urologist or a biopsy and histological analysis of suspicious lesion. In this work, we explore whether computer-assisted triage of cystoscopy findings can identify low-risk lesions and reduce the number of referrals or biopsies, associated complications, and costs, although reducing subjectivity of the procedure and indicating when the risk of a lesion being malignant is minimal.