Telomerase activity in touch-imprint cell preparations from fresh prostate needle biopsy specimens.

OBJECTIVE: To evaluate whether it is possible to detect telomerase activity in cells exfoliated from prostate biopsies immediately before fixation. METHODS: A total of 115 transrectal biopsies of prostate tissue from 49 patients were touch-imprinted on an RNase-free microscope slide and then fixed. Touch imprints were immediately frozen and used to extract telomerase. Telomerase activity was determined by a telomeric repeat amplification protocol (TRAP) using a PCR-ELISA method. Inflammation and epithelial cells in each biopsy were quantitated by image cytometry. RESULTS: A total of 90/115 extracts had a proteic content suitable for analysis. Telomerase activity was detected in 18/26 (70%) carcinomas, 2/9 (22%) low-grade prostatic intraepithelial neoplasia (PIN) lesions, and 1/3 (33%) high-grade PIN lesions. In 4 of 7 patients with telomerase-positive tumors, telomerase activity was also found in a distant site devoid of morphologically detectable cancer cells. Telomerase activity was detected in touch imprints from fragments with less than 1 mm(2) of epithelial tissue, and was not associated with the extent of inflammation. CONCLUSIONS: From the technical stand point, the touch-imprint method may provide a useful adjunct for telomerase detection in prostate biopsies. With this procedure the bioptic fragment is left intact for histological examination. Diagnostically, the presence of telomerase activity in sites distant from the original tumor might suggest the presence of tumor cells that are morphologically undetectable.     

前列腺穿刺活检组织端粒酶活性检测及其临床意义

通过检测国人前列腺穿刺活检组织端粒酶活性,探讨其在前列腺癌诊断和预后的临床意义。20例前列腺癌标本和16例癌旁组织均来自前列腺穿刺活检,14例良性前列腺增生(BPH)标本取自前列腺摘除手术,均经病理证实。采用改良的TRAP-银染方法检测端粒酶活性,并进行半定量分析。结果20份前列腺癌标本中18份测得端粒酶活性(90％)。在16例癌旁前列腺组织中,64％(7/11)的前列腺上皮肉瘤(PIN)及40％(2/5)的癌旁BPH组织有端粒酶活性表达。14例BPH标本端粒酶活性均为阴性。18例前列腺癌阳性标本的端粒酶活性强度与肿瘤分级分期及PSA水平呈正相关。前列腺穿刺活检组织端粒酶活性检测可作为前列腺癌诊断的一项敏感性指标,其在前列腺癌预后中的价值有待于进一步的研究。     

前列腺组织端粒酶活性检测对前列腺癌的诊断价值

目的 通过检测前列腺穿刺组织端粒酶活性,探讨其在前列腺癌诊断和预后中的临床价值。方法 ２０例前列腺癌标本和１６例癌旁组织均来自前列腺穿刺活检,１４例良隆前列腺增生（ＢＰＨ）标本取自前列腺摘除手术,均经病理证实。采用改良的ＴＲＡＰ－银染方法检测端粒酶活性,并进行半定量分析。结果 ２０例前列腺癌标本中１８例测得端粒酶活性。１６例前列腺癌旁组织中,７例前列腺上皮内瘤（ＰＩＮ）,２例癌旁ＢＰＨ组织有端粒酶     

Detection of telomerase activity in prostatic fluid specimens

We report here the extended use of the telomeric repeat amplification protocol (TRAP) assay for the detection of telomerase activity in fresh prostatic fluid obtained from anesthetized patients. Telomerase activity was detected in pellet extract and/or supernatant fluid of specimens obtained from 25 of 30 prostate cancer (PCa) patients (83%), whereas no activity was similarly detectable in specimens taken from 8 of 9 patients (89%) without clinical evidence of PCa. The positive predictive value (PPV) of the TRAP assay for PCa in this pilot study was 96%. We found a strong correlation between telomerase activity in prostatic fluid specimens and serum prostate specific antigen (PSA) values. Telomerase activity was found in 84% of specimens from patients with PSA values >4 ng/ml, whereas in specimens from patients with PSA values 4. In prostatic fluid from PCa patients with Gleason scores of 相似文献   

Telomerase activity in prostate sextant needle cores from radical prostatectomy specimens

Human telomerase acts to maintain functioning telomeres, which are required for cellular immortality and very likely for cancer progression. Telomerase activity is present in about 85% of human cancers tested, but it has not been found in most human somatic cells and tissues. We used the Telomeric Repeat Amplification Protocol to perform telomerase activity assays on sextant needle core samples obtained from 35 freshly excised radical retropubic prostatectomy specimens. Similar assays were done on prostatic tissues obtained by means of other urologic procedures from 8 patients without prostate cancer. Telomerase activity was found in one or more specimens from 32 of 35 prostate cancer patients (91%), but was not detectable in all biopsy specimens from 7 of 8 cancer-free patients (88%). Further analysis showed that cancers more poorly differentiated, with higher Gleason scores, were always associated with a higher rate of telomerase detection and stronger telomerase activity. Moreover, comparison of telomerase activity in needle core samples with the volume of cancer in surrounding tissue as observed on corresponding histologic slides showed that stronger activity was positively correlated with a higher cancer volume. Prognostic indicators of prostate cancer and the expression of telomerase appear to be linked. The presence of telomerase activity in prostate tissue may aid in the detection of prostate cancer and produce additional prognostic information.     

Detection of telomerase activity in prostatic fluid specimens

We report here the extended use of the telomeric repeat amplification protocol (TRAP) assay for the detection of telomerase activity in fresh prostatic fluid obtained from anesthetized patients. Telomerase activity was detected in pellet extract and/or supernatant fluid of specimens obtained from 25 of 30 prostate cancer (PCa) patients (83%), whereas no activity was similarly detectable in specimens taken from 8 of 9 patients (89%) without clinical evidence of PCa. The positive predictive value (PPV) of the TRAP assay for PCa in this pilot study was 96%. We found a strong correlation between telomerase activity in prostatic fluid specimens and serum prostate specific antigen (PSA) values. Telomerase activity was found in 84% of specimens from patients with PSA values >4 ng/ml, whereas in specimens from patients with PSA values ⩽4 ng/ml, only 29% tested positive for telomerase activity. In patients with PSA values above the standard 0 to 4 ng/ml reference range, the PPV of the TRAP assay was 84%. Furthermore, telomerase activity could be detected in 85% of the specimens from PCa patients with Gleason scores >4. In prostatic fluid from PCa patients with Gleason scores of ⩽4, telomerase activity was detected 50% of the time. The PPV was 92% when a Gleason score of 4 was used as the lower limit. This pilot study raises the possibility of utilizing telomerase activity as an aid in PCa detection.     

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。     

The use of telomerase activity for the detection of prostatic cancer cells after prostatic massage

PURPOSE: Prostate cancer is the most commonly diagnosed cancer in the United States. The diagnosis or followup of prostate cancer in men older than 50 years is based on digital rectal examination, measurement of the free-to-total prostatic specific antigen ratio and transrectal ultrasound assisted needle biopsy of the prostate. We developed and evaluated a noninvasive method for diagnosing prostate cancer based on the measurement of telomerase activity after prostatic massage in fresh voided urine or after urethral washing. MATERIALS AND METHODS: We obtained 36 specimens of cells after prostatic massage in the fresh voided urine of 16 patients who subsequently underwent radical prostatectomy and after urethral washing in 20 who underwent prostate needle biopsies. Ethylenediaminetetraacetic acid was immediately added to the collected urine or washing to a final concentration of 20 mM. After protein extraction by CHAPS buffer each specimen was tested for telomerase activity in a 2-step modified telomeric repeat amplification protocol assay. The 2 prostate cancer cell lines PC-3 and LNCaP with high telomerase activity were used as a positive control. RESULTS: Telomerase activity was detected in 14 of 24 samples with known prostate cancer (sensitivity 58%). In contrast, no telomerase activity was found in the 12 cases without histological evidence of prostate tumor (specificity 100%). Eight of 9 poorly differentiated cancers expressed telomerase activity (89%), while only 6 of 15 well and moderately differentiated cancers showed telomerase activity (40%). CONCLUSIONS: Our data illustrate that telomerase activity may be detected in voided urine or washing after prostatic massage in patients with prostate cancer. Sensitivity was higher for poorly differentiated tumors. This approach is not currently available for detecting prostate cancer in clinical practice. However, these results are promising and further studies are ongoing.     

Telomerase activity in human renal cell carcinoma

PURPOSE: Telomerase activity has been detected in a wide variety of human tumors. The present study evaluated telomerase activity in association with the acquisition of renal cell carcinoma (RCC). METHODS: Telomerase activity was examined in 30 RCC and the adjacent normal kidney tissue, obtained as surgical specimens. The activity was assayed by polymerase chain reaction-based telomeric repeat amplification protocol assay. RESULTS: Among the 30 RCC, 18 (60%) displayed telomerase activity, whereas none of the normal tissue samples exhibited it. Subdivision of the tumors according to telomerase activity did not reveal any obvious difference in distribution with regard to tumor size, stage, histocytological subtype, or DNA-ploidy. However, a statistically significant relationship was found between the frequency of telomerase-positive activity and both serum immunosuppressive acidic protein level in the patient and tumor grade (P<0.05). There was no significant difference in the recurrent-free survival and the disease-specific survival between patients with positive telomerase activity and patients with negative activity. CONCLUSION: The present results indicate that telomerase activity might be related to the progression of RCC and thus a marker of malignant potential.     

端粒酶与前列腺癌及其生物学行为的关系

目的 探讨端粒酶（ＴＥ）与前列腺癌（ＰＣａ）及其生物学行为的关系。方法 应用端粒重复片段扩增法（ＴＲＡＰ）法,检测３９例ＰＣａ组织,１５例前列腺增生（ＢＰＨ）组织及１０例正常前列腺（ＮＰ）组织中的ＴＥ活性,并比较ＴＥ活性水平与ＰＣａ病理分化程度,临床分期及转移情况的关系。     

The expression of syndecan-1 and -2 is associated with Gleason score and epithelial-mesenchymal transition markers,E-cadherin and β-catenin,in prostate cancer

The epithelial-mesenchymal transition (EMT) is considered a key step in tumor progression, where the invasive cancer cells change from epithelial to mesenchymal phenotype. During this process, a decrease or loss in adhesion molecules expression and an increase in migration molecules expression are observed. The aim of this work was to determine the expression and cellular distribution of syndecan-1 and -2 (migration molecules) and E-cadherin and β-catenin (adhesion molecules) in different stages of prostate cancer progression. A quantitative immunohistochemical study of these molecules was carried out in tissue samples from benign prostatic hyperplasia and prostate carcinoma, with low and high Gleason score, obtained from biopsies archives of the Clinic Hospital of the University of Chile and Dipreca Hospital. Polyclonal specific antibodies and amplification system of estreptavidin-biotin peroxidase and diaminobenzidine were used. Syndecan-1 was uniformly expressed in basolateral membranes of normal epithelium, changing to a granular cytoplasmatic expression pattern in carcinomas. Syndecan-2 was observed mainly in a cytoplasmatic granular pattern, with high immunostaining intensity in areas of low Gleason score. E-cadherin was detected in basolateral membrane of normal epithelia showing decreased expression in high Gleason score samples. β-Catenin was found in cell membranes of normal epithelia changing its distribution toward the nucleus and cytoplasm in carcinoma samples. We concluded that changes in expression and cell distribution of E-cadherin and β-catenin correlated with the progression degree of prostate adenocarcinoma, suggesting a role of these molecules as markers of progression and prognosis. Furthermore, changes in the pattern expression of syndecan-1 and -2 indicate that both molecules may be involved in the EMT and tumor progression of prostate cancer.     

Detection of telomerase activity in esophageal squamous cell carcinoma and normal esophageal epithelium

Background/aims: Relatively little has been reported about the telomerase activity of esophageal squamous cell carcinoma or normal esophageal epithelium. In this study, we have evaluated whether telomerase activity is a useful marker for detecting malignancies in the esophagus. Patients/methods: Esophageal carcinomas and normal esophageal tissues adjacent to carcinomas were obtained from 52 surgically treated, unselected patients, and normal esophageal epithelium from 11 non-cancerous patients were obtained by means of biopsy. The telomeric repeat amplification protocol (TRAP) assay was used for detection of telomerase activity in these samples. The incidence of detection of telomerase activity in esophageal carcinoma was compared with that of telomerase activity in normal esophageal epithelium. Moreover, the clinicopathological characteristics of telomerase-positive tumors were compared with those of telomerase-negative tumors. Results: Of the 52 carcinomas, 40 (77%) had detectable telomerase activity. However, telomerase activity was detected in 45 of 52 (87%) normal tissue samples adjacent to carcinomas and in 8 of 11 (73%) normal esophageal epithelium from non-cancerous patients. In esophageal cancer, no significant difference was detected in the clinicopathological findings between the telomerase-positive and -negative cases. Conclusion: These results indicate that not only esophageal squamous cell carcinomas but also normal esophageal epithelium show strong telomerase activity. Thus, telomerase activity may not be a good marker for the detection of carcinoma in the esophagus. Received: 16 March 1999 Accepted: 8 July 1999     

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.     

Telomerase activity in malignant and benign renal tumors

OBJECTIVES: An important characteristic of malignant cells is their unlimited replicative potential, their immortality. In conferring this immortality, the enzyme telomerase is believed to play a crucial role. The detection of telomerase activity provides new knowledge regarding the biologic growth behavior of tumors and offers new diagnostic and therapeutic tools. METHODS: In the present study the sensitive TRAP assay (telomeric repeat amplification protocol) was used to examine 44 malignant renal tumors and 8 benign tumors of the kidney and 52 specimens of normal renal tissue for telomerase activity. RESULTS: Telomerase activity was detected in 63% of tissue samples obtained from histologically confirmed renal cell carcinomas. In cases of renal cell carcinoma restricted to the kidney, telomerase activity was detected in 58%. In cases in which tumor growth has progressed beyond the limits of the organ, telomerase activity was found in 69%. This stage dependence, however, did not reach statistical significance. No correlation to tumor grading was observed. Telomerase activity was found less frequent in chromophobe renal cell carcinomas. Neither the 8 benign renal tumors (4 oncocytomas and 4 angiomyolipomas) nor the specimens of normal kidney showed any evidence of telomerase activity. CONCLUSIONS: The proportion of remarkable slow-growing renal cell carcinomas showing telomerase activity is less than in other malignancies and may correlate with biologic growth behavior. Possible explanations include the presence of an alternative pathway, called ALT (alternative lengthening of telomeres) and an association with the loss or presence of the telomerase suppressor on the short arm of chromosome 3. Prolonged follow-up will be of special interest to determine whether lack of telomerase activity predicts favorable outcome.     

Telomerase activity is correlated with lower grade and lower stage bladder carcinomas

BACKGROUND: Telomerase is a ribonucleoprotein enzyme that compensates for the progressive erosion of telomeres. The increasing interest in telomerase is motivated by the demonstration that most human carcinomas are telomerase positive. The potential use of telomerase activity in bladder carcinomas using a urine sample has been reported in several studies. However, little is known about the detection of telomerase activity in bladder carcinoma tissues. Herein, we investigate telomerase activity in bladder carcinoma tissues according to grade (G) and stage. MATERIAL AND METHODS: Telomerase activity was assayed by polymerase chain reaction enzyme-linked immunosorbent assay methods. Malignant lesions were assessed in 37 patients with bladder carcinoma and no malignant lesions were assessed in five patients with dysplasia or inflammatory bladder lesions. RESULTS: Twenty-three out of 37 carcinoma samples were telomerase-positive and one out of five control samples without carcinoma was telomerase-positive. The positive rates according to stage and grade were 83.3% for superficial and 42.1% for invasive stages and 83.3% for G1, 66.7% for G2 and 40.0% for G3. Telomerase activity was correlated with lower grade and lower stage bladder carcinomas. CONCLUSION: These results strongly suggest that reactivation of telomerase may differ between superficial and invasive bladder carcinomas and also between low grade and high grade bladder carcinomas.     

The clinical implications of telomerase activity in upper tract urothelial cancer and washings

OBJECTIVE: To measure telomerase activity in upper tract urothelial carcinomas (as renal pelvic tumours comprise nearly half of all kidney tumours in Taiwan, a much higher percentage than in other countries) and to determine whether telomerase activity could be used as an additional diagnostic marker in exfoliated cancer cells present in upper tract urothelial washing fluids, thus providing earlier diagnosis and treatment. Materials and methods Telomerase activity was assessed using the telomeric repeat amplification protocol assay in tissue samples from 31 upper tract urothelial carcinomas (from 29 patients). The feasibility of identifying cancer using telomerase activity in exfoliated cancer cells in 17 upper tract urothelial washing samples was also investigated. RESULTS: Telomerase activity was found in 30 (97%) of the 31 upper tract urothelial cancer tissue samples; telomerase activity was detectable in 95% of superficial cancers and in all 11 invasive tumours. The sensitivity of measuring telomerase activity was 100% for grade 1, 93% for grade 2 and 100% for grade 3 tumours. In contrast, telomerase activity was detected in only two (8%) of 26 normal adjacent tissue samples. When the telomerase activity of urothelial washing fluid was compared with that in the corresponding tumours, there was compatible telomerase activity in 15 of the 17 samples. Telomerase activity was more sensitive than voided urine cytology (15%) and washing fluid cytology (53%). In addition, the telomerase activity was high in metastatic lesions. CONCLUSION: Telomerase activity is present in most upper tract urothelial cancer tissues and may be present at an early stage of carcinogenesis. Telomerase activity can be detected in exfoliated cells in urothelial washing fluids in a high proportion of patients with upper tract urothelial cancer. These results suggest that measuring telomerase activity in the exfoliated cancer cells obtained from urothelial washing could be a potentially useful addition to the conventional diagnostic tools used to identify patients with upper tract urothelial carcinoma.     

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     

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。     

Tertiary Gleason pattern 5 is a powerful predictor of biochemical relapse in patients with Gleason score 7 prostatic adenocarcinoma

PURPOSE: In radical prostatectomy specimens Gleason score 7 is among the most commonly assigned scores for prostate carcinoma accounting for 30% to 50% of cases. Gleason score 7 is different from other more differentiated prostate carcinomas (tumors of Gleason scores 5 and 6) with a significantly worse outcome and higher rate of recurrence. Nonetheless, Gleason score 7 tumors are heterogeneous. In this study we examined the differences in clinical outcome between primary Gleason grade 3 and 4 tumors in patients who underwent radical prostatectomy, and determined the influence of tertiary Gleason pattern 5 on patient outcome. MATERIALS AND METHODS: A total of 504 patients underwent radical prostatectomy for prostate cancer and 228 of the patients (45%) had a Gleason score of 7. Cases were analyzed for a variety of clinical and pathological parameters. The influence of primary Gleason pattern and tertiary Gleason pattern 5 on patient outcome was assessed in the Cox regression model. RESULTS: Among 228 patients with Gleason score 7 prostatic adenocarcinoma, 91 (40%) had a primary Gleason pattern 4 and 137 (60%) had primary Gleason pattern 3. Patients of the former group were more likely to have a higher pathological stage (p = 0.003), more likely to have PSA recurrence (p = 0.02) and more likely to have a tertiary Gleason pattern 5 (p <0.0001). A total of 37 (41%) patients with primary Gleason 4 had a tertiary Gleason pattern 5, whereas only 13 (9%) patients with primary Gleason 3 had a tertiary Gleason pattern 5. In the Cox regression model controlling for tumor stage and surgical margin status, the primary Gleason pattern was not an independent predictor of PSA recurrence (p = 0.80), whereas the presence of tertiary Gleason pattern 5 was a significant predictor of PSA recurrence (hazard ratio 2.10, 95% CI 1.24-3.55, p = 0.006). Five-year PSA recurrence-free survival was 70% for patients without a tertiary Gleason pattern 5 compared to 19% for those patients with a tertiary Gleason pattern 5. CONCLUSIONS: Among patients with Gleason score 7, primary Gleason grade 4 indicates a likelihood of higher tumor stage and higher probability of PSA recurrence than does primary pattern 3. However, it does not independently predict a worse outcome after controlling for other known prognostic parameters associated with disease progression. Regardless of whether the primary Gleason pattern is 3 or 4, a tertiary Gleason pattern 5 is the strongest predictor of a worse outcome in patients with Gleason grade 7 prostatic adenocarcinoma. Therefore, tertiary pattern 5 should be reported in radical prostatectomy specimens.