大肠癌与端粒酶活性相关性的研究

目的　探讨端粒酶活性在大肠癌发生、发展以及浸润转移中的意义。方法　应用端粒重复扩增 (TRAP)及免疫组织化学法对 30例大肠癌、癌旁组织、正常大肠组织及 2 0例大肠腺瘤性息肉组织端粒酶活性、端粒酶催化亚基蛋白 (hTERT)的表达进行检测。结果　端粒酶活性在癌组织中检出率明显高于其他组织 (P <0 .0 5) ;大肠癌组织端粒酶活性与淋巴结是否有转移之间关系密切 ,伴淋巴结转移大肠癌端粒酶阳性表达率明显高于无淋巴结转移者 (P <0 .0 5)。结论　端粒酶活性与大肠癌的发生发展以及浸润转移密切相关 ,检测端粒酶活性对临床预测大肠癌淋巴结转移趋势、评价恶性程度和判断预后均有重要意义     

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     

端粒酶活性表达在肾恶性肿瘤中的临床意义

目的　探讨端粒酶活性表达在肾恶性肿瘤中的临床意义。　方法　采用端粒酶 PCR ELISA法检测 3 1例肾恶性肿瘤、3 1例肿瘤旁组织和 6例正常肾组织标本端粒酶活性 ,按不同的临床病理参数分组分析。　结果　 3 1例肾恶性肿瘤组织端粒酶表达阳性率 80 .6% ,肿瘤旁组织及正常肾组织端粒酶表达均阴性 ,差异有显著性意义 (P <0 .0 1) ;病理分级Ⅰ级者端粒酶表达阳性率 58.3 %(7/ 12 ) ,Ⅱ级者阳性率 91.7% (11/ 12 ) ,Ⅲ～Ⅳ级者阳性率 10 0 .0 % (7/ 7) ,Ⅱ～Ⅳ级者端粒酶表达明显高于Ⅰ级者 (P <0 .0 5) ;RobsonⅠ期阳性率 77.8% (14 / 18) ,Ⅱ期阳性率 81.8% (9/ 11) ,Ⅲ～Ⅳ期 2例均阳性 ;T1N0 M0 期 3例均阳性 ,T2 N0 M0 期阳性率 79.2 % (19/ 2 4) ,T3N0 M0 期阳性率 66.7% (2 / 3 ) ,T2 N2 M0期 1例阳性 ,端粒酶表达与肿瘤临床分期无明显相关 (P >0 .0 5)。　结论　端粒酶活性检测结合病理检查对肾恶性肿瘤的早期诊断及预后判断有重要价值。     

膀胱癌组织中端粒酶活性研究

应用聚合酶链反应（ＰＣＲ）为基础的端粒酶活性检测方法（ＴＲＡＰ法），对５例正常膀胱组织、４例膀胱炎症组织、４２例膀胱癌组织和癌旁正常组织进行检测。正常膀胱组织和膀胱炎症组织无端粒酶活性，膀胱癌组织端粒酶表达阳性率７６．２％，癌旁组织阳性率为２６．２％，端粒酶活化与膀胱癌分期分级呈正相关。结果提示：端粒酶活化与膀胱癌浸润发展密切相关，癌旁组织端粒酶活化提示肿瘤微浸润可能。     

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     

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     

胃癌组织和外周血中端粒酶活性的研究

目的测定端粒酶在胃癌患者的肿瘤组织、外周血中的活性，以探讨端粒酶活性的表达及其在临床应用上的意义。方法应用TRAP—PCR—ELISA方法检测36例胃癌患者的癌组织及其相应癌旁组织、正常组织、外周血单核细胞中端粒酶活性以及20例正常对照者外周血单核细胞端粒酶的活性。结果癌组织、癌旁组织、正常组织中端粒酶阳性率分别为80．6％（29／36）、16．7％（6／36）和5．6％（2／36）。6例胃癌患者外周血有18例测到端粒酶阳性，阳性率为50％，正常人外周血未测到端粒酶阳性。癌组织与癌旁组织及正常组织之间、胃癌患者外周血与与正常人外周血之间的端粒酶阳性率差异有统计学意义（P〈0．01）。端粒酶活性与患者的性别、年龄、肿瘤大小、浸润深度、分化程度、淋巴转移和TNM分期等临床病理参数差异无统计学意义（P〈0．05）。结论检测患者胃癌组织、外周血中端粒酶活性有助于胃癌的早期诊断，对预后监控及治疗也有重要意义。     

肝细胞癌端粒酶活性与细胞周期抑制蛋白p27~(kip1)的相关性研究

目的　探讨端粒酶活性 (TA )和细胞周期蛋白依赖激酶抑制蛋白 p2 7kip1在肝癌中的表达及其相互关系。方法　采用端粒重复序列扩增 酶联免疫吸附 (TRAP ELISA)方法测定 2 7例肝细胞癌 (HCC)和 2 3例肝硬化组织中的端粒酶活性 ,采用免疫组织化学方法测定 p2 7kip1在HCC中的表达。结果　 2 7例HCC中 2 4例端粒酶阳性 ,3例阴性 ;2 3例肝硬化中 3例阳性 ,2 0例阴性 ,两者差异有非常显著性 ( r =0 .484,P <0 .0 0 1) ;HCC中平均 p2 7kip1标记指数为 5 0 .30± 19.83;端粒酶活性与p2 7kip1呈正相关 (P <0 .0 5 )。结论　测定端粒酶活性有利于早期发现肝癌 ,端粒酶的激活与p2 7kip1蛋白表达有关。     

乳腺癌端粒长度与端粒酶活性变化的相关性研究

目的　检测乳腺癌及其癌旁组织中的端粒 (TLM )长度、端粒酶 (TLMA )活性的表达 ,探讨乳腺癌端粒长度与端粒酶活性的关系。方法　采用端粒酶重复扩增实验 (TRAP) 银染法检测端粒酶活性 ,以地高辛标记的Southern杂交的方法检测端粒长度。结果　端粒酶活性在乳腺癌TNM分期的进展中由Ⅰ期的 5 9.1%增高到Ⅳ期的 92 .9% ,而端粒长度在乳腺癌TNM分期的进展中不断缩短 ,由Ⅰ期的 (7.5 4± 0 .95 )kb缩短到Ⅳ期的 (5 .0 3± 0 .5 3 )kb。恶性乳腺肿瘤癌旁组织中的平均端粒长度皆位于正常水平。乳腺癌中端粒酶阳性表达组的端粒长度为 (4 .45± 1.3 9)kb显著短于端粒酶阴性组的 (5 .70± 1.2 3 )kb。结论　恶性乳腺肿瘤将端粒酶激活并没有绝对延长其染色体末端的端粒长度 ,而且 ,随着肿瘤的发展端粒片段还会缩短 ,并与端粒酶活性似乎存在着反向的关系。     

肝细胞癌中nm-23H1的表达及临床意义

利用免疫组织化学 (免疫组化 )法检测肝细胞癌组织中nm 2 3H1 的表达情况 ,探讨其临床意义。结果显示 :89例肝细胞癌组织中nm 2 3H1 阳性表达 3 4例 (3 4/ 89,3 8.2 0 % ) ,阴性表达 5 5例(5 5 / 89,61.80 % ) ;有直接浸润或远处转移的 49例病例中 12例nm 2 3H1 表达阳性 (12 / 49,2 4.49% ) ,无直接浸润或转移的 40例病人中 2 2例表达阳性 (2 2 / 40 ,5 5 .0 0 % ) ;5 1例能手术切除的肝癌中 2 8例nm 2 3H1 表达阳性 (5 4.90 % ) ,3 8例未能手术切除的病例中 6例表达阳性 (15 .79% )。提示有转移浸润的肝细胞癌组织nm 2 3H1 基因阳性表达远低于无浸润转移者 ,说明nm 2 3H1 基因与肝细胞癌的转移浸润有着一定的关系 ;病灶能手术切除者其nm 2 3H1 基因阳性表达率显著地高于不能手术切除者 (P <0 .0 1) ,因此认为肝细胞癌的nm 2 3H1 表达可作为术前判断肝癌能否切除的参考指标。     

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 in urological malignancy.

PURPOSE: Telomerase is a ribonucleoprotein enzyme that compensates for the progressive erosion of chromosomal ends, called telomeres. In most somatic cells telomerase expression is repressed and telomeres progressively shorten after each cell division, causing cell senescence. Conversely telomerase is active in most human cancers, maintaining the integrity of chromosome ends and representing an important step in cell immortalization and carcinogenesis. The large and increasing interest in telomerase was motivated by the demonstration that more than 90% of human cancers are telomerase positive, whereas most normal tissues or benign tumors contained low or undetectable telomerase activity. We addressed the most recent data on telomerase detection in urological malignancy. Approaches to telomerase inhibition as a future anti-cancer therapy are also discussed. MATERIALS AND METHODS: We comprehensively reviewed the most recent and significant publications in this field using current issues of specific journals and a MEDLINE search. RESULTS: Telomerase is often expressed in bladder (90%), prostate (80%) and renal (69%) carcinoma. A variable but significant percent of normal tissues from tumor adjacent zones or noncancer samples are positive for telomerase. The clinical role of telomerase is still questionable in renal cancer, while important insights into the diagnostic role of telomerase in bladder and prostate carcinoma are increasing. Telomerase detection in exfoliated cells collected with urine or bladder washings seems a promising tool for the diagnosis and management of bladder cancer. CONCLUSIONS: Larger perspective studies of larger groups of patients are required to discover an appropriate role for telomerase when assessing these tumors. The improvement of quantitative methods to evaluate the expression of telomerase is a cornerstone in the complete clarification of the clinical relevance of telomerase.     

PCR-ELISA法检测膀胱肿瘤患者尿脱落细胞端粒酶活性

目的：探讨尿脱落细胞端粒酶活性变化在膀胱肿瘤诊断中的作用。方法应用PCR－ELISA法检测53例膀胱肿瘤患者尿液脱落细胞端粒酶的活性。结果：非膀胱肿瘤和膀胱肿瘤患者尿液脱落细胞端粒酶活性阳性率分别为64．15％（34．53）和7．69％（2／26）,健康对照者7例均为阴性,膀胱肿瘤患者与正常人及非膀胱肿瘤患者的端粒酶活性分别相比,差别均有极显著性意义（P＜0．001）。但端粒酶活性与肿瘤的分期分级无相关性。结论：尿脱落细胞端粒酶活性检测可以作为诊断膀胱肿瘤的无创性检测方法,但不能预测膀胱肿瘤的临床分期分级。     

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。     

Telomerase activity Simplification of assay and detection in bladder tumor and urinary exfoliated cells

Detection of telomerase activity can differentiate malignant from benign cells. However, the original telomeric repeat amplification protocol (TRAP) methods had a number of limitations including a radioisotope labeling [α(32)P] dCTP [α(32)P] dGTP system. We developed digoxigenin labeled CX primer to detect telomerase activity without using radioisotope and attempted to detect telomerase activity of bladder tumor and exfoliated cells in bladder cancer patients. Telomerase activity was detected in 5 (71%) of 7 patients diagnosed with grade 1, 31 (97%) of 32 grade 2, and 11 (100%) of 11 grade 3 bladder tumors. In urinary exfoliated cells, 32 (82%) of 39 grades 1 or 2 bladder tumors were positive for telomerase activity but 20 (51%) of 39 were positive for urinary cytology (P < 0.01). Ten (91%) of 11 of grade 3 tumors were positive for telomerase activity and 11 (100%) of 11 were positive urinary cytology. Three of 100 noncancerous patients were positive for telomerase activity. Sensitivity, specificity, and positive predictive value of telomerase activity assay in urinary exfoliated cells were 84%, 97%, and 93%, respectively. Telomerase activity may be a useful diagnostic marker to detect the existence of immortal cancer cells in the urine.     

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.     

The molecular detection of circulating tumor cells in bladder cancer using telomerase activity.

PURPOSE: The detection of circulating tumor cells and micrometastases may have important prognostic and therapeutic implications. We investigated telomerase activity as a molecular marker for detecting bladder carcinoma cells in blood. MATERIALS AND METHODS: Peripheral blood mononuclear cells were isolated from whole blood using Ficoll/Hypaque. Immuno-magnetic beads labeled with an epithelial specific antibody were used to harvest epithelial cells from peripheral blood mononuclear cells. Telomerase activity was detected in this select population using the telomerase-polymerase chain reaction-enzyme-linked immunosorbent assay test based on the telomerase repeat amplification protocol method. The clinical applicability of this technique was explored by evaluating 30 patients with muscle invasive or metastatic bladder carcinoma and 17 healthy volunteers. RESULTS: Telomerase expression was detected in 27 of the 30 patients (90%) with high grade, muscle invasive or metastatic bladder cancer but in none of the 17 healthy controls. CONCLUSIONS: This test is a minimally invasive and specific approach for detecting circulating epithelial cells in patients with bladder cancer. This method may have great value for monitoring cancer progression.     

FOCAL INTRATUMORAL HETEROGENEITY FOR TELOMERASE ACTIVITY IN HUMAN PROSTATE CANCER

PURPOSE: The value of telomerase activity as a marker in clinical decision-making is closely related to how representative the analysis of a small tumor sample is for the whole tumor. We therefore evaluated the intratumoral distribution pattern of telomerase activity in prostatic carcinomas. MATERIALS AND METHODS: From 50 prostate cancer patients treated with radical prostatectomy, telomerase activity was determined using the telomeric repeat amplification protocol (TRAP assay). Comparative analysis of at least two separate cancer areas from a single tumor was performed in 42 cases. RESULTS: Telomerase activation has been demonstrated in 90% of the prostatic carcinomas. Focal intratumoral heterogeneity was found in 38.1% of the tumors with at least two different areas examined. Telomerase positivity of all samples from one given tumor was detected in 50%, telomerase negativity of all samples in 11.9%. A heterogeneous telomerase activity pattern was more frequently detected in tumors with a Gleason score < or = 7 than in those with a Gleason score > 7. Furthermore, there was an increase in the proportion of homogeneously telomerase-positive tumors with increase in severity of the Gleason score. The differences reached statistical significance. Telomerase activity was also detected in non-cancerous prostatic tissue samples. CONCLUSIONS: Telomerase activation is nearly ubiquitous in prostatic carcinomas, although a heterogeneous telomerase activity pattern within tumors might produce a false-negative result in the telomerase activity assay. This limits the value of telomerase activity assays for diagnostic means. There is evidence for a shift from telomerase-negative prostate cancer tissue toward telomerase positivity during the progression process of prostate cancer. The relatively high proportion of telomerase-positive nonmalignant prostatic tissue samples argues against cancer-specificity of telomerase activation.