PCR-ELISA检测尿液中脱落细胞端粒酶活性及临床意义

为探讨尿液中脱落细胞端粒酶活性检测对膀胱癌的诊断与术后跟踪随访的意义,以PCR-ELISA方法检测膀胱癌组织和自然排尿中膀胱癌脱落细胞端粒酶活性,以巴氏染色法对脱落细胞进行细胞学检查.62例膀胱癌患者中,有59例肿瘤组织表达端粒酶活性(95.2%);47例尿脱落细胞表达端粒酶活性(75.8%),24例尿脱落细胞中发现癌细胞(38.7%),脱落细胞的端粒酶活性检测明显高于细胞学检查(P＜0.005),随着肿瘤分化程度和临床病理分期的增加,阳性率明显增高(P＜0.05).结果表明自然排尿端粒酶活性的检测,操作简单、灵敏、特异性强,可望成为诊断膀胱癌的首选方法.     

膀胱癌患者尿液脱落细胞端粒酶活性测定

端粒酶主要由蛋白质和RNA组成,是一种反转录酶,能以其自身所携带的RNA为模板,不断合成新的端粒DNA序列添加到染色体末端,阻止端粒的缩短,使细胞成为永生性细胞或癌细胞[1].近年来,大量科学研究证实端粒酶活性表达与肿瘤发生密切相关,它在恶性肿瘤组织中及相关脱落细胞中检出率很高,可作为肿瘤早期诊断的生物学指标[2,3].据此,我们对58例膀胱移行上皮细胞癌患者尿液中脱落细胞端粒酶活性进行了测定,以探讨一种敏感、特异、非侵入性的诊断和随访膀胱肿瘤的方法.     

膀胱癌患者尿液脱落细胞端粒酶活性测定

端粒酶主要由蛋白质和RNA组成,是一种反转录酶,能以其自身所携带的RNA为模板,不断合成新的端粒DNA序列添加到染色体末端,阻止端粒的缩短,使细胞成为永生性细胞或癌细胞[1]。近年来,大量科学研究证实端粒酶活性表达与肿瘤发生密切相关,它在恶性肿瘤组织中及相关脱落细胞中检出率很高,可作为肿瘤早期诊断的生物学指标[2,3]。据此,我们对58例膀胱移行上皮细胞癌患者尿液中脱落细胞端粒酶活性进行了测定,以探讨一种敏感、特异、非侵入性的诊断和随访膀胱肿瘤的方法。材料和方法一、检测对象　肿瘤组为本院经确诊的膀胱移行细胞癌患者58例,男…     

膀胱癌患者尿脱落细胞中survivin基因的检测

目的 通过对膀胱癌患者尿液脱落细胞中人体细胞凋亡相关基因survivin的检测，探讨可早期发现、常规筛检膀胱癌，且不给患者带来痛苦的方法。方法 留取31例膀胱癌患者、20例泌尿系其他良性疾病患者和10例健康志愿者的新鲜中段尿液200ml ,用逆转录聚合酶链反应（RT-PCR）检测survivin的表达；同时行尿脱落细胞学检查。结果 用RT-PCR检测尿液中survivin的敏感性为100%，与尿脱落细胞学检相比较，具有很高的敏感性。结论 检测尿液中survivin可用于膀胱癌的术前诊断。     

应用尿脱落细胞端粒酶活性的检测诊断膀胱癌

目的 寻找一种敏感特异的非介入性方法诊断膀胱癌，探讨检测自排尿脱落细胞端粒酶活性在膀胱癌早期诊断及术后随访中的意义。方法 应用聚合酶链反应。酶联免疫吸附试验(PCR—ELISA)，分别对膀胱癌患者、良性血尿患者及健康志愿者的自排尿脱落细胞进行端粒酶活性的检测。结果 18例膀胱癌患者的尿脱落细胞端粒酶活性阳性为14例(14／18)，20例良性血尿患者及20名健康志愿者均无端粒酶活性表达。结论 尿脱落细胞端粒酶活性的检测敏感性高，特异性好，可用于膀胱癌的早期诊断及术后随访。     

乳头溢液脱落细胞端粒酶活性检测

目的探讨乳头溢液脱落细胞端粒酶活性检测在临床诊断中的应用。方法采用ＴＲＡＰ－ＰＣＲ银染定性法及ＴＲＡＰ－ＰＣＲ－ＥＬＩＳＡ定量法对５０例溢乳进行脱落细胞端粒酶活性分析。结果５０例溢乳脱落细胞端粒酶阳性率高达７８％（３９／５０）,随着年龄的增大,端粒酶阳性率呈逐步下降趋势,而同期送检的细胞病理诊断则无１例找到癌细胞。结论溢乳脱落细胞端粒酶阳性检出率明显增高。随着疾病的发展,端粒酶阳性患者易发生恶变的可能性是否大于端粒酶阴性患者需进一步研究。     

膀胱癌尿脱落细胞端粒酶活性检测及其意义

目的　研究端粒酶对膀胱癌的诊断价值。方法　应用端粒重复扩增 (TRAP) 微孔板杂交法测定 58例膀胱癌患者肿瘤组织及尿液、膀胱冲洗液脱落细胞的端粒酶活性 ,并与细胞学检查作比较。结果　肿瘤组织标本中 ,端粒酶阳性率为81 0 3 % ;尿液标本中 ,阳性率为 57 14 % ;膀胱冲洗液标本中 ,阳性率为 70 3 7%。同期尿细胞学检查 ,阳性率仅 3 6 2 1%。在低级别肿瘤中 ,尿或膀胱冲洗液端粒酶阳性率明显高于细胞学检查。结论　膀胱癌组织有端粒酶活性的高表达 ,而尿液、膀胱冲洗液脱落细胞中有较高的端粒酶活性检出率。是膀胱肿瘤的一种新的无创伤性的诊断方法     

PCR—ELISA检测体液脱落细胞端粒酶活性及临床意义

端粒酶是近年来发现的与肿瘤发生发展密切相关的一种逆转录酶,已成为肿瘤研究的热点。目前国内外对实体瘤的端粒酶活性进行了广泛的研究,但对体液脱落细胞中端粒酶的活性的研究尚较少。我们应用ＰＣＲＥＬＩＳＡ方法检测体液脱落细胞的端粒酶活性,旨在探讨该方法在肿瘤的早期诊     

PCR-ELISA检测尿液中膀胱癌脱落细胞端粒酶活性及临床意义

端粒酶是近年来发现的与肿瘤发生关系密切相关的一种逆转录酶,目前国内外对实体瘤的端粒酶活性进行了广泛的研究,但对体液脱落细胞中端粒酶活性的研究较少。我们应用PCR-ELISA方法检测尿液中脱落细胞的端粒酶活性,旨在探讨该方法在膀胱癌早期诊断和随访中的应用价值。     

尿脱落细胞DNA流式细胞术分析对膀胱癌诊断的临床意义

目的　探讨尿脱落细胞DNA分析在膀胱癌诊断中的临床意义。方法　利用流式细胞仪对良性血尿、膀胱肿瘤及膀胱癌患者尿脱落细胞进行DNA倍体和SPF分析。结果　良性血尿患者为正常二倍体DNA分析图谱。膀胱肿瘤患者二倍体肿瘤 2 2例 ,异倍体肿瘤 4例 ;而膀胱癌患者二倍体肿瘤 8例 ,异倍体肿瘤 38例 ;膀胱癌、膀胱肿瘤患者的SPF(15 5 3± 6 5 4 ,9 37± 5 0 5 )明显高于良性血尿患者 (5 6 7± 2 4 8,P <0 0 0 1,P <0 0 0 1) ,而膀胱癌患者的SPF明显高于膀胱肿瘤 (P <0 0 0 1)。结论　尿脱落细胞DNA分析可望成为诊断膀胱癌的一种辅助方法     

Bioluminescent method for detecting telomerase activity

BACKGROUND: Telomerase is a promising biomarker in cancer diagnosis and therapy. The elongation of telomeric repeats catalyzed by telomerase is accompanied by release of six PP(i) for each TTAGGG repeat (1 pmol PP(i)/310 pg telomeric repeats). We developed a novel method to measure telomerase activity by use of an enzymatic luminometric PP(i) assay (ELIPA). METHODS: Extracts of cell lines and tissues were incubated with primer at 30 degrees C for 30 min. Released PP(i) was converted to ATP by sulfurylase, and ATP was detected by a luciferase bioluminescence system. The ELIPA results were compared with results obtained with the conventional telomeric repeat amplification (TRAP)-ELISA in 42 lung carcinoma tissues and 27 control tissues without malignancy. RESULTS: The lower detection limits of ELIPA and TRAP-ELISA were 5 and 10 cells, respectively. The within-run imprecision (CV) of ELIPA was < or =12%. When compared with TRAP-ELISA, the correlation coefficient (r) was 0.79. When we used the cutoff value from ROC analysis to distinguish malignant and nonmalignant tissues, the sensitivity and specificity of ELIPA were 83% and 96%, respectively, whereas the sensitivity and specificity of TRAP-ELISA were 71% and 96%, respectively. CONCLUSION: ELIPA is a simple and sensitive homogeneous method to quantify telomerase activity.     

膀胱移性细胞癌尿脱落细胞端粒酶催化亚基检测及临床意义

目的　检测尿脱落细胞端粒酶催化亚基 (hTERT)并探讨其临床意义。方法　用RT PCR法分别对膀胱癌组织、正常膀胱组织以及膀胱移性细胞癌和泌尿系良性疾病患者尿脱落细胞进行hTERT检测。结果　 2 7例膀胱癌组织hTERT表达呈10 0 %阳性 ,正常膀胱粘膜 16例均无阳性表达。 3 2例膀胱移行性细胞癌 (transitionalcellcarcinomaofbladder,TCC)患者尿脱落细胞hTERT表达阳性率为 87 5 % ( 2 8/ 3 2 ) ,非肿瘤组表达阳性率为 13 3 % ( 2 / 15 ) ,与TCC病理分级、分期无相关性。结论　检测脱落细胞hTERT是一种无创伤性的、敏感度高、特异性强的方法 ,是TCC早期诊断的一个有价值的辅助指标。     

Allelic deletion fingerprinting of urine cell sediments in bladder cancer.

BACKGROUND: Bladder cancer shows frequent nonrandom allelic deletion at various chromosomal regions. Genotypic detection methods could potentially identify patients at risk for recurrent progressive disease. In this study, we examined allelic deletion at specific chromosomal loci in tumor tissue and urine cell sediment samples using a microsatellite-based protocol. Although both allelic deletion and microsatellite instability have been reported in primary bladder cancer, microsatellite instability was not specifically examined in this study. We report a pilot study of 40 patients with bladder cancer in which allelic deletion in tumor tissue and urine cell sediment was compared with conventional urine cytology results. METHODS AND RESULTS: Forty tumors were analyzed using a set of microsatellite primers from chromosomes 3, 4, 8, 11, 14, and 17 to construct allelic deletion fingerprints. Cy5.5-labeled PCR products were analyzed using the OpenGene System and GeneObjects software. Eighty-eight percent of tumors showed allelic deletion. In urine cell sediments, the tumor detection rate was 80% compared with 50% for routine urine cytology. The allelic deletion fingerprinting (ADF) procedure identified 69% of incipient tumors, cases initially classified as normal by routine urine cytology. CONCLUSION: ADF analysis provides a reliable noninvasive method for the detection and monitoring of recurrent cancer in urine cell sediment samples from patients with bladder cancer.     

尿液端粒酶催化亚基mRNA检测与膀胱肿瘤的诊断

膀胱肿瘤是泌尿系统最常见的恶性肿瘤之一 ,近年来发病率有增高的趋势 ,早期诊断是提高治疗效果的关键 ,另外膀胱肿瘤复发率高 ,10 %～ 2 0 %的复发肿瘤会发展到更高级和肌层浸润。国内外大量研究文献证实 ,端粒酶的激活与肿瘤的发生、发展有关 ;而端粒酶催化亚基 (hTERT)在调节端粒酶活性中起决定作用。现就尿液hTERT及其在膀胱肿瘤的早期诊断、复发监测和预后判断等方面的价值作一综述。     

Label-free highly sensitive detection of telomerase activity in cancer cell by chemiluminescence imaging

We have developed a new methodology for label-free highly sensitive telomerase activity assay using chemiluminescence imaging. This method can detect the telomerase activity from as little as 10 cultured cancer cells without PCR. Furthermore, telomerase inhibition is shown, demonstrating the potential for screening of telomerase inhibitors as anticancer drug agents.     

放射性核素示踪技术检测端粒酶活性的研究进展

人端粒酶(telomerase)与人类的衰老与肿瘤发生密切联系;其主要组成成分之一——人端粒酶催化亚单位(hTERT)是端粒酶的限速亚单位,具有重要的临床应用价值与研究意义。检测端粒酶活性,尤其是hTERT活性,不仅有助于临床诊断肿瘤,而且对于进一步开展端粒酶研究具有重要作用。本文综述放射性核素示踪技术检测端粒酶活性的研究进展。