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目的 :组装简便实用的端粒酶活性相对定量检测试剂盒 ,建立了膀胱肿瘤端粒酶活性非同位素相对定量检测技术。方法 :用银染 -TRAP略行改良 ,对 54例膀胱癌组织、癌旁组织、12例正常膀胱黏膜 ,其中 4 6例膀胱癌患者同时收集了尿液及膀胱冲洗液的脱落细胞进行端粒酶活性相对定量值分析。结果 :54例膀胱癌组织中端粒酶活性相对定量值阳性率与正常膀胱黏膜和癌旁组织端粒酶活性相对定量值相比 ,差异有显著性 (P <0 0 0 1,P <0 0 0 1) ;癌旁组织略高于正常膀胱黏膜 ,但差异无显著性 (P >0 0 5) ,4 6例膀胱癌膀胱冲洗液与尿液相比差异有显著性 (P <0 0 1) ;4 6例膀胱癌尿液与 10例正常尿液相比差异有显著性 (P <0 0 1) ;54例膀胱癌与癌旁组织中端粒酶活性相对定量均值及标准差相比 ,差异有显著性 (P <0 0 0 1) ;4 6例膀胱癌冲洗液与尿液端粒酶活性相对定量均值及标准差相比差异有显著性 (P <0 0 5)。结论 :建立了膀胱肿瘤端粒酶活性的非同位素相对定量分析方法 ,具有特异性强、灵敏性高的特点 ,该方法可作为恶性肿瘤诊断和预后的指标之一。
OBJECTIVE: To establish a simple and practical kit for relative quantification of telomerase activity, and to establish a relative quantitative assay for non-isotope of telomerase activity in bladder tumor. Methods: Twenty-four cases of bladder cancer, including 54 cases of bladder cancer, paracancerous tissues and 12 cases of normal bladder mucosa, were treated with silver staining-TRAP. Twenty-six cases of bladder cancer were collected with exfoliated cells of urine and bladder irrigating solution for telomere Relative quantitative analysis of enzyme activity. Results: The positive rate of telomerase activity in 54 cases of bladder cancer was significantly higher than that of normal bladder mucosa and paracancerous tissues (P <0 01 1, P 0 There was no significant difference (P> 0.05) between the adjacent tissues and the normal bladder mucosa, and there were significant differences between the bladder irrigation solution and the urine in 46 cases of bladder cancer (P <0 0) 1). There were significant differences in the urine of 46 cases of bladder cancer and 10 cases of normal urine (P <0.01). The relative quantitative mean and standard deviation of telomerase activity in 54 cases of bladder cancer and paracancerous tissues (P <0.01). There was a significant difference (P <0.05) in the relative quantitative mean and standard deviation of the telomerase activity in 46 cases of bladder cancer and urine. Conclusion: A non-isotope quantitative analysis method of telomerase activity in bladder tumor was established. It has the characteristics of strong specificity and high sensitivity. It can be used as one of the indicators of malignant tumor diagnosis and prognosis.