胸/腹水端粒酶活性检测及诊断意义

Clinical significance of telomerase activity assay in diagnosis of pleural fluid and ascite

目的 本研究通过对胸 /腹水脱落细胞端粒酶活性的检测 ,探讨胸 /腹水脱落细胞端粒酶活性在胸 /腹水良恶性鉴别诊断中的临床价值。方法 收集 50例诊断明确的新鲜胸 /腹水标本 ,包括恶性胸水1 6例 (肺癌 8例、乳腺癌 5例、恶性淋巴瘤 3例 ) ,恶性腹水 2 4例 (原发性肝癌 3例、胃癌 8例、肠癌 9例、恶性淋巴瘤 4例 ) ,良性胸水 4例 (结核性胸膜炎 ) ,良性腹水 6例 (肝硬化 )。胸 /腹水离心后 ,细胞团用作端粒酶活性检测。如是血性胸 /腹水 ,加蒸馏水低渗破坏红细胞。检测端粒酶活性方法采用端粒重复序列扩增银染法 (PCR -TRAP) ,同时进行胸 /腹水脱落细胞学常规检查。结果 恶性胸 /腹水 40例中 35例端粒酶活性检测阳性 ,阳性率 87.5 % ;良性胸 /腹水 1 0例中仅 1例端粒酶检测阳性 ,阳性率 1 0 % ,恶性胸 /腹水端粒酶活性检测阳性率明显高于良性胸 /腹水 (P <0 .0 0 1 )。端粒酶活性诊断恶性胸 /腹水的敏感性为 87.5 %、特异性为 90 % ,高于细胞学检查阳性率 (55 % ) (P<0 .0 5)。恶性胸 /腹水端粒酶活性与肿瘤恶性程度有一定相关性 ,低分化肿瘤胸 /腹水端粒酶活性检测阳性率 96 .2 % ,而中高分化肿瘤71 .4% ,两者有显著差异 (P <0 .0 5)。结论 TRAP银染法可有效地检测到胸腹水脱落细胞的端粒酶活性。

Objective To evaluate the clinical significance of telomerase activity assay in differentiating whether the pleural fluid and ascite is malignant or not. Methods Fifty fresh specimens of pleural and peritoneal effusions were obtained from 50 patients with benign or malignant diseases, including 16 malignant pleural fluids (8 lung cancers, 5 breast cancers,3 lymphomas), 24 malignant ascites cases (3 primary hepatic carcinomas, 8 gastric carcinomas, 9 bowel cancer,4 lymphomas), 4 benign pleural fluids (tuberculous pleurisy cases), 6 benign ascites cases (liver cirrhosis cases). Fluid specimens were spun, a the residual cells were used for the telomerase activity assay. If the fluids were bloody, distilled water was added to the cell pellets to break the red blood cells. Telomerase activity was determined by telomeric repeat amplification protocol (TRAP). The results of cytology were collected in all the cases for comparison. Results Telomerase activity was detected in 35 (87.5%) of the 40malignant fluids, and 13 fluids with negative cytological results all presented positive results. Only 1 (10%) fluids were positive for telomerase activity in the 10 benign fluids. The positive rate of telomerase activity assay in malignant fluids was significantly higher than that in benign fluids (P<0.001), and also higher than cytological result (55%) (P<0.05).There was relation ship between the telomerase activity assay and the differentiation of primary tumour. Conclusion The TRAP assay can effiectively detect telomerase activity of the exfoliated cancer cells in the pleural fluid and ascite. The assay of telomerase activity for the diagnosis of malignant fluids is sensitive and specific and is useful for the differential diagnosis of benign and malignant fluids. There was relation ship between the telomerase activity assay and the differentiation of primary tumour. Telomerase could be a molecular marker for diagnosis of malignant pleural fluid and ascite,and it has potential as a diagnostic adjunct in cytopathology.