胸腔积液中白细胞对端粒酶活性影响的分析

Research about the influence of white blood cells on telomerase activity in pleural effusion

目的通过免疫磁珠技术去除胸腔积液中的白细胞(WBC)后检测其中肿瘤细胞的端粒酶活性,探讨白细胞对胸腔积液肿瘤细胞端粒酶活性的影响。方法 收集15例胸腔积液,其中良性胸腔积液5例,恶性胸腔积液10例,应用TRAP-PCR-ELISA法检测分别经免疫磁珠阴性法及密度梯度离心法富集后胸腔积液细胞端粒酶活性。结果 利用吸光度差值(ΔA)定性分析显示去除白细胞前后良恶性胸腔积液端粒酶活性无明显差异(P>0.05);利用患者胸腔积液肿瘤细胞相对端粒酶活性值(RTA)定量分析,10例恶性胸水经两种方法富集后的RTA值比较有差异(P<0.05),免疫磁珠阴性法富集后恶性胸水肿瘤细胞RTA明显高于密度梯度离心法富集后的恶性肿瘤细胞;5例良性胸水经两种方法富集后的RTA值之间比较无显著性差异(P>0.05)。结论 定量分析显示利用免疫磁珠阴性法富集胸腔积液肿瘤细胞后进行端粒酶活性检测去除了白细胞对肿瘤细胞端粒酶活性的影响,提高了检测的敏感性和特异性,可用于良恶性胸腔积液的鉴别。

[ Abstract] Objective To clarify the influence of white blood cells （WBC） on telomerase activity in pleural effusions, using immunomagnetic negative enrichment method to remove WBC and enrich tumor cells of pleural effusions. Methods Experiments specimens were collected from patients with benign or malignant plenraJ effusions, including 5 benign samples and 10 malignant samples. Tumor cells were enriched from pleural effusion samples by means of the immunomagnetic beads negative enrichment method and density gradient centrifugation method, and then TRAP- PCR-ELISA method was used to examine telomerase activity of tumor cells enriched by the two methods. Results Using delta absorbance qualitative analysis, there was no significant difference in benign and malignant pleural effusions before and after removing WBC （ P 〉 0.05 ）. However, using relative telomerase activities （RTA） quantitative analysis, the telomerase activity of malignant pleural effusions after negative method enrichment was higher than they enriched by gradient centrifugation（P 〈0.05）. In 5 benign pleural effusion samples, there was no significant difference between the two methods （ P 〉 O. 05 ）. Conclusions The detection of tumor ceils telomerase activity after negative method enrichment depleted the influence of WBC to tumor cells＇ telomerase activity, and improves the detection sensitivity and specificity. It is beneficial to distinguish malignant pleural effusions from benign ones.