外周血叶酸受体阳性循环肿瘤细胞检测在非小细胞肺癌筛查中的应用价值

Peripheral blood folate receptor-positive circulating tumor cells detection in the screening of non-small cell lung cancer

目的观察非小细胞肺癌患者外周血叶酸受体阳性循环肿瘤细胞(circulating tumor cells,CTCs)水平变化,探讨外周血叶酸受体阳性CTCs在非小细胞肺癌筛查中的应用价值。方法非小细胞肺癌患者136例为肺癌组,肺部良性病变患者10例为良性病变组,健康志愿者54例为对照组。3组均采用以叶酸受体为靶点的免疫磁珠阴性富集+实时荧光定量PCR法检测外周血叶酸受体阳性CTCs水平;采用化学发光免疫分析法检测血清癌胚抗原(carcino-embryonic antigen,CEA)、糖链抗原(carbohydrate antigen,CA)125、CA724、细胞角蛋白19片段(cytokeratin 19fragment,CYFRA21-1)、神经元特异性烯醇化酶(neuron-specific enolase,NSE)水平。比较3组CTCs水平;比较不同临床特征非小细胞肺癌患者CTCs水平;绘制ROC曲线,评价CTCs及血清CEA、CA125、CA724、CYFRA21-1、NSE 5项指标联合诊断非小细胞肺癌的价值。结果肺癌组CTCs水平[11.21(8.58,15.30)FU/3mL]高于良性病变组[7.55(5.23,10.25)FU/3mL]和对照组[4.95(3.55,7.62)FU/3mL](P<0.05),良性病变组高于对照组(P<0.05)。TNM分期Ⅰ、Ⅱ、Ⅲ、Ⅳ期非小细胞肺癌患者CTCs水平[11.00(8.58,13.30)、13.25(10.48,16.88)、14.77(11.47,16.55)、17.89(17.07,19.22)FU/3mL]两两比较差异均有统计学意义(H=16.443,P<0.05);不同年龄、性别、肿瘤最大径、T分期、分化等级、病理类型非小细胞肺癌患者CTCs水平比较差异均无统计学意义(P>0.05)。ROC曲线分析结果显示,CTCs以8.70FU/3mL为最佳截断值,诊断非小细胞肺癌的AUC为0.953(95%CI:0.926~0.979,P<0.05),灵敏度为79.40%,特异度为98.10%,诊断效能优于CEA、CA125、CA724、CYFRA21-1、NSE联合检测。结论非小细胞肺癌患者外周血叶酸受体阳性CTCs水平升高,且增高程度与TNM分期有关;外周血叶酸受体阳性CTCs可用于非小细胞肺癌的早期筛查。

Objective To investigate the application value of peripheral blood folate receptor-positive circulating tumor cells(CTCs)detection to the screening of non-small cell lung cancer(NSCLC)by observing the changes of CTCs in NSCLC patients.Methods The CTCs levels were detected by tumor specific folate ligand oligonucleotide conjugates and real-time fluorescence quantitative PCR in 136patients with NSCLC(NSCLC group),10patients with benign lung diseases(benign group)and 54healthy volunteers(control group).The serum carcino-embryonic antigen(CEA),carbohydrate antigen(CA)125,CA724,cytokeratin 19fragments(CYFRA21-1)and neuron-specific enolase(NSE)were detected by chemiluminescence immunoassay.The levels of CTCs were compared among groups,and among patients with different clinical features.ROC was drawn to assess the values of CTCs combined with serum CEA,CA125,CA724,CYFRA21-1and NSE to the diagnosis of NSCLC.Results The level of CTCs was higher in NSCLC group(11.21(8.58,15.30)FU/3 mL)than that in benign group(7.55(5.23,10.25)FU/3 mL)and control group(4.95(3.55,7.62)FU/3mL)(P<0.05),and higher in benign group than that in control group(P<0.05).Significant statistic difference was found in multiple comparison in the level of CTCs among TNM stageⅠ,Ⅱ,ⅢandⅣ(11.0(8.58,13.30),13.25(10.48,16.88),14.77(11.47,16.55),17.89(17.07,19.22)FU/3mL)(H=16.443,P<0.05).There were no significant differences in CTCs level among different ages,genders,maximum diameters of tumors,T stages,differentiated degrees and pathological types(P>0.05).When the optimal cut-off value of CTCs was 8.70 FU/3mL,the AUC was 0.953(95%CI:0.926-0.979,P<0.05),the sensitivity was 79.40%,and the specificity was 98.10%.The diagnostic efficacy of CTCs was superior to the joint detection of CEA,CA125,CA724,CYFRA21-1and NSE.Conclusion The level of folate receptor-positive CTCs in peripheral blood increases in NSCLC patients,and is correlated with TNM stage.The detection of peripheral blood folate receptor-positive CTCs can be used in the early screening of NSCLC.