OLC1蛋白在非小细胞肺癌患者外周血血浆中的水平及其临床意义

摘要

Objective The aim of this study was to detect the plasma concentration of OLC 1 ( overexpressed in lung cancer 1 ) protein as a potential cancer biomarker , and evaluating its clinical application value in the diagnosis of non-small cell lung cancer (NSCLC).Methods We prepared OLC1 antibody with OLC1 full length protein, in 5-6-week old Bal B/c mice.Each mouse was immunized four times at a dose of 15-30μg antigen protein , and the interval between two consecutive immunizations was two weeks.Antibody screening was made by ELISA and Western blot , and a double antibody sandwich ELISA kit was developed .We used this established ELISA kit to detect the plasma concentration of OLC 1 protein in 281 NSCLC patients and 92 gender-and age-matched healthy controls .Area under the receiver operating characteristic curve (AUC) was used to evaluate the detection efficacy of OLC 1.Results We obtained 11 OLC1 monoclonal antibodies and successfully established the ELISA kit to detect the plasma concentration of OLC1 with a detection range from 1.95 ng/ml to 62.50 ng/ml.OLC1 concentration in the case group (124.69 ng/ml) was significantly higher than that in the control group (67.07 ng/ml, P<0.001).In the scenario of distinguishing NSCLC from control group , AUC result was 0.69.When the cut-off was set at 67.72 ng/ml, the sensitivity and specificity was 84.4%and 51.1%, respectively.In term of distinguishing early lung cancer (IA) from normal controls, the AUC, sensitivity and specificity were 0.68, 77.8% and 54.4%, respectively .Conclusion The plasma concentration of OLC 1 protein is significantly elevated in NSCLC patients.OLC1 may be as a potential cancer biomarker applied in clinical diagnosis .%目的：探讨肺癌相关蛋白OLC1在非小细胞肺癌（NSCLC）患者外周血血浆中的水平及其临床意义。方法取5～6周龄Bal B/c小鼠，OLC1全长蛋白作为抗原免疫制备OLC1抗体。每只小鼠免疫4次，每间隔2周免疫1次，每次15～30μg抗原蛋白。通过Western blot方法筛选抗体，制备检测血浆中OLC1蛋白的双抗体夹心法酶联免疫吸附试验（ ELISA）试剂盒，并检测281例NSCLC患者和92例健康对照者（性别年龄匹配）血浆中OLC1蛋白的水平。采用受试者工作特征曲线（ ROC曲线）下面积评估OLC1对NSCLC的检测效能。结果共制备了11株特异性好的OLC1单克隆抗体，并成功建立了检测血浆OLC1的ELISA试剂盒，检测范围为1．95～62．50 ng/ml。 NSCLC患者外周血血浆中位OLC1蛋白水平（124．69 ng/ml）显著高于健康对照者（67．07 ng/ml，P＜0．001）。 OLC1区分NSCLC和健康对照者的ROC曲线下面积为0．69（95％CI为0．62～0．76），当OLC1的cut off值为67．72 ng/ml时，其灵敏度为84．4％，特异度为51．1％。 OLC1区分早期（ⅠA期） NSCLC和健康对照者的ROC曲线下面积为0．68（95％CI为0．6～0．77），其灵敏度为77．8％，特异度为54．4％。结论NSCLC患者血浆中OLC1蛋白水平显著升高，可成为辅助临床诊断的候选标志物。