人源Dickkopf-1原核表达产物在非小细胞肺癌患者血清中的应用研究

Study on application of human Dickkopf-1 prokaryotic expression products in serum of patients with non-small cell lung cancer

目的 探讨非小细胞肺癌患者血清中Dickkopf-1（DKK-1）自身抗体潜在的临床应用价值.方法 应用RT-PCR方法获得DKK-1 cDNA,构建原核表达载体pET30a（＋）/DKK-1,亲和层析方法纯化蛋白,SDS PAGE电泳及Western blot鉴定,应用间接ELISA方法对38名键康志愿者和97例非小细胞肺癌患者血清中 DKK-1自身抗体进行检测.结果 DKK-1自身抗体在非小细胞肺癌患者血清中的阳性率为34.0%,特异性为84.2%;DKK-1自身抗体与非小细胞肺癌患者有无远处转移存在相关性（P＝0.006）;DKK-1自身抗体和肿瘤标志物痛胚抗原联合检测的阳性率明显高于癌胚抗原与胃泌素释放肽前体、细胞角蛋白、神经元特异性烯醇化酶、鳞状细胞癌抗原进行联合检测的阳性率.结论 该研究成功构建原核表达载体pET30a（＋）/DKK 1.并建立检测DKK-1自身抗体的ELISA方法.DKK-1自身抗体可能成为非小细胞肺癌诊断、治疗中一种有用的肿瘤标志物.

Objective To investigate the potential clinical value of Dickkopf-1 （ DKK 1 ） autoantibody in serum of patients with non small cell lung cancer （NSCLC）. Methods DKK-1 cDNA was obtained by RT-PCR,prokaryotic expression vector pET30a （ ＋ ）/DKK-l was constructed,protein was purified by affinity chromalography. fractionated by sodium dodecyl sulfate polyacrylamide gel clectropheresis and appraised by Western blot. DKK-1 autoantibody in scrum of 38 healthy controls and 97 patients with NSCLC was detected by indirect enzyme linked immunosorbent assay. Results The positive rate of DKK-1 autoantibody in serum of patients with NSCLO was 34.0%. and the specificity was 84. 2%. DKK-1 autoantibody was correlated with distant metastasis of tumor in patients with NSCLC （ P = 0.006）. The positive rate of carcinoembryonic antigen combined with DKK-1 autoantibody was significantly higher than that with other tumor markers including pro-gastrin releasing peptide,cytokeratin,neuron-specific enolase.and squamous cell carcinoma antigen. Conclusions In our research,the prokaryotic expression vector pET30a （ ＋ ）/DKK 1 is successfully constructed,and indirect enzyme linked immunosorbent assay is applied to detect DKK-1 autoantibody in serum of patients with NSCLC. DKK 1 autoantibody may become a useful tumor marker in the diagnosis and treatment of NSCLC.