非小细胞肺癌患者血清中SP70的检测及其临床意义

A preliminary study on serum protein SP70 as a novel biomarker for the detection of non-small cell lung cancer

目的评价血清中单抗NJ001相关靶抗原SP70能否成为检测非小细胞肺癌（NSCLC）的新型血清肿瘤标志物。方法以SPC—A1为抗原免疫新西兰兔，制备多抗。用多抗包被微孔板，NJ001为一抗，酶标记的羊抗鼠抗体为标记抗体，建立双抗体夹心ELISA法，并优化实验条件；采用该法对2007年10月至2011年6月从南京医科大学第一附属医院收集的经病理确诊的175例肺癌（包括80例肺腺癌、70例肺鳞癌和25例小细胞肺癌）、25例肺良性疾病患者和300名健康体检者血清中SP70进行检测；同时与电化学发光法检测癌胚抗原（CEA）、神经元特异性烯醇化酶（NSE）和角质蛋白19片段（CYFRA21—1）进行比较；各组问阳性率比较采用，检验或Fisher确切概率法。结果肺腺癌、肺鳞癌、小细胞肺癌和肺良性疾病组的阳性率分别为68．8％、51．4％、16．0％和12．0％，均明显高于健康体检组（7．3％）；NSCLC组（肺腺癌和肺鳞癌）阳性率高于小细胞肺癌组（60．7％vs16．0％，x2=17．23，P〈0．05）和肺良性疾病组（60．7％vs12．0％，r=20．41，P〈0．05）；68例有分期资料的NSCLC患者中，早期组（x2=30）的阳性率达76．7％。另外，ELISA法对NSCLC患者SP70总检出率高于CEA、NSE和CYFRA21—1（32．7％、18．0％和37．3％），差异有统计学意义（60．7％vs32．7％，χ^2=23．63，P〈0．05；60．7％vs18．0％,χ^2=57．22，P〈0．05；60．7％vs37．3％，χ^2=16．34，P〈0．05）。结论NSCLC患者血清中SP70阳性检出率较高，SP70可能是潜在的用于诊断NSCLC的血清肿瘤标志物。

Objective To evaluate whether the protein SP70 could be used as a serum biomarker for the diagnosis of non-small cell lung cancer （NSCLC）. Methods Polyclonal antibody was prepared by immunizing New Zealand rabbit with SPC-A1 cells. Sandwich ELISA was carried out by using newly-prepared polyclonal antibody（ PcMb ）coating assay plates, monoclonal antibody （McAb）NJ001 and HRP goat anti- mouse antibody as primary antibody and labeling antibody respectively. After optimizing the experiment conditions, serum from 175 lung cancer patients [ 80 NSCLC adenocarcinoma, 70 NSCLC squamous carcinoma and 25 small cell lung cancer（SCLC） ] ,25 benign lung disease（BLD） patients and 300 healthy controls （HC） were examined. CEA, NSE, CYFRA21-1 were measured by ECLIA for comparison. Results Positive rates of NSCLC adenocareinoma, NSCLC squamous carcinoma, SCLC and BLD were 68.8% , 51.4%, 16.0% and 12.0% respectively, obviously higher than that of HC （7.3%）. NSCLC （adenocarcinoma, Squamous carcinoma） had significantly higher positive rate than SCLC （60. 7% vs 16.0% ,X2 = 17. 23, P 〈 0. 05 ） and BLD （60. 7% vs 12. 0% ,X2 = 20. 41, P 〈 0. 05 ）. Among 68 NSCLC patients who had definite staging, positive rates at early stage（ 1/11, n=30） reached up to 76.7%. Meanwhile,positive rates of CEA, NSE and CYFRA21-1 （ 32. 7% , 18.0% and 37.3% ） were significantly lower than the targeting antigen to McAb NJ001 in NSCLC（60. 7% vs 32. 7% ,X2 =23.63 ,P 〈0.05 ;60. 7% vs 18.0%,X2 =57.22,P〈0.05;60.7% vs 37.3%,X2 = 16.34,P〈0.05）. Conclusions It showed high positive rates of SP70 in the serum of NSCLC patients, which suggested that SP70 might be a potential valuable biomarker for the diagnosis of NSCLC.