鳞状细胞癌抗原1 mRNA实时荧光定量RT-PCR检测方法建立与应用  

Establishment and application of the method to detect squamous cell carcinoma antigen 1 mRNA by real-time quantitative RT-PCR

作　　者：王晓楠[1,2,3,4] 程民 胡世莲[1,2,3,4] WANG Xiao-nan;CHENG Min;HU Shi-lian(Department of Geriatrics ,Anhui Provincial Hospital Affiliated to Anhui Medical University, Hefei 230001, P. R. China;Anhui Institute of Gerontology, He f ei 230001, P. R. China;Anhui Provincial Key Laboratory of Tumor Immunotherapy and Nutrition Therapy, Hefei 230001 ,P. R. China;Anhui Evidence-based Medicine Center, Hefei 230001 ,P. R. China)

机构地区：[1]安徽医科大学附属安徽省立医院老年医学科,安徽合肥230001 [2]安徽省老年医学研究所,安徽合肥230001 [3]肿瘤免疫与营养治疗安徽省重点实验室,安徽合肥230001 [4]安徽省循证医学中心,安徽合肥230001

出　　处：《中华肿瘤防治杂志》2018年第7期471-475,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：安徽省科技攻关项目(1501041142)

摘　　要：目的寻找特异性标志物,建立高效的检测方法对肺癌早期诊断尤为重要。本研究建立人鳞状细胞癌抗原1(squamous cell carcinoma antigen 1,SCCA 1)mRNA实时荧光定量RT-PCR检测方法,并评价其在肺鳞癌循环肿瘤细胞检测中的应用价值。方法收集安徽省立医院2014-02-01-10-31资料完整的肺鳞癌患者外周血标本28例(其中11例临床已确诊发生远处转移,17例临床未确诊转移),同时选取肺炎患者及正常人外周血标本各10例。运用荧光定量RT-PCR法检测上述样品中SCCA 1mRNA表达水平。采用回归分析方法制作定量标准曲线,阳性检出率比较采用多个独立样本的Kruskal-Wallis H检验。结果建立了外周血SCCA 1mRNA实时荧光定量RT-PCR检测方法,确定了引物、探针、标准品及对照品,检测方法的特异性强,灵敏度高。11例临床确诊已转移的肺鳞癌患者外周血SCCA 1mRNA均为阳性,17例临床未确诊转移的患者9例为阳性,8例为阴性,阳性率为71.4%,肺炎患者及正常人检测结果均为阴性,差异有统计学意义,χ2=36.456,P<0.05。结论建立的SCCA1mRNA实时荧光定量RT-PCR检测方法具有很高的临床应用价值,可用于检测肺鳞癌循环肿瘤细胞中SCCA1mRNA表达水平,可用于肺鳞癌患者的早期诊断。OBJECTIVE It＇s crucial to explore and establish method for specific markers of circulating tumor cell in lung cancer. The objective of this study was to investigate a quantitative method for detecting squamous cell carcinoma an- tigen 1 （SCCA 1） mRNA by real-time quantitative RT-PCR. The application of this method to detect the circulating tumor cells in the peripheral blood of lung squamous cell cancer patients will also be evaluated. METHODS Totally 28 cases lung cancer patients with peripheral blood samples,who were all in the Anhui Provincial Hospital between 2/1/2014 and 10/31/2014 and confirmed by pathological examination,were included in the study （11 cases were confirmed distal metas- tasis,17 cases not diagnosed metastasis）, 10 cases pneumonia patients, 10 healthy donors. RNA extraction and SCCA 1 mRNA were detected by the real-time quantitative RT-PCR in peripheral blood samples. Quantitative standard curve was calculated by regression analysis. The positive detection rate was performed by Kruskal-Wallis H test of multiple inde- pendent sample rates. RESULTS The method to detect the SCCA 1 mRNA using the real-time quantitative RT-PCR was successfully established by defining the primers, probes, standards and controls. The sensitivity and specificity of this method was high. All of the peripheral blood samples from 11 patients with metastatic lung cancer were positive for SCCA 1 mRNA,9 of 17 patients with non-metastatic lung cancer were positive for SCCA 1 mRNA in the peripheral blood samples,all of the blood samples from 10 pneumonia patients and 10 healthy donors were negative for SCCA 1 mRNA,which had statistically significant difference （χ^2=36. 456 ,P〈0. 05）. CONCLUSION The quantitative RT-PCR assay for SCCA 1 mRNA could be used for the detection of circulating tumor cells in the peripheral blood of lung cancer patients, which would be valuable for the diagnosis of lung squamous cell cancer.

关 键 词：肺鳞癌 循环肿瘤细胞 鳞状细胞癌抗原1 MRNA 逆转录聚合酶链反应 

分 类 号：R734.2[医药卫生—肿瘤]