不同分期胃癌患者外周血淋巴细胞Foxp3 mRNA表达水平的研究  

Differentially displayed Foxp3 mRNA in peripheral blood from patients with various stages of gastric cancer

作　　者：卢剑[1] 袁向亮[1] 沈立松[1] 

机构地区：[1]上海交通大学医学院附属新华医院检验科,上海200092

出　　处：《临床检验杂志》2007年第3期215-217,共3页Chinese Journal of Clinical Laboratory Science

摘　　要：目的采用SYBR GreenⅠ实时荧光定量PCR,研究不同分期胃癌患者外周血淋巴细胞Foxp3 mRNA的表达水平。方法普通RT-PCR结合胶回收制备Foxp3 mRNA-CDS标准品,利用标准品制备定量PCR标准曲线,对曲线进行灵敏度、精密度和特异性分析,用SYBR GreenⅠ实时荧光定量PCR检测40例胃癌和8例正常对照外周血Foxp3 mRNA。结果获得Foxp3全长CDS标准品,标准曲线范围(2.8×102)^(2.8×108)拷贝/μl,最低检测值280拷贝/μl,批内和批间变异系数(CV)<5%。Foxp3基因在胃癌患者外周血表达水平显著高于健康对照(P<0.01),表达水平与肿瘤分期显著相关(P<0.05)。结论Foxp3基因上调表达与胃癌的发生、发展和预后有关。Objective To discuss Foxp3 expression in peripheral blood from the patients with various stages of gastric cancer by SYBR Green Ⅰ real-time PCR assay. Methods Foxp3 standard was prepared by conventional RT-PCR and employed to generate quantitative PCR standard curve. The precision, sensitivity and specificity of the assay were measured. SYBR Green Ⅰ real-time PCR assay was adopted to detect and quantify Foxp3 expression in peripheral blood from 40 gastric cancer patients and 8 normal controls. Results The coding sequence （CDS） of full length Foxp3 was obtained and used as standards. The dynamic range of standard curve of real-time PCR assay was 10^2-10^8 copies. The mean coefficient of variation （CV） of inter- and intra-assay was less than 5%. The minimum detectable limit was 280 copies of Foxp3 mRNA. Foxp3 expression level in gastric cancer was significantly higher than that in normal control （P 〈 0.01 ）. In the patients with gastric cancer the expression level of Foxp3 significantly correlated to TNM staging. Conclusion The high expression of Foxp3 correlated to the development and prognosis of gastric cancer.

关 键 词：FOXP3 调节性T细胞 实时荧光定量PCR TNM分期 

分 类 号：R735.3[医药卫生—肿瘤]