不同分期胃癌患者外周血淋巴细胞Foxp3 mRNA表达水平的研究

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（2.8×10^2）～（2.8×10^8）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.

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

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 102-108 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.