定量RT-PCR测定食管鳞癌组织中人叉头样转录因子3基因的表达

Measurement of the Forkhead Box P3 Gene Expression Levels in Esophageal Squamous Cell Carcinoma by Real-time Quantitative Reverse Transcription-polymerase Chain Reaction

目的测定食管鳞状细胞癌组织中人叉头样转录因子3(forkhead box P3,FOXP3)的基因表达水平,为食管癌的免疫治疗提供依据。方法基于TaqMan荧光探针技术,构建克隆载体pMD 18-T-FOXP3,作为定量模板,建立检测FOXP3信使核糖核酸(mRNA)含量的实时荧光定量逆转录-聚合酶链反应(RT-PCR)方法,在GeneAmp7500型检测仪上测定42例食管鳞癌组织和30例正常对照组织中FOXP3 mRNA的含量。结果食管鳞癌组织FOXP3 mRNA表达高于正常对照组织[(72.20±23.10)×104拷贝/μg RNA vs.(0.68±0.34)×104拷贝/μg RNA;P<0.05]。结论成功建立了人FOXP3基因mRNA表达含量的荧光定量检测方法,FOXP3在食管鳞癌组织中的表达较正常对照组织增高。

Objective To detect the expression of forkhead box P3 （FOXP3）gene in esophageal squamous cell carcinoma（ESCC） and provide a new basis for immunotherapy of esophageal cancer. Methods Based on fluorescent TaqMan methodology, a real-time quantitative reverse transcription-polymerase chain reaction （RT-PCR） for detecting the expression of FOXP3 was set up. In this method, a cloning vector DMD 18-T-FOXP3 was constructed as a standard plasmid. The specific expression of FOXP3 in 42 patients with ESCC and 30 healthy controls were measured by using GeneAmp 7500 Sequence Detection Systems. Results FOXP3 mRNA copy number in ESCC was significantly higher than that in healthy control tissue [（72.20±23.10） × 10^4copy/μg RNA vs. （0.68±0.34） × 10^4copy/μg RNA;P〈0.05]. Conclusion A real-time quantitative RT-PCR method for detecting the expression of FOXP3 gene in ESCC has been successfully established. The expression level of FOXP3 is increased in ESCC compare with healthy controls.