外周血中 Survivin 基因的检测及其临床意义

摘要

目的：建立一种特异性检测外周血中 Survivin 基因 mRNA 表达的逆转录双重荧光定量 PCR 方法。方法：设计特异性引物、荧光探针，检测健康人群和癌症患者治疗前后的 Survivin 基因表达量，并利用统计学方法对此基因的临床意义和治疗效果进行评价。结果：建立了 GADPH、Survivin 基因的标准曲线，并有良好的相关性，R2分别为0．9983和0．9960。管家基因 GADPH，在50例健康人群和47例癌症患者（治疗前后）体内表达量（分别为：6．45×105、6．57×105、6．74×105 copies／ml）无统计学差异。Survivin 基因，在健康人群中无表达（Ct 值≥28）；癌症患者（治疗前）检测到91．5％（43／47）表达，表达量为5．35×105 copies／ml；癌症患者（治疗后）检测到46．8％（22／47），表达量为3．18×105 copies／ml。结论：本研究建立了在外周血中检测 Sur-vivin 基因的双重荧光定量 PCR 方法，此方法在健康人群和癌症患者间、对癌症不同治疗方法的评估、对癌症治疗效果的评估等方面均有良好的应用。%Objective:To establish a specific detection of retroviral Survivin mRNA expression in peripheral blood of double FQ -PCR method.Methods:To design specific primers,fluorescent probes,Survivin gene expression before and after testing in healthy people and cancer patients was tested.Results:Established the standard curve of GADPH, Survivin gene,and there was a good correlation R2 were 0.998 3 and 0.996 0.The housekeeping gene GADPH,50 healthy people and 47 patients with cancer(after treatment)in vivo expression(respectively:6.45 ×105 ,6.57 × 105 ,6.74 ×105 copies/ml)was not statistically different.Survivin gene,was not expressed in healthy people(Ct val-ues≥28)in cancer patients(before treatment)detected in 9 1 .5 %(43 /47 )expression,the expression level of 5.35 ×105 copies/ml,cancer patients(after treatment)detected 46.8%(22 /47),the expression level of 3.18 × 105 copies/ml.Conclusion:This study was established to detect survivin dual fluorescence quantitative polymerase chain reaction(PCR)in peripheral blood,this method in healthy people and patients with cancer and evaluation of different methods for treating cancer,on the effectiveness of cancer treatment evaluation has a good application.