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1、血清hTERTmRNA实时荧光PCR定量方法的建立及初步应用刘阳[通信作者]刘阳，Email:liuyang081@gmail.com，电话：（0731）88639075，胡梅2，吴娅娜3喻红波1（410006长沙，1武警湖南总队医院检验科2湖南中医药大学生物化学教研室3湖南中医药大学图书馆）[摘要]目的建立血清人端粒酶逆转录酶（humantelomerasereversetranscriptase,hTERT）mRNA实时荧光定量PCR检测方法，并初步应用于胃癌检测。方法以hTERT基因为靶基因设计
2、引物，构建携带hTERTcDNA的质粒pMD-hTERT，建立血清hTERTmRNA实时荧光PCR定量方法，检测48例胃癌患者，58例健康人血清端粒酶逆转录mRNA表达水平并分析其差异。结果建立血清hTERTmRNA表达的实时荧光定量PCR方法，胃癌患者hTERTmRNA相对表达水平显著高于健康人（p<0.01），与肿瘤大小和分期、分化等级明显相关（p<0.01），与年龄、性别、以及肿瘤数量无明显关联（p＞0.05）。结论本研究成功建立了血清hTERTmRNA表达的实时荧光定量PCR方法，实验表明检测
3、血清hTERTmRNA有助于胃癌诊断和研究。[关键词]人端粒酶逆转录酶，实时荧光定量PCR，胃癌[中图法分类号][文献标识码]Real-timefluorescencequantitativePCRassayforhumanTelomerasereversetranscriptasemRNAinserumandpreliminaryapplicationinGastriccancerLiuYang1,HuMei,WuYana,YuHongbo(1，4DepartmentofClinicalLabora
4、tory,HunanProvincialCorpsHospital,ChinesePeople’sArmedPoliceForces,2,3HunanUniversityofTraditionalChineseMedicine,Changsha,410006,China)[Abstract]ObjectiveToestablishreal-timefluorescencequantitativepolymerasechainreactionmethodfordetectinghumantelomera
5、sereversetranscriptasemRNAinserum,andstudieditsclinicalsignificanceingastriccancerpatients.MethodsDesignedhTERTprimersandconstructedplasmidpMD-hTERTcarryinghTERTcDNA,andestablishedaquantitativePCRassaytodetecthTERTmRNAexpressionlevelsinserumof48gastricc
6、ancerpatientsand58healthyindividuals.ResultsAreal-timefluorescencequantitativePCRmethodfordetectinghTERTmRNAlevelsinserumwasestablished,TheexpressionlevelofhTERTinserumofgastriccancergroupwassignificantlyhigherthanhealthygroup.Therelativeexpressionlevel
7、ofhTERTmRNAwasdemonstratedtobeindependentlycorrelatedwithtumorsize,gradeandTNMstage,whilenotcorrelatedwithage,genderandnumberoftumors.ConclusionThereal-timequantitativePCRfordetectingserumhTERTmRNAwasestablishedsuccessfullywiththecharacterofhighspecific
8、ity,sensitivityandrepeatability,whichmightprovideanoveldiagnosisandresearchtoolforgastriccancer.[KeyWords]humantelomerasereversetranscriptase，real-timefluorescencequantitativepolymerasechainreaction，Gastriccancer端粒酶在hTERT的作用下延长随细
