胃癌中人端粒酶逆转录酶mRNA的实时定量检测  

Quantification of human telomerase reverse transcriptase mRNA in gastric cancer with real-time fluorescent quantitative RT-PCR

作　　者：胡丽华[1] 陈凤花[1] 李一荣[1] 王琳[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院,武汉430022

出　　处：《中华实验外科杂志》2004年第12期1482-1484,共3页Chinese Journal of Experimental Surgery

基　　金：湖北省科技攻关项目 (2 0 0 2AA30 4B1 0 )

摘　　要：目的　建立一种实时荧光定量逆转录 聚合酶链反应 (RT PCR)方法 ,检测胃癌组织中hTERTmRNA的表达 ,探讨hTERT的表达水平与胃癌的关系及其在胃癌诊断中的价值。方法采用Taqman技术与LightCycler荧光定量PCR仪对 3 5例胃癌及其相应切缘组织中hTERTmR NA的表达进行实时定量检测。将hTERT与GAPDH拷贝数之比的 10 0倍作为标准化hTERT(NhTERT)。结果　 (1)胃癌及其相应切缘组织中NhTERT分别为 6.2 7± 0 .89、0 .93± 0 .18,两者之间差异有统计学意义 (t =12 .76,P <0 .0 1)。(2 )胃癌组织中hTERTmRNA的表达水平与组织的分化程度密切相关 (P <0 .0 1) ,而与患者的年龄、性别、肿瘤的大小、定位以及 pTNM分期无相关性。结论　实时荧光定量RT PCR方法能对hTERTmRNA进行准确、高效的定量。hTERTmR NA的实时定量检测可能有助于胃癌的早期诊断。Objective To set up a real time fluorescent quantitative RT PCR assay,to detect human telomerase reverse transcriptase (hTERT) mRNA in gastric carcinomas,and to analyze the correlation between the expression level of hTERT mRNA and gastric carcinomas,and to evaluate the value of quantitative determination of hTERT mRNA in the diagnosis of gastric carcinomas.Method A real time quantitative RT PCR (RQ RT PCR) assay based on TaqMan fluorescence methodology and the LightCycler system was used to quantify the full range of hTERT mRNA copy numbers in 35 samples of gastric carcinomas and corresponding adjacent non cancerous tissues.The normalized hTERT (NhTERT) was standardized by quantifying the number of GAPDH transcripts as internal control and expressed as 100×(hTERT/GAPDH) ratio.Results (1) NhTERT from gastric carcinomas and corresponding adjacent non cancerous tissues was 6.27± 0.89 and 0.93± 0.18, respectively ( t = 12.76, P < 0.001). (2) There was no significant association between gastric cancer hTERT mRNA expression level and patient's age,gender,tumor size,location and tumor stage (PTNM),but a significant correlation between hTERT mRNA expression level in gastric carcinomas and the degree of differentiation was found ( P = 0.007). Conclusion Quantitative determination of hTERT mRNA by RQ RT PCR is a rapid and sensitive method.hTERT may have potential as a biomarker for the early detection of gastric cancer.

关 键 词：胃癌 人端粒酶逆转录酶 MRNA 实时检测 定量检测 荧光定量逆转录-聚合酶链反应 

分 类 号：R735.2[医药卫生—肿瘤]