摘要
目的 建立一种实时荧光逆转录多聚酶链反应 (RT PCR) ,检测喉鳞状细胞癌组织人端粒酶逆转录酶 (hTERT)mRNA表达水平 ,并与传统的RT PCR进行比较。方法 采用TriZOL提取总RNA并将mRNA逆转录成cDNA ,然后利用TaqMan技术定量检测喉鳞状细胞癌组织hTERTmRNA。结果 实时荧光RT PCR检测癌组织的阳性率为 97.0 6 % ,癌旁组织的阳性率为 38.2 4 % ,差异显著 (χ2 =2 4 .2 5 ,P <0 .0 0 5 ) ,与癌旁组织比较 ,喉鳞状细胞癌组织hTERTmRNA表达水平平均上升 17.88倍 ;传统的RT PCR检测喉鳞状细胞癌组织和癌旁组织的阳性率分别为 73.5 3%和 11 77% ,均显著低于实时荧光RT PCR的阳性率 (P <0 .0 1)。结论 实时荧光RT PCR是检测喉癌组织hTERTmRNA水平的一种快速有效、灵敏度和特异性更高的方法 ,hTERTmRNA表达的上调可能是导致喉鳞状细胞癌发生的重要因素。
Objective To establish a real time fluorescent RT-PCR for quantitative determination of human telomerase reverse transcriptase mRNA in laryngeal squamous cell carcinoma and to compare the results to those of conventional RT-PCR.Methods Total RNA was extracted with TriZOL and mRNA was transcribed reversely into cDNA,then Taqman technique was used to quantitate hTERT mRNA in laryngeal squamous cell carcinoma tissue.Results 1.By determination with real time fluorescent RT-PCR,the positive rate of hTERT mRNA was 38.23% in adjacent normal tissue and 97.05% in laryngeal squamous cell carcinoma tissue respectively and there was a significant difference between carcinoma and adjacent normal tissue(χ2=24.25,P<0.005).Comparing to adjacent normal tissue,the expression levels of hTERT mRNA on the average was 17.88-fold increase in laryngeal squamous cell carcinoma tissue.2.By determination with conventional RT-PCR,the positive rate was 42.65% in adjacent normal tissue and laryngeal squamous cell carcinoma tissue and there was a significant difference between the positive rate detected with real time fluorescent RT-PCR and conventional RT-PCR(χ2=7.61,0.01>P>0.005).Conclusion The real time fluorescent RT-PCR is a rapid,sensitive and very specific method for quantitating human telomerase.The upregulation of hTERT mRNA may play an important role during the occurrence of laryngeal squamous cell carcinoma.
出处
《临床检验杂志》
CAS
CSCD
北大核心
2004年第5期335-337,共3页
Chinese Journal of Clinical Laboratory Science
基金
湖北省科技攻关项目 ( 2 0 0 2AA3 0 4B10 )
