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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 establish a real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) method to detect the expression of hTERT mRNA in gastric cancer and to explore the relationship between the expression of hTERT and gastric cancer and its value in the diagnosis of gastric cancer. Methods Real-time quantitative detection of hTERTmRNA expression in 35 cases of gastric cancer and its corresponding marginal tissues was performed by Taqman technique and LightCycler fluorescence quantitative PCR instrument. The 10-fold of the hTERT to GAPDH copy number ratio was used as normalized hTERT (NhTERT). Results (1) The NhTERT in gastric cancer and their corresponding marginal tissues were 6.27 ± 0.89 and 0.93 ± 0.18 respectively, with significant difference between the two groups (t = 12.76, P <0. 0 1). (2) The expression level of hTERT mRNA in gastric cancer tissues was closely related to the degree of differentiation (P <0.01), but not with age, sex, tumor size, location and pTNM stage. Conclusion Real-time fluorescence quantitative RT-PCR method can accurately and efficiently quantify hTERT mRNA. Real-time quantitative detection of hTERTmR NA may contribute to the early diagnosis of gastric cancer.