摘要
目的 检测正常胃黏膜、胃癌前病变、胃癌组织中端粒酶逆转录酶基因（human telomerase reverse transcriptase gene，hTERT）选择性剪接变异体（alternative splicing variants gene，ASVs）的表达，初步揭示胃癌多阶段演变过程中hTERT选择性剪接模式的变化。方法 采用半巢式逆转录-PCR扩增8个hTERT ASVs，琼脂糖凝胶电泳检测各ASVs的阳性率；应用SYBR Green实时定量逆转录-PCR法检测胃癌和胃癌前病变组织中β^＋ ASV的表达水平。结果 α^＋β^＋γ^＋ASV在正常胃黏膜中不表达，在胃癌组织中阳性率比胃癌前病变高（P％0．05）；β缺失型ASV在正常胃黏膜、胃癌前病变和胃癌组织中的阳性率分别为72．2％、95．0％、100．0％（P〉0．05）；8位点保留的ASVs（α^＋β^＋γ^＋ASV、α缺失型ASV、γ缺失型ASV、αγ缺失型ASV）在正常胃黏膜、胃癌前病变、胃癌组织中的阳性率分别为11．1％、40．0％、94．7％，3组间差异具有统计学意义（P〈0．05）。实时定量逆转录-PCR显示，胃癌中β^＋ ASV表达水平比癌前病变高5．49倍。结论 胃癌多阶段演变过程中hTERT选择性剪接模式不同，β^＋ ASV在胃癌多阶段演变过程中表达水平逐步升高，提示β^＋ASV的检测可能为胃癌及胃癌前病变的诊断提供参考依据。
Objective To investigate the changes of the human telomerase reverse transcriptase gene （hTERT） alterative splicing pattern in gastric carcinogenesis. Methods Three alternative splicing sites （α, β,γ） were selected to design primers. The expression of eight hTERT alternative splicing variants （ASVs） in normal gastric mueosa, precancerous lesions and gastric cancer was detected by semi-nested reverse transcription-polymerase chain reaction （RT-PCR）. The expression of β site-remaining ASV （β^＋ hTERT mRNA） in precancerous lesions and gastric cancer tissues was detected by SYBR green real-time RT-PCR. Results The positive rate of α^＋β^＋γ^＋ hTERT mRNA was significantly higher in gastric cancer than in precancerous lesions and normal mueosa （94.7% vs. 40.0% and 0, P〈0.05）. The positive rates of other ASVs were not different among the three groups. The positive rates of β-deletion ASV were 72. 2% in normal mucosa, 95. 0% in precancerous lesions and 100. 0% in gastric cancer. The mRNA level of β^＋ hTERT was 5. 49 folds higher in gastric cancer than in precancerous lesions. Conclusion The hTERT alternative splicing pattern changes during gastric carcinogenesis. The β^＋ hTERT mRNA is expressed increasingly during gastric carcinogenesis and may provide useful information for diagnosis of gastric cancer or precancerous lesions.
出处
《中华医学遗传学杂志》
CAS
CSCD
北大核心
2009年第2期151-155,共5页
Chinese Journal of Medical Genetics
基金
国家自然科学基金（39770298）
关键词
胃肿瘤
癌前病变
端粒酶逆转录酶基因
选择性剪接
聚合酶链式反应
gastric neoplasm
precancerous lesion
telomerase reverse transcriptase gene
alternative splicing
polymerase chain reaction
