中华医学遗传学杂志
中華醫學遺傳學雜誌
중화의학유전학잡지
CHINESE JOURNAL OF MEDICAL GENETICS
2009年
2期
151-155
,共5页
王玉川%徐晋珩%耿鑫%张维铭
王玉川%徐晉珩%耿鑫%張維銘
왕옥천%서진형%경흠%장유명
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
胃腫瘤%癌前病變%耑粒酶逆轉錄酶基因%選擇性剪接%聚閤酶鏈式反應
위종류%암전병변%단립매역전록매기인%선택성전접%취합매련식반응
gastric neoplasm%precancerous lesion%telomerase reverse transeriptase gene%alternative splicing%polymerase chain reaction
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 telomerasereverse 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),β位点保留的ASVs(α+β+γ+ASV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,B+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.
目的 檢測正常胃黏膜、胃癌前病變、胃癌組織中耑粒酶逆轉錄酶基因(human telomerasereverse transcriptase gene,hTERT)選擇性剪接變異體(alternative splicing variants gene,ASVs)的錶達,初步揭示胃癌多階段縯變過程中hTERT選擇性剪接模式的變化.方法 採用半巢式逆轉錄-PCR擴增8箇hTERT ASVs,瓊脂糖凝膠電泳檢測各ASVs的暘性率;應用SYBR Green實時定量逆轉錄-PCR法檢測胃癌和胃癌前病變組織中β+ASV的錶達水平.結果α+β+γ+ASV在正常胃黏膜中不錶達,在胃癌組織中暘性率比胃癌前病變高(P<0.05);β缺失型ASV在正常胃黏膜、胃癌前病變和胃癌組織中的暘性率分彆為72.2%、95.0%、100.0%(P>0.05),β位點保留的ASVs(α+β+γ+ASV缺失型ASV、γ缺失型ASV、αγ缺失型ASV)在正常胃黏膜、胃癌前病變、胃癌組織中的暘性率分彆為11.1%、40.0%、94.7%,3組間差異具有統計學意義(P<0.05).實時定量逆轉錄-PCR顯示.胃癌中β+ASV錶達水平比癌前病變高5.49倍.結論 胃癌多階段縯變過程中hTERT選擇性剪接模式不同,B+ASV在胃癌多階段縯變過程中錶達水平逐步升高,提示β+ASV的檢測可能為胃癌及胃癌前病變的診斷提供參攷依據.
목적 검측정상위점막、위암전병변、위암조직중단립매역전록매기인(human telomerasereverse transcriptase gene,hTERT)선택성전접변이체(alternative splicing variants gene,ASVs)적표체,초보게시위암다계단연변과정중hTERT선택성전접모식적변화.방법 채용반소식역전록-PCR확증8개hTERT ASVs,경지당응효전영검측각ASVs적양성솔;응용SYBR Green실시정량역전록-PCR법검측위암화위암전병변조직중β+ASV적표체수평.결과α+β+γ+ASV재정상위점막중불표체,재위암조직중양성솔비위암전병변고(P<0.05);β결실형ASV재정상위점막、위암전병변화위암조직중적양성솔분별위72.2%、95.0%、100.0%(P>0.05),β위점보류적ASVs(α+β+γ+ASV결실형ASV、γ결실형ASV、αγ결실형ASV)재정상위점막、위암전병변、위암조직중적양성솔분별위11.1%、40.0%、94.7%,3조간차이구유통계학의의(P<0.05).실시정량역전록-PCR현시.위암중β+ASV표체수평비암전병변고5.49배.결론 위암다계단연변과정중hTERT선택성전접모식불동,B+ASV재위암다계단연변과정중표체수평축보승고,제시β+ASV적검측가능위위암급위암전병변적진단제공삼고의거.
Objective To investigate the changes of the human telomerase reverse transcriptase gene (hTERT) aherative 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 mucosa, 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 mucosa (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 mueosa, 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.
