CAJ | 학술논문

目的：研究肠三叶因子(intestinal trefoil factor，ITF)对自身启动子活性的影响及Janus激酶/信号转导和转录激活因子(Janus kinase/signal transducers and activators of transcription，JAK/STAT)信号通路对其的调控作用。方法：以人全血基因组DNA为模板，PCR获取ITF基因5'侧翼序列，插入pGL3-Basic载体，构建ITF启动子重组载体；以不同浓度的ITF刺激，检测双荧光素酶的活性；运用JAK-STAT3通路特异性抑制剂AG490阻断JAK-STAT3信号通路，观察AG490对ITF启动子活性的影响。结果：酶切和直接测序法证实包含ITF启动子的重组质粒构建成功；瞬时转染实验显示，ITF能显著提高ITF启动子的活性(P<0.05)；加入AG490后ITF启动子活性显著下降(P<0.05)。结论：ITF通过JAK-STAT3途径上调自身启动子的活性。
목적：연구장삼협인자(intestinal trefoil factor，ITF)대자신계동자활성적영향급Janus격매/신호전도화전록격활인자(Janus kinase/signal transducers and activators of transcription，JAK/STAT)신호통로대기적조공작용。방법：이인전혈기인조DNA위모판，PCR획취ITF기인5'측익서렬，삽입pGL3-Basic재체，구건ITF계동자중조재체；이불동농도적ITF자격，검측쌍형광소매적활성；운용JAK-STAT3통로특이성억제제AG490조단JAK-STAT3신호통로，관찰AG490대ITF계동자활성적영향。결과：매절화직접측서법증실포함ITF계동자적중조질립구건성공；순시전염실험현시，ITF능현저제고ITF계동자적활성(P<0.05)；가입AG490후ITF계동자활성현저하강(P<0.05)。결론：ITF통과JAK-STAT3도경상조자신계동자적활성。
Objective: To investigate the effect ofintestinal trefoil factor (ITF) on the transcriptional activity of ITF promoter and to explore the regulatory mechanismofJanus kinase/signal transducersand activators of transcription(JAK/STAT) on ITF promoter. Methods: The 5' flanking sequence of the ITF gene was cloned from human whole blood genomic DNA by PCR. ITF promoter fragment was cloned and inserted into the pGL3-Basic vector to construct recombinant vector. ITF promoter vector was stimulated with ITF at various concentrations and the luciferase activity was measured. The JAK-STAT3 signal transductionpathway was then blocked by a speciifc inhibitor AG490 to determine the signal pathway involved in ITF promoter activity. Results: Restriction endonuclease analysis and DNA sequencing confirmed that the recombinant plasmid, containing ITF promoter, was constructed successfully. After transient transfection, the activity of ITF promoter was increased signiifcantly in the presence of ITF (P<0.05). Blockage of the JAK-STAT3 signal transduction pathway with AG490 signiifcantly reduced the ITF promoter activity (P<0.05). Conclusion: ITF increases the transcriptional activity of ITF promoter via the JAK-STAT3 signal transduction pathway.