| 陈俊生,赵麟,李静,等.基于荧光共振能量转移技术的STAT3二聚化抑制剂筛选模型的建立[J].中国海洋药物,2018,37(3):9-17. |
| 基于荧光共振能量转移技术的STAT3二聚化抑制剂筛选模型的建立 |
| Establishment%20of%20the%20screening%20model%20for%20STAT3%20dimerization%20inhibitor%20based%20on%20fluorescence%20resonance%20energy%20transfer%20technology |
| 投稿时间：2017-12-19 修订日期：2018-03-05 |
| DOI： |
| 中文关键词: 信号传导与转录激活因子3(STAT3) 二聚化 荧光共振能量转移(FRET) Stattic |
| English Keywords:Signal%20transducers%20and%20activators%20of%20transcription%203(STAT3) %20dimerization %20fluorescence%20resonance%20energy%20transfer%20(FRET) %20Stattic. |
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| 中文摘要: |
| 目的%20构建STAT3二聚化抑制剂筛选模型,为STAT3抑制剂筛选提供实验方法。方法%20分别构建pECFP-N1-STAT3和pEYFP-N1-STAT3的荧光报告载体,利用脂质体转染技术将二者共转入HEK-293T细胞,利用ECFP和EYFP两种荧光蛋白之间能量共振转移,检测磷酸化STAT3分子的二聚化水平,并检测Stattic对二聚化的影响。结果%20成功构建了pECFP-N1-STAT3和pEYFP-N1-STAT3的荧光报告载体。将两种荧光报告载体共转染HEK-293T细胞后,Western%20Blot%20检测结果显示STAT3以及p-STAT3的表达水平明显增加。以458nm波长激发ECFP,其发射波长可激发EYFP。加入STAT3二聚化抑制剂Stattic后,荧光强度降低,且呈现一定的剂量依赖性。结论%20基于荧光共振能量转移技术的STAT3二聚化抑制剂筛选模型构建成功。 |
| English Summary: |
| Abstract:%20Objective to construct%20a%20screening%20model%20of%20STAT3%20dimerization%20inhibitor%20and to provide%20a%20novel%20experimental%20method%20for%20STAT3%20inhibitor%20screening.%20Methods%20Two%20fluorescence%20reporter%20vectors,%20pECFP-N1-STAT3%20and%20pEYFP-N1-STAT3,%20were%20constructed%20by%20molecular%20biological%20technique;%20Liposome-mediated%20transfection%20technique%20was%20used to transfer%20gene%20into%20HEK-293T%20cells;%20The%20mutual%20function%20between%20ECFP%20and%20EYFP%20was%20confirmed%20using%20the%20energy%20resonance%20transfer%20technology%20by%20which%20the%20dimerization%20level%20of%20STAT3%20proteins%20was%20measured;The%20effect%20of%20Stattic%20on%20dimerization%20of%20STAT3%20proteins%20was%20examined%20in%20this%20model.%20Results%20The%20fluorescent%20reporter%20vectors%20containing%20the%20STAT3%20gene%20were%20successfully%20constructed.%20Western%20Blot%20showed%20that%20the%20expression%20levels%20of%20STAT3%20and%20phosphorylated%20STAT3%20proteins%20were%20significantly%20increased%20after%20transfection.%20The%20ECFP%20was%20excited%20at%20a%20wavelength%20of%20458%20nm%20and%20its%20emission%20wavelength%20could%20excite%20EYFP.%20The%20addition%20of%20STAT3%20dimerization%20inhibitor%20Stattic%20induced%20the%20reduction%20of%20fluorescence%20intensity%20in%20dose-dependent%20manner,%20suggesting%20that%20Stattic%20inhibited%20STAT3%20dimerization%20in%20this%20model.%20Conclusion%20The%20screening%20model%20of%20STAT3%20dimerization%20inhibitor%20is%20established%20successfully%20based%20on%20FRET%20technology. |
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