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目的研究人热休克因子1(HSF1)对Fas配体(FasL)启动子转录活性的影响。方法用在线启动子分析软件分析FasL启动子区转录因子结合位点;凝胶阻滞实验(EMSA)研究内源性HSF1与FasL启动子区的热休克元件(HSE)结合能力;将组成型活化的HSF1突变体与FasL启动子表达载体FasL-luc共同转染HeLa细胞,采用双荧光素酶报告基因检测系统,检测荧光索酶活性。结果在FasL启动子区发现HSE核心序列(nGAAnnTTCn),HSF1可以与该HSE体外结合;荧光素酶活性测定发现HSF1明显上调FasL-luc转录活性,突变该HSE,则转录激活作用消失。结论 HSF1对FasL具有转录调控作用。
Objective To investigate the effect of human heat shock factor 1 (HSF1) on the transcriptional activity of Fas ligand (FasL) promoter. Methods The transcription factor binding sites of FasL promoter region were analyzed by using online promoter analysis software; the heat shock component (HSE) binding ability of endogenous HSF1 and FasL promoter region was studied by EMSA; constitutive activation HSF1 mutant was co-transfected with FasL promoter expression vector FasL-luc into HeLa cells. Fluorescein was detected by dual luciferase reporter assay. Results The HSE core sequence (nGAAnnTTCn) was found in FasL promoter region. HSF1 could be bound to this HSE in vitro. Luciferase activity assay showed that HSF1 up-regulated the FasL-luc transcription activity. Mutation of this HSE resulted in the disappearance of transcriptional activation. Conclusion HSF1 has a transcriptional regulatory effect on FasL.