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[目的]探讨人survivin核心启动子的位置及在A549细胞中的转录激活功能。[方法]在生物信息学分析基础上,构建了4个分别由survivin 5′上游-1436~+64、-612~+64、-414~+64和-203~+64片段驱动的报告载体,分别瞬时转染A549及HK-2细胞,观察其在两种细胞的表达活性并比较在A549细胞中表达的活性差异。[结果]生物信息学提示survivin TSS位于ATG上游-64bp处,启动子近端富含若干潜在Sp1、p53和HIF-1α等结合位点。4种报告载体在A549细胞均有活性,且不同长度的启动子活性存在统计学差异。而在HK-2细胞中,4种载体未见显著性差异。[结论]人survivin启动子的活性具有肿瘤特异性,核心启动子可能位于TSS上游-203bp至+64bp处。suvivin核心启动子内可能存在2处HIF-1α的结合位点。
[Objective] To investigate the location of the human survivin core promoter and its transcriptional activation in A549 cells. [Method] On the basis of bioinformatics analysis, four reporter vectors were constructed, which were driven by survivin upstream of -1436 ~ +64, -612 ~ +64, -414 ~ +64 and -203 ~ +64 respectively. The A549 and HK-2 cells were transiently transfected respectively, and their activity in both cell lines was observed. The differences in the activity of A549 cells were also observed. [Results] Bioinformatics suggested that survivin TSS was located at -64bp upstream of ATG, and the proximal promoter was rich in some potential sites of Sp1, p53 and HIF-1α. The four kinds of reporter vectors were all active in A549 cells, and the promoter activity of different length had statistical difference. In HK-2 cells, there was no significant difference among the four vectors. [Conclusion] The activity of human survivin promoter is tumor-specific, and the core promoter may be located at -203 bp to +64 bp upstream of TSS. There may be two HIF-1α binding sites in suvivin core promoter.