热休克转录因子1对高迁移率族蛋白B1的转录调控作用及机制

The Transcription Regulation and Molecular Mechanism of HSF1 on HMGB1

目的:探讨烧伤血清诱导下热休克转录因子1(HSF1)对高迁移率族蛋白B1(HMGB1)的转录调控作用及其机制。方法:构建热休克转录因子1真核表达载体pcDNA3.1-HSF1,HMGB1野生型启动子荧光素酶报告基因pGL3-HMGB1-Y,HMGB1突变型启动子荧光素酶报告基因pGL3-HMGB1-T。pcDNA3.1-HSF1转染巨噬细胞RAW264.7,烧伤血清诱导后,半定量RT-PCR检测HMGB1 mRNA的表达。pcDNA3.1-HSF1,HMGB1启动子荧光素酶报告基因共转染RAW264.7,烧伤血清诱导后,检测并比较pGL3-HMGB1-Y和pGL3-HMGB1-T的相对萤光素酶活性。结果:烧伤血清诱导下,HSF1可以下调RAW264.7 HMGB1mRNA的表达。pGL3-HMGB1-T的相对荧光素酶值较pGL3-HMGB1-Y明显下降,P<0.01,差异有显著统计学意义。结论:烧伤血清诱导下,HSF1可能通过与HMGB1启动子区HSE的结合下调小鼠巨噬细胞RAW264.7 HMGB1 mRNA的表达。

Objective：To explore the transcription regulation and molecular mechanism of Heat Shock Transcription Factor 1 （ HSF1 ） on HMGB1 in RAW264.7 induced by burn serum. Methods： Constructing eukaryotic expression vector containing mouse HSF1 pcDNA3. 1-HSF1 , luciferase reporter gene containing mouse HMGB1 wild type promoter pGL3-HMGB1-Y, and luciferase reporter gene containing mouse HMGB1 mutant type promoter pGL3-HMGB1-T. The mRNA expressions of HMGB1 was determined with Sq RT-PCR after pcDNA3.1-HSF1 transfected RAW264.7 induced by burn serum, peDNA3. 1-HSF1 and the luciferase reporter gene containing mouse HMGB1 promoter transfected RAW264.7 together, then the relative lueiferase activity of pGL3-HMGB1-Y and pGL3-HMGB1-T were determined and compared. Results：HSF1 can downregulate the expressions of HMGB1 mRNA in RAW264.7 induced by burn serum.The relative luciferase activity of pGL3-HMGB1-T is lower significantly than that of pGL3-HMGB1 -Y, P 〈 0.01, there is statistical significance. Conclusion： HSF1 can downregulate the expression of HMGB1 mRNA in RAW264.7 induced hy burn serum, which might be affected by the binding of HSFI to the HSE in the promoter of HMGBI.