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目的 探讨FasL对CD2 8基因启动子活性的影响,为研究凋亡信号在免疫衰老中的作用提供资料。方法 采用MTT法和annexinⅤ- FITC染色测定细胞凋亡,Northernblot检测CD2 8mRNA的表达水平,构建CD2 8基因上游非编码区序列的报告基因载体,并瞬时转染JurkatE6 - 1细胞后,用Luciferase检测报告基因启动子的活性。结果 FasL在JurkatE6 -1细胞介导的凋亡可使CD2 8mRNA水平降低。CD2 8启动子的有效序列在上游- 4 0 0bp的范围内,并且FasL介导的凋亡信号可明显降低CD2 8启动子活性。结论 FasL介导的凋亡信号是CD2 8基因启动子活性降低的重要原因。
Objective To investigate the effect of FasL on the promoter activity of CD28 gene in order to provide information for studying the role of apoptosis signaling in immune senescence. Methods Apoptosis was determined by MTT assay and Annexin V-FITC staining. The expression of CD28 mRNA was detected by Northern blot. The reporter gene vector was constructed and sequenced by Luciferase assay after transient transfection of JurkatE6 - 1 cells. Gene promoter activity. Results FasL mediated apoptosis in JurkatE6-1 cells decreased the level of CD28 mRNA. The effective sequence of the CD28 promoter is in the upstream-400 bp range, and the FasL-mediated apoptotic signal significantly reduces the CD28 promoter activity. Conclusion FasL-mediated apoptosis signal is an important reason for the decrease of CD2 8 gene promoter activity.