摘要
目的探讨过氧化氢对PUMA启动子活性的调控作用。方法采用基因克隆技术构建荧光素酶报告基因表达质粒,利用荧光素酶报告基因检测技术检测不同处理因素下启动子的活性。结果在肠癌LoVo细胞中,过氧化氢可以诱导PUMA启动子的活性,去除Sp1结合位点后,过氧化氢处理后PUMA启动子的活性下降,但与转染PUMA全启动子组的活性并没有显著性差异(P=0.132);而在去除了p53和Sp1转录结合位点后,启动子活性较去除了Sp1结合位点后明显下降,差别有显著性意义(P=0.000)。结论在肠癌LoVo细胞中,过氧化氢可以刺激PUMA启动子的活性,Sp1转录因子在这一过程中起到一定的辅助作用,Sp1通过与p53协同效应而发挥作用。
Objective To investigate the mechanism of H2O2-induced PUMA promoter activity in colorectal cancer cells.Methods Promoter reporter gene plasmid was constructed and transfected into colorectal cancer cells and luciferase assay was used to detect the activity of PUMA promoter after treatment of H2O2 or inhibitors.Results H2O2 stimulated the activity of a 493 PUMA promoter.Deletion of the Sp1-binding sites also decreased the transactivation of PUMA promoter by H2O2,but there was no significant difference.Furthermore,induction of PUMA promoter activity by H2O2 was abrogated by PFT-α(a p53 inhibitor)and Mithramycin A(a Sp1 inhibitor)as compared with PFT-α alone.Conclusion Our findings suggest that Sp1 works together with p53 in the regulation of H2O2-induced PUMA promoter activity.
出处
《现代消化及介入诊疗》
2010年第2期74-77,共4页
Modern Interventional Diagnosis and Treatment in Gastroenterology