肿瘤坏死因子相关凋亡诱导配体对肿瘤细胞PRDXs表达影响的观察

Effect of tumor necrosis factor-related apoptosis-inducing ligand on PRDXs expression

目的:探讨肿瘤坏死因子相关凋亡诱导配体(TRAIL)对肿瘤细胞中过氧化氧化还原蛋白(PRDXs)表达的影响及其机制。方法:选取肿瘤细胞,设立对照组和TRAIL处理组;用实时定量PCR法和蛋白质印迹法检测PRDXs在各种肿瘤细胞中的表达;用PCR法克隆PRDX4启动子,用萤光素酶含量测定其含量。结果:与对照组相比,TRAIL处理组中PRDX1、PRDX2、PRDX3、PRDX5和PRDX6mRNA及蛋白表达差异无统计学意义,P>0.05;PRDX4mRNA和蛋白的表达显著降低,P<0.01;放线菌素D处理8h时,TRAIL处理组与对照组中PRDX4mRNA降解近50%,差异无统计学意义,P>0.05;TRAIL显著降低pPRDX4/-979的活性呈剂量依赖方式。结论:TRAIL在转录水平上下调肿瘤细胞中PRDX4基因的表达,对PRDX4mRNA的稳定性没有影响。

OBJECTIVE： To investigate the mechanism of Peroxiredoxins （PRDXs） expression in the panel of cancer cells affected by tumor necrosis factor-related apoptosis-inducing ligand（TRAIL）. METHODS： A panel of cancer cells were selected and set up control group and TRAIL treatment groups for investigation; the expression of PRDXs mRNA and protein in the panel of cancer ceils was confirmed by real-time RT-PCR and Western blot, respectively; Generation of PRDX4 promoter subcloned by PCR. The luciferase activity was determined using the dual-luciferase reporter assay sys- tem. RESULTS.. TRAIL significantly reduced PRDX4 mRNA and protein levels （P〈 0. 01）, while PRDX1, PRDX2, PRDX3,PRDX5 and PRDX6 had no obvious effects （P〉0.05） when compared with control group; PRDX4 mRNA in HeLa treated with control or TRAIL degraded approximately by 50% after 8 h treatment with actinomycin D,there was no significant difference between two groups（P〉0.05）. TRAIL significantly decreased pPRDX4/_979 activity in a dose- dependent manner. CONCLUSION： TRAIL suppressed PRDX4 expression at the transcriptional level, have no effect on PRDX4 mRNA stability.