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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

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【作者】 张海燕孟欣都镇先邓娓娓王华芹

【Author】 ZHANG Hai-yan1,MENG Xin2,DU Zhen-xian1,DENG Wei-wei1,WANG Hua-qin2 1.First Affiliated Hospital,China Medical University,Shenyang 110001,P.R.China 2.Department of Biochemistry and Molecular Biology,China Medical University,Shenyang 110001,P.R.China

【机构】 中国医科大学附属第一医院老年病干诊科中国医科大学基础医学院生化与分子生物教研室中国医科大学附属第一医院内分泌科

【摘要】 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。

【Abstract】 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 cells 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 system.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.

【基金】 辽宁省科技攻关项目(2010225032);辽宁省教育厅资助项目(2009A765)
  • 【文献出处】 中华肿瘤防治杂志 ,Chinese Journal of Cancer Prevention and Treatment , 编辑部邮箱 ,2013年06期
  • 【分类号】R730.2
  • 【被引频次】2
  • 【下载频次】153
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