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狄春红, 谭晓华, 仙玲玲, 顾少华, 杨军, 周婷, 周迪, 杨磊. 硫氧还蛋白还原酶2基因抗砷作用[J]. 中国公共卫生, 2009, 25(1): 56-58. DOI: 10.11847/zgggws2009-25-01-26
引用本文: 狄春红, 谭晓华, 仙玲玲, 顾少华, 杨军, 周婷, 周迪, 杨磊. 硫氧还蛋白还原酶2基因抗砷作用[J]. 中国公共卫生, 2009, 25(1): 56-58. DOI: 10.11847/zgggws2009-25-01-26
DI Chun-hong, TAN Xiaa-hua, XIAN Ling-ling, . Study on arsenic tolerance of thioredoxin reductase 2 gene in arsenic resistance ECV 304 cells[J]. Chinese Journal of Public Health, 2009, 25(1): 56-58. DOI: 10.11847/zgggws2009-25-01-26
Citation: DI Chun-hong, TAN Xiaa-hua, XIAN Ling-ling, . Study on arsenic tolerance of thioredoxin reductase 2 gene in arsenic resistance ECV 304 cells[J]. Chinese Journal of Public Health, 2009, 25(1): 56-58. DOI: 10.11847/zgggws2009-25-01-26

硫氧还蛋白还原酶2基因抗砷作用

Study on arsenic tolerance of thioredoxin reductase 2 gene in arsenic resistance ECV 304 cells

  • 摘要: 目的 研究硫氧还蛋白还原酶2(thioredoxin reductase,TrxR2)基因在抗砷细胞(As-ECV304)中的抗砷作用。方法 蛋白印迹法(Western Blot)检测As-ECV304和对照组细胞TrxR2蛋白水平,流式细胞仪检测细胞周期分布,化学合成TrxR2基因小干扰RNA(small interfering RNA,siRNA)转染As-ECV304细胞,Western Blot验证干扰效率,细胞急性毒性试验检测细胞抗砷性的改变。结果 As-ECV304细胞中TrxR2蛋白水平较对照组细胞高,As-ECV304细胞G0/G1期细胞所占比例高于对照细胞。3个siRNA中有一个能特异性抑制TrxR2基因表达,干扰组细胞的生存率低于阴性对照组和未干扰组,3组细胞的半数抑制率IC50分别为9.13,15.88和18.52μmol/L。结论 siRNA干扰TrxR2基因表达后能明显降低抗砷细胞的抗砷性,提示该基因在细胞抗砷机制中发挥了重要作用。

     

    Abstract: Objective To study the function of thioredoxin reductase 2(TrxR2)gene in a rsenic-resistance(As- ECV 304)cells.Methods The prote in level of TrxR2 in As-ECV 304 and control ECV 304 cells was detected by Wes tern Blot and the cell cycles were detected by flowcy tometry(FCM).Three short in terfering RNA(siRNA)of TrxR2 gene and negative controls iRNA were made from chemical synthesis.As-ECV 304 cells transfected with 3 TrxR2 siRNA and control siRNA were exposed to NaAsO2 for 72 hours and the effect of RNA in terference was studied by Western Blot.As-ECV 304 Cells transfected secondly with the TrxR2 siRNA,control siRNA and un-transfected were cultured for 24 hours and then exposed to NaAsO2 for 24 hours.Cell viability and a rsenic to lerance were analyzed by MTT assay.Results Western Blot showed that the TrxR2 prote in level was highe rin As-ECV 304 cells than in control ECV 304 Cells.Cell cycle of As- ECV 304 cell and control cell showed different percent in phase and the percent of G0/G1 phase was higher in As-ECV 304 cells.One of three TrxR2 siRNA can specially in terfered target gene expression.MTT assay showed that the cell viability was obviously decreased in cells transfected with TrxR2 siRNA than control siRNA.The 50%inh ib iting concentration (IC50)of As-ECV 304 cells transfected with TrxR2 siRNA,control siRNA and without transfection exposed to NaAsO2 was 9.13μmol/L,15.8μmol/L and 18.52μmol/L,respectively.Conclusion The arsenic to lerance of As-ECV 304 cells transfected with TrxR2 siRNA significantly decreased,the TrxR2 gene may play an importan trole in arsenic to lerance of As-ECV 304 cells.

     

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