^(60)Coγ射线对人线粒体COXI和ATPase6表达水平的影响

Effects of ^(60)Co Gamma Rays on Mitochondrial COXI and ATPase6 Expression in Human Peripheral Blood

目的观察60Coγ射线对人外周血线粒体COXI和ATPase6基因和蛋白质水平表达的影响,探讨线粒体编码基因用于辐射生物剂量估算的可行性。方法采用60Coγ射线照射正常人离体外周血,照射剂量率为1.013 Gy/min,剂量分别为0、0.2、0.4、0.6、0.8、1.0、2.0、3.0、4.0、5.0 Gy,。照后6小时,用RT-PCR技术检测基因的表达水平,用流式细胞仪检测蛋白质的表达水平,分别拟合最佳回归方程,建立剂量-效应曲线。结果(1)0~3.0 Gy,COXI基因表达水平的最佳回归方程为Y=0.629+0.256D(r2=0.920,p<0.01),ATPase6基因表达水平的最佳回归方程为Y=0.221+0.805D(r2=0.912,p<0.01)。(2)0~5.0 Gy,COXI蛋白质表达水平的最佳回归方程为Y=1.054+0.595D(r2=0.919,p<0.01);ATPase6蛋白质表达水平与照射剂量间的关系不明显,不能拟合出最佳回归方程。结论60Coγ射线照射人离体外周血,随着照射剂量的增加,线粒体COXI的基因与蛋白质表达水平一致,呈增加趋势,且有良好的线性关系,有望用于辐射生物剂量的估算。

This paper is to explore the effect of 60 Co gamma rays on mitochondrial gene and protein expression in human Peripheral blood .Human peripheral blood samples were exposed to 60 Co gamma rays at different dos-es ：0 .0 ,0 .2 ,0 .4 ,0 .6 ,0 .8 ,1 .0 ,2 .0 ,3 .0 ,4 .0 ,5 .0 Gy ,after 6 hours irradiation in human peripheral blood ,Changes of mRNA level of COXI and ATPase 6 genes were detected by RT-PCR and Real-Time PCR . And COXI and ATPase6 protein expression change was observed by using flow cytometer .The relationship be-tween mitochondrial gene and protein expression with dose was analyzed by origin 7 .5 software .The expression of two genes at mRNA level was up-regulated ,A good dose-effect relationship was showed at dose range of 0-3 Gy for COXI （Y=0 .629＋0 .256 D ,r 2=0 .9169 ,p〈0 .01） .At the protein level ,A good dose-effect rela-tionship was showed for COXI （Y=1 .054＋0 .595 D , r2 =0 .919 ,p〈0 .01 ） at dose range of 0-5 Gy .The expression of COXI at mRNA and protein level were not noticeable ,dose-effect relationship between protein expression of ATPase6 with dose was not significan .The expression of COXI at mRNA and protein level were up-regulated and accorded ,COXI could be promising radiation biological dosimeter .