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电离辐射对肺腺癌细胞A549线粒体的影响 被引量:1

Effects of ionizing radiation on the mitochondrion in adenocarcinoma A549 cells
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摘要 目的 研究电离辐射对肺腺癌细胞A549线粒体的影响.方法 0、0.5、3和8 Gy60Co γ射线照射A549细胞后,利用JC-1探针检测照射后肺腺癌细胞A549线粒体膜电位的变化;使用ATP试剂盒检测辐射对A549细胞ATP生成的影响;利用实时定量PCR方法检测线粒体拷贝数.结果 在照射后24 h,各剂量组A549细胞膜电位出现明显改变(F=243.44,P<0.05),与0 Gy照射组相比,0.5和3 Gy照射的A549细胞线粒体膜电位显著升高(t=-10.12、-5.59,P<0.05),而8 Gy照射的线粒体膜电位显著降低(t=15.22,P<0.05),照后48 h各照射组线粒体膜电位基本恢复正常(F=10.36,P<0.05);照射后24 h ATP水平与膜电位变化趋势一致(F=97.08,P<0.05),其中0.5和3 Gy A549细胞中ATP水平升高(t=1.66、7.27,P <0.05),8 Gy照射组降低(t=-8.24,P<0.05);照射后48 h,ATP水平也发生变化(F =39.49,P<0.05),0.5和3 Gy照射后A549细胞中ATP水平显著高于0 Gy组(=4.60、8.53,P<0.05).照射后24 h线粒体拷贝数显著增加,其中0.5 Gy mtDNA拷贝数增加至对照组的12倍(t=0.02,P<0.05),3和8 Gy组mtDNA拷贝数分别增加至对照组的7和10倍(=9.68、15.10,P<0.05).结论 大剂量电离辐射会导致A549细胞线粒体膜电位降低,从而影响ATP的生成,中等剂量以下的照射会导致A549细胞线粒体膜电位代偿性增高,从而使ATP的生成增多,8 Gy以内的电离辐射可使mtDNA拷贝数代偿性升高. Objective To investigate the effect of radiation on the mitochondrion in adenocarcinoma A549 cells.Methods After A549 cells were irradiated with 0,0.5,3 or 8 Gy of 60Co γrays,mitochondrion membrane potential of A549 cells was detected by JC-1 probe,and ATP activity was measured by ATP kit in a chemiluminescence apparatus.The mitochondria DNA copy numbers was detected by real-time PCR assay.Results At 24 h after radiation,the mitochondrion membrane potential of A549 cells in all the irradiated groups changed significantly (F =243.44,P < 0.05),among which 0.5 or 3 Gy of radiation resulted in a significant increase of mitochondrion membrane potential of A549 cells (t =-10.12,-5.59,P < 0.05).However,the mitochondrion membrane potential of A549 cells exposed to 8 Gy of radiation decreased significantly 24 h after radiation (t =15.22,P < 0.05).The mitochondrion membrane potential of A549 cells in all radiation groups returned to the normal level 48 h after radiation (F =10.36,P < 0.05).24 h after radiation,the level of ATP in A549 cells significantly changed respectively(F =97.08,P < 0.05),similar to the mitochondrion membrane potential.The ATP level in 0.5 and 3 Gy groups increased significantly (t =1.66,7.27,P < 0.05),and the level of ATP in 8 Gy group decreased significantly (t =-8.24,P < 0.05).Furthermore,48 h after both 0.5 and 3 Gy of radiation,the ATP content in A549 cells was still higher than that in untreated A549 cells (t =4.60,8.53,P <0.05).The mitochondria DNA copy numbers in A549 cells increased significantly in all the radiation groups (F =118.00,P < 0.05).Compared with untreated A549 cells the mitochondria DNA copy numbers in A549 cells increased at 0.5 Gy by 12 times(t =0.02,P <0.05),and increased at 3 and 8 Gy by 7 and 10 times,respectively (t =9.68,15.10,P < 0.05).Conclusions High dose of radiation resulted in the decrease of mitochondrion membrane potential of A549 cells,which subsequently affected the production of ATP.However,radiation with moderate and lower dose could lead to the compensatory increase of mitochondrion membrane potential of A549 cells,which promoted the production of ATP.The mitochondria DNA copy numbers compensatory would increase after A549 cells were exposed to radiation within 8 Gy.
出处 《中华放射医学与防护杂志》 CAS CSCD 北大核心 2014年第2期88-90,94,共4页 Chinese Journal of Radiological Medicine and Protection
基金 国家自然科学基金(30870749,81001216,31340051,81172593)
关键词 辐射 线粒体膜电位 线粒体拷贝数 三磷酸腺苷 Radiation Mitochondrion membrane potential Mitochondrion copy ATP
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