X射线辐照引起的线粒体D310片段突变的快速检测

张昕; 周鑫; 张红; #

doi:10.11804/NuclPhysRev.28.01.118

X射线辐照引起的线粒体D310片段突变的快速检测

A Novel Approach for Rapid Detection of Xray Irradiation Induced mtDNA D310 Mutation

摘要

摘要: 采用4 Gy X射线辐照人乳腺癌细胞MCF7，分别在照射后0， 2， 4， 8， 16， 24， 48， 72和144 h时间点收集细胞，提取DNA并以聚合酶链式反应进行扩增，再以BsaXI限制性内切酶对扩增产物进行消化，消化产物以琼脂糖凝胶电泳进行分离。结果表明， 4 Gy X射线辐照可引起D310片段突变，且该突变在经辐照之后144 h检测水平明显超过野生型。 Human breast cancer cell line MCF7 was irradiated with 4 Gy Xray， collected at 0， 2， 4， 8， 16， 24， 48， 72, 144 h after irradiation， respectively. Whole genome DNA including mtDNA were extracted at each time point， and amplified by polymerase chain reaction(PCR). Then the PCR product was subjected to BSAXI digestion， all of digestion product then underwent a brief electrophoresis. Results showed D310 mutation can be induced by 4 Gy Xray irradiation and D310 mutation can overwhelm the normal phenotype 144 h after irradiation.

Abstract: Human breast cancer cell line MCF7 was irradiated with 4 Gy Xray， collected at 0， 2， 4， 8， 16， 24， 48， 72, 144 h after irradiation， respectively. Whole genome DNA including mtDNA were extracted at each time point， and amplified by polymerase chain reaction(PCR). Then the PCR product was subjected to BSAXI digestion， all of digestion product then underwent a brief electrophoresis. Results showed D310 mutation can be induced by 4 Gy Xray irradiation and D310 mutation can overwhelm the normal phenotype 144 h after irradiation.

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