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目的:观察SMMC7721细胞在接受不同剂量60Coγ射线照射后线粒体DNA非编码区是否存在特定的断裂敏感点。方法:根据人线粒体DNA非编码区的基因序列设计相应的引物序列,再通过接头介导PCR(LMPCR)及基因扫描来检测其断裂程度和位点。结果:PCR产物电泳结果显示,接受不同剂量γ射线照射后,各剂量都有相同大小的片段。通过基因扫描的方法,发现不同照射剂量下的断裂位点位置是相同的。结果说明这些断裂敏感位点不是随机分布的。并且,位点的损伤频率也是随着剂量的增加而呈现递增趋势,随着剂量的增加还出现了新的断裂位点。结论:SMMC7721细胞线粒体DNA非编码区确实存在对γ射线敏感的位点。
OBJECTIVE: To observe whether SMMC7721 cells have specific cleavage-sensitive sites in the non-coding region of mitochondrial DNA after 60Coγ-ray irradiation at different doses. Methods: The corresponding primer sequences were designed according to the gene sequence of human non-coding region of mitochondrial DNA, and the degree of cleavage and site were detected by linker-mediated PCR (LMPCR) and gene scanning. Results: The electrophoresis results of PCR products showed that after receiving different doses of γ-ray irradiation, each dose had the same size of fragments. Through the method of gene scanning, it was found that the location of the cleavage sites was the same at different irradiation doses. The results show that these fracture-sensitive sites are not randomly distributed. Moreover, the site of injury frequency also showed an increasing trend with the dose increased, with the dose also increased the new fracture site. Conclusion: The mitochondrial DNA non-coding region of SMMC7721 cells do have some γ-ray sensitive sites.