| 小鼠肿瘤线粒体DNA突变研究 |
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| 作者姓名： | Dai JG Min JX Zhang GQ Wei H Xiao YB |
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| 作者单位： | 1. 400037,重庆,第三军医大学新桥医院胸外科
2. 第三军医大学实验动物中心 |
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| 基金项目： | 国家自然科学基金资助项目 ( 3 990 0 173 ),第三军医大学校科研基金资助 |
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| 摘 要： | 目的探讨线粒体DNA(mtDNA)突变与肿瘤发生发展的关系。方法采用聚合酶链反应(PCR)技术结合限制性片段长度多态性分析(PCR—RFLP)和单链构象多态性分析(PCR—SSCP)技术了解Lewis肺癌、LA795、Hca—F、Hca—P、CT26、SCC891共6个小鼠肿瘤细胞系mtDNA的基因变异。结果经PCR—RFLP分析发现,这些肿瘤细胞系mtDNA编码区的tRNA^lle Glu Met及ND1基因核酸序列,在27个限制性内切酶酶切位点上均无差异。而在非编码区(D-loop),与对照小鼠相比,Hca—F出现了Hinf Ⅰ的新酶切位点;用PCR—SSCP分析方法对这些肿瘤细胞系mtDNA的D—loop的5’及3’端作进一步分析,在6个肿瘤细胞系中,有4个在mtDNA非编码区上存在着突变。结论D—loop是肿瘤细胞mtDNA突变的高发区,mtDNA突变在肿瘤的发生发展中可能起作用。
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| 关 键 词： | 肿瘤细胞系 小鼠肿瘤 线粒体DNA突变 非编码区 Lewis肺癌 mtDNA突变 ND1 酶切位点 核酸序列 PCR—SSCP分析 |
mtDNA mutations in mouse tumors |
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| Dai JG,Min JX,Zhang GQ,Wei H,Xiao YB.mtDNA mutations in mouse tumors[J].Chinese Journal of Pathology,2004,33(5):458-461. |
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| Authors: | Dai Ji-gang Min Jia-xin Zhang Guo-qiang Wei Hong Xiao Ying-bin |
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| Institution: | Department of Thoracic Surgery, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China. |
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| Abstract: | Objective To investigate variations of mtDNA in mouse tumors and to explore the relationship between mtDNA mutations and murine carcinogenesis. Methods Variations of D-loop, ND3 and tRNA~(Ile Glu Met) gene fragments of mtDNA from six mouse tumor cell lines were analyzed by PCR-RFLP and PCR-SSCP techniques. Results ND3 and tRNA~(Ile Glu Met) gene fragments of mtDNA from the tumors showed no variations at 27 endonuclease sites. The D-loop of mtDNA from Hca-F demonstrated an additional endonuclease site of Hinf I in contrast to the inbred mouse. Upon PCR-SSCP analysis, the D-loop of mtDNA was found to possess mutations in 4 of 6 tumors. Conclusion D-loop appears to be the hot spot for tumor mtDNA mutations, which may contribute to the carcinogenesis of murine tumors. |
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| Keywords: | Tumor cells cultured DNA mitochondrial Mutations |
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