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目的检测和分析转移特性不同的两个小鼠肝癌细胞亚系线粒体DNA(mtDNA)的遗传变异,探讨线粒体DNA遗传改变与肿瘤发生发展的关系。方法PCR-RFLP和序列测定技术。结果对mtDNA的tRNAIle+GlN+Met基因和ND3基因以及D-loop片段进行的扩增和限制性片段长度多态性分析结果显示,无扩增片段长度呈多态性,且这两个肝癌细胞系mtDNA的所有限制性片段方式和大小完全一致。序列测定发现,这两个肝癌细胞系在线粒体DNA的D-loop区存在序列差异。结论mtDNA非编码区内的遗传改变,反映了肿瘤发生发展过程中环境和遗传因素的影响,有可能与肿瘤细胞的恶性表型有关。
Objective To detect and analyze the genetic variation of mitochondrial DNA (mtDNA) in two murine hepatocellular carcinoma cell lines with different metastatic potential and to explore the relationship between the genetic alteration of mitochondrial DNA and tumorigenesis. Methods PCR-RFLP and sequence determination techniques. Results The analysis of mtDNA tRNAIle + GlN + Met gene, ND3 gene and D-loop fragment and restriction fragment length polymorphism showed that there was no polymorphism in the length of amplified fragments, and the two hepatocellular carcinoma cells All restriction fragments of mtDNA are exactly the same size and size. Sequencing revealed that there were sequence differences in the D-loop region of mitochondrial DNA between the two hepatoma cell lines. Conclusion The genetic changes in the non-coding region of mtDNA reflect the environmental and genetic factors in the process of tumorigenesis, which may be related to the malignant phenotype of tumor cells.