天津医药

• 细胞与分子生物学 • 上一篇    下一篇

线粒体DNA D-loop基因多态性与肾透明细胞癌关系的研究

张胜雷,徐金升,白亚玲,张俊霞,崔立文,张慧然   

  1. 河北医科大学第四医院
  • 收稿日期:2013-07-30 修回日期:2013-11-14 出版日期:2014-03-15 发布日期:2014-03-15
  • 通讯作者: 张胜雷

Relationship between single nucleotide polymorphisms in Mitochondrial Displacement-loop and the risk of renal cell carcinoma

  • Received:2013-07-30 Revised:2013-11-14 Published:2014-03-15 Online:2014-03-15

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摘要： 目的 探讨线粒体DNA D-loop（mtDNA D-loop）基因多态性与肾透明细胞癌发病风险的关系。方法 应用病例对照研究，采用聚合酶链反应（PCR）对59例肾透明细胞癌患者和68例健康对照的mtDNA D-loop区进行扩增并测序。将测序结果与线粒体文库中的Revised Cambridge Reference Sequence（rCRS)比对分析。利用SPSS17.0软件分析两组人群中mtDNA D-loop区多态位点出现频率的差异。结果 1. 与健康对照组相比，肾透明细胞癌患者的年龄、性别分布差异无统计学意义（P>0.05）。在健康对照组和肾透明细胞癌组两组人群中的mtDNA D-loop 区共发现143个多态性位点。2与健康对照组相比，肾透明细胞癌组患者的mtDNA D-loop区的262T，16293G出现的频率明显增高，16298C，16319A出现的频率下降，差异均有统计学意义（P<0.05）。结论 肾透明细胞癌的发病与年龄、性别无关，分析mtDNA D-loop 区域多态性有利于确定肾细胞癌的亚型和重新制定临床治疗策略。

关键词: D-loop, 肾透明细胞癌, SNP, 线粒体DNA

Abstract: Objective: To investigate the relationship between single nucleotide polymorphisms in mitochondrial displacement-loop and the risk of renal cell carcinoma. Methods: We selected 59 clear cell renal cell cancer patients who received nephrectomy in the Department of urinary surgery at the Fourth Hospital of Hebei University between 2002 and 2007. We also collected 68 healthy female controls. Total DNA was extracted using a Wizard Genomic DNA extraction kit (Promega, Madison, WI, USA) and stored at -20℃. PCR was performed according to the protocol of PCR Master Mix Kit (Promega) and purified prior to sequencing. Cycle sequencing was performed with the Dye Terminator Cycle Sequencing Ready Reaction Kit (Applied Biosystem, Foster City, CA, USA) and the products were then separated on the ABIPRISM Genetic Analyzer 3100 (Applied Biosystem). Polymorphisms were confirmed by repeated analyses from both strands. Results: SNPs were detected in 143 sites of the 982-bp mitochondria D-Loop region from blood samples of the ccRCC patients and the healthy controls. No statistical reference exist for distribution frequency of each SNP referring to age and sex. A statistically significant increase in SNP frequency for the 16293G, 262T alleles was observed in clear cell renal cancer patients (p<0.05), whereas the SNP frequency for the 16298C, 319A alleles was decreased in clear cell renal cell cancer patients. Conclusion: analysis of genetic polymorphisms in the D-loop may help to identify patient subgroups at a higher risk for developing RCC, thereby helping to refine therapeutic decisions for these patients.

Key words: D-loop, clear cell renal cell carcinoma, SNP, mtDNA