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中国癌症防治杂志

中国癌症防治杂志 ›› 2013, Vol. 5 ›› Issue (1): 35-38.doi: 10.3969/j.issn.1674-5671.2013.01.09

• 基础研究 • 上一篇    下一篇

人肝癌SMMC-7721细胞中POLD1基因CDS突变的检测

  

  • 出版日期:2013-03-25 发布日期:2013-04-07
  • 通讯作者: 徐恒 E-mail:dnareplication@163.com
  • 基金资助:

    国家自然科学基金资助项目（30950028）；广西自然科学基金资助项目（2010GXNSFC013017）

Mutational analysis of the protein-coding sequence of the POLD1 gene in human hepato-cellular carcinoma cell line SMMC-7721

  • Online:2013-03-25 Published:2013-04-07

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摘要： 目的 检测人肝癌细胞株SMMC-7721中POLD1基因CDS的突变情况，以期从DNA复制最关键酶的编码基因POLD1的突变为干预细胞恶性增殖提供新的思路。方法 设计人POLD1基因CDS特异性引物，以人肝癌细胞株SMMC-7721总RNA为模板进行RT-PCR扩增，将扩增产物纯化后加入“腺嘌呤”并与T载体连接，将连接产物进行测序，与数据库中的序列进行比对。结果 成功扩增了人肝癌细胞中POLD1基因的CDS片段。SMMC-7721细胞中POLD1基因的CDS存在11个碱基替换，1个碱基缺失。结论 人肝癌细胞SMMC-7721中POLD1基因的CDS发生了突变，进而导致其编码的氨基酸序列发生改变。

关键词: 肝肿瘤, POLD1, 突变

Abstract:  Objective To analyze mutations in the protein-coding region of the POLD1 gene in the human hepatocarcinoma cell line SMMC-7721；this gene encodes an enzyme critical for DNA replication. Methods Total RNA was isolated from SMMC-7721 cultures，and the POLD1 gene coding region was amplified by RT-PCR，purified and A-tailed， and then cloned into a T-vector. Mutations were detected by sequencing. Results The protein-coding region of the POLD1 gene was successfully amplified and found to contain 12 mutations，11 of which were nucleotide changes and 1 was a nucleotide deletion. Conclusions SMMC-7721 en-codes a mutated POLD1 gene， and the mutations lead to changes in the protein sequence.

Key words:  Liver neoplasms, POLD1, Mutation