Shi Xianzhe, Xu Guowang, Zhao Chunxia, Ma Jianmei, Zhang Yan, Lv Shen, Yang Qing
National Chromatographic R & A Center, Dalian Institute of Chemical Physics, the Chinese Academy of Sciences, Dalian, China.
Electrophoresis. 2003 Jul;24(14):2316-21. doi: 10.1002/elps.200305444.
Mutation of hMLH1 gene plays an important role in human tumorigenesis. A highly sensitive single-strand conformation polymorphism (SSCP) method for detection of the T1151A mutation in exon 12 of the hMLH1 gene was for the first time developed employing laser-induced fluorescence capillary electrophoresis (LIF-CE). Effects of the concentration of linear polyacrylamide solution, running temperature, running voltage and the addition of glycerol on SSCP analysis were investigated, and the optimum separation conditions were defined. Thirty colorectal cancer patients and eight lung cancer patients were screened and the T1151A mutation was found in four of them. Based on CE-sequencing the mutation was further confirmed. To our knowledge, this is for the first time that the T1151A mutation is found in lung cancer. Our method is simple, rapid, and highly sensitive and is well suited to the analysis of large numbers of clinical samples.
hMLH1基因的突变在人类肿瘤发生过程中起着重要作用。首次开发了一种利用激光诱导荧光毛细管电泳（LIF-CE）检测hMLH1基因第12外显子T1151A突变的高度灵敏的单链构象多态性（SSCP）方法。研究了线性聚丙烯酰胺溶液浓度、运行温度、运行电压以及甘油添加量对SSCP分析的影响，并确定了最佳分离条件。对30例结直肠癌患者和8例肺癌患者进行筛查，其中4例发现T1151A突变。通过CE测序进一步证实了该突变。据我们所知，这是首次在肺癌中发现T1151A突变。我们的方法简单、快速且高度灵敏，非常适合分析大量临床样本。
