色谱

色谱 ›› 2013, Vol. 31 ›› Issue (6): 582-586.DOI: 10.3724/SP.J.1123.2012.12023

• 研究论文 • 上一篇    下一篇

结直肠癌组织中K-ras基因突变的毛细管电泳检测

石冬琴1,2, 王荣1,2, 谢华1, 田薇2, 贾正平1, 郭建魁1   

  1. 1. 全军高原损伤防治重点实验室, 兰州军区兰州总医院临床药理基地, 甘肃 兰州 730050;
    2. 浙江农林大学林业与生物技术学院, 浙江 临安 311300
  • 收稿日期:2012-12-12 修回日期:2013-01-14 出版日期:2013-06-28 发布日期:2013-06-06
  • 通讯作者: 王荣, 贾正平
  • 基金资助:

    国家自然科学基金项目(20775089);全军医药卫生"十一五"科研课题(06MB096).

Determination of K-ras gene mutation in colorectal cancer tissue by capillary electrophoresis

SHI Dongqin1,2, WANG Rong1,2, XIE Hua1, TIAN Wei2, JIA Zhengping1, GUO Jiankui1   

  1. 1. PLA Key Laboratory of the Plateau of the Environmental Damage Control, Base of Clinic Pharmacology, Lanzhou General Hospital, Lanzhou 730050, China;
    2. Forest and Biotechnology College, Zhejiang Agriculture and Forest University, Lin’an 311300, China
  • Received:2012-12-12 Revised:2013-01-14 Online:2013-06-28 Published:2013-06-06
  • Contact: 10.3724/SP.J.1123.2012.12023

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摘要：

通过对PCR扩增的76例结直肠癌组织及癌旁正常组织DNA基因组共152个样本纯化变性后,采用毛细管电泳-激光诱导荧光检测(CE-LIF)结合单链构象多态性(SSCP)分析方法检测了人结直肠癌组织及癌旁正常组织中K-ras基因第12/13位密码子突变。所检测的76例结直肠癌患者中有30例患者存在基因突变,并对异常片段进行测序验证,测序证实以碱基G→A点突变为主。结果表明所建立的CE-LIF技术结合SSCP分析检测K-ras基因突变的方法高效、快速、灵敏、准确,适合于临床上大样本结直肠癌中K-ras基因突变分析,对选择抗结直肠癌药物有一定的指导作用。

关键词: K-ras基因, 单链构象多态性, 基因突变, 结直肠癌组织, 毛细管电泳

Abstract:

The codons 12/13 of K-ras genomic DNA from 76 colorectal cancer tissues and normal tissues were amplified by PCR. The amplified 152 samples were purified, denatured, and then detected using capillary electrophoresis (CE)-laser induced fluorescence (LIF) with single-strand conformation polymorphism (SSCP). The abnormal samples were further confirmed by direct-sequencing. The 30 patients of the 76 colorectal cancer patients were found gene mutation, of which the results of base G→A point mutation were confirmed by the gene sequence. To detect K-ras gene mutation that has important role in clinical forecast, diagnosis, therapy and prognosis, the CE-LIF with SSCP was applied for detecting K-ras gene. It becomes a promising tool to analyse K-ras gene mutation in the clinical diagnosis and therapy of colorectal cancer tissue.

Key words: capillary electrophoresis (CE), colorectal cancer tissue, gene mutation, K-ras gene, single-strand conformation polymorphism

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