Shi Dongqin, Wang Rong, Xie Hua, Tian Wei, Jia Zhengping, Guo Jiankui
PLA Key Laboratory of the Plateau of the Environmental Damage Control, Base of Clinic Pharmacology, Lanzhou General Hospital, Lanzhou 730050, China.
Se Pu. 2013 Jun;31(6):582-6. doi: 10.3724/sp.j.1123.2012.12023.
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.
采用聚合酶链反应（PCR）扩增76例结直肠癌组织及正常组织中K-ras基因组DNA的第12/13密码子。将扩增后的152个样本进行纯化、变性，然后采用毛细管电泳（CE）-激光诱导荧光（LIF）结合单链构象多态性（SSCP）进行检测。对异常样本进一步行直接测序确认。76例结直肠癌患者中有30例发现基因突变，其中碱基G→A点突变结果经基因序列确认。为检测在临床预测、诊断、治疗及预后中具有重要作用的K-ras基因突变，应用CE-LIF结合SSCP检测K-ras基因。它成为结直肠癌组织临床诊断及治疗中分析K-ras基因突变的一种有前景的工具。
