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目的研究运城市中心医院收集的71例结直肠癌患者外周血的K-ras基因12位密码子的突变情况及突变类型。方法采用RFLP-/PCR套式技术及酶切法筛查12位密码子突变情况,并通过pMD-18 T载体构建测序分析突变类型。结果 71例结直肠癌患者中,7例腺癌二级患者K-ras基因12位密码子发生突变,其中1例同时发生16位密码子突变,突变率为11%。测序共发现两类突变类型:错义突变(12位:GGT—AGT、GAT;16位:AAG—AGG)和移码突变(GGT—GT)。结论外周血K-ras基因12位密码子PCR突变筛查可以作为结直肠癌早期诊断的一项辅助方法。
Objective To investigate the mutation and mutation of K-ras gene codon 12 in peripheral blood of 71 cases of colorectal cancer collected from Yuncheng Central Hospital. Methods RFLP- / PCR kit and restriction enzyme digestion were used to screen the 12-codon mutations. The mutation types were analyzed by pMD-18 T vector. Results Among the 71 patients with colorectal cancer, seven of the two patients with adenocarcinoma had K-ras gene mutations at codon 12, of which 16 had a codon 16 mutation at the same time. The mutation rate was 11%. Two types of mutations were identified: missense mutation (GGT-AGT, GAT; position 16: AAG-AGG) and frameshift mutation (GGT-GT). Conclusion The screening of mutations in K-ras gene at codon 12 in peripheral blood can be used as an adjunct to early diagnosis of colorectal cancer.