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目的检测人胰腺癌细胞株PANC-1的K-ras基因点突变方式,明确干扰Ras基因靶点的碱基序列。方法针对K-ras基因第十二位点密码子点突变最常见方式CAT,CGT,GTT设计顺序特异性引物(SSP),对人胰腺癌细胞株PANC1进行聚合酶链反应(PCR),扩增产物用聚丙烯酰胺凝胶电泳判定该细胞株有无点突变及突变方式。结果人胰腺癌细胞株PANC-1存在K-ras基因的点突变,其突变方式为CAT。结论 PCR-SSP法快速简便,特异性高,结果准确,为应用RNAi技术研究胰腺癌的基因治疗奠定了基础。
Objective To detect the point mutation of K-ras gene in human pancreatic cancer cell line PANC-1 and to clear the base sequence of Ras gene target. Methods The sequence-specific primers (SSPs) designed for the codon point mutations of the 12th site of K-ras gene were cloned by CAT, CGT and GTT. The human pancreatic cancer cell line PANC1 was amplified by polymerase chain reaction (PCR) The product was determined by polyacrylamide gel electrophoresis with no point mutation and mutation. Results The point mutation of K-ras gene was found in human pancreatic cancer cell line PANC-1. The mutation was CAT. Conclusion The PCR-SSP method is rapid, simple, accurate and accurate. It laid the foundation for gene therapy of pancreatic cancer using RNAi technique.