阳性对照PCR-SSCP法检测肝癌中特异性p53基因点突变

Detection of the Tumor-Specific p53 Codon 249 Mutation in Human Hepatocellular Carcinomas by Positive -Control PCR-SSCP

用人肝癌细胞系PLC/PRF/5 DNA作阳性对照,通过聚合酶链式反应—单链构象多态分析(PCR-SSCP分析),检测五种人肝癌细胞系及6例原发性肝癌组织中p53基因点突变。在被检标本中未检出PLC/PRF/5中存在的肝癌特异性p53基因点突变,即249位密码子第3碱基G→T颠换。因此,这种特异性p53基因点突变并不是肝癌中的普遍现象。

The tumor-specific p53 codon 249 mutation had been frequently observed in human hepatocellular carcinomas(HCCs). DNA samples from 5 HCC cell lines and 6 HCC tissues were subjected to by the polymerase chain reaction-single strand conformation polymorphism analysis (PCR-SSCP analysis) to detect the hotspot mutation. DNA from PLC/PRF/5 cells in which the mutation, a G→T transversion at the third base of codon 249, had been confirmed was selected as the positive control. No singlestrand patterns showing the PLC/PRF/5 specific mobility shifts were detected. This suggested that the codon 249 mutation lacked in the DNA samples examined. The result showed the eodon 249 mutation was not a common event in hepatocarcinogenesis. The present study was significant for molecular etiology of HCC and for rapid detection of known mutations.