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目的确定α粒子诱发的恶性转化大鼠气管上皮(rattrachealepithelial,RTE)细胞中是否存在p53基因突变。方法根据大鼠p53cDNA序列,合成4对引物分别覆盖大鼠p53基因外显子5,6,7和8,利用聚合酶链反应-单链构象多态方法分析转化细胞中的p53突变情况。结果在转化细胞,p53基因外显子8的单链条带与正常细胞相比出现泳动变位,经测序证实为265密码子G→C突变,使其编码氨基酸由精氨酸→脯氨酸。结论在α粒子诱发的恶性转化RTE细胞中存在p53基因突变,提示p53基因变异在辐射致癌过程中可能起重要作用
OBJECTIVE: To determine whether there is p53 mutation in rat α-particle-induced malignant transformed rat tracheal epithelial cells (RTE). Methods According to the rat p53 cDNA sequence, 4 pairs of primers were synthesized to cover the exons 5, 6, 7 and 8 of p53 gene respectively. The mutation of p53 in transformed cells was analyzed by polymerase chain reaction - single strand conformation polymorphism. Results In the transformed cells, the single-stranded band of exon 8 of p53 gene appeared to migrate compared with that of normal cells. The sequence of G → C mutation at codon 265 confirmed that the encoded amino acid consists of arginine → proline . Conclusion The mutation of p53 gene exists in α-particle-induced malignant transformed RTE cells, suggesting that p53 gene mutation may play an important role in radiation-induced carcinogenesis