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目的研究从厦门同安肝癌高发区所建的裸鼠移植瘤(HHC4、HHC15)是否存在p53基因突变。方法从两移瘤提取的总DNA用分子生物学方法检测并测序,如PCR方法扩增p53基因部分第7外显子,地高辛标记DNA探针斑点杂交法,DNA限制性酶切片段长度多态分析和激光荧光DNA测序等。结果HHC4细胞DNA有p53基因第250密码子(C→A)的突变;HHC15有p53基因第249密码子(G→T)的突变。结论HHC4、HHC15p53基因突变是肝癌发生的基础,与移植瘤来源地区AFB1的严重污染可能有关系。
Objective To investigate whether p53 gene mutation exists in transplanted tumors (HHC4, HHC15) in nude mice constructed from high-risk areas of Tongan liver cancer in Xiamen. Methods The total DNA extracted from the two tumors was detected by molecular biological methods and sequenced, such as PCR amplification of the exon 7 part of the p53 gene, digoxigenin-labeled DNA probe dot blot hybridization, DNA restriction fragment length Polymorphic analysis and laser fluorescence DNA sequencing. RESULTS: DNA of HHC4 cells had a mutation at codon 250 of the p53 gene (C→A); HHC15 had a mutation at position 249 of the p53 gene (G→T). Conclusion The HHC4 and HHC15p53 gene mutations are the basis of HCC and may be related to the severe contamination of AFB1 in the area of transplanted tumors.