人肝癌裸鼠移植瘤株p53基因突变及测序研究

Studies of p53 Gene Mutation in the Xenografts of Human Hepatocellular Carcinoma in Nude Mice

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 presented in the xenografts(HHC 4,HHC 15 ) of nude mice set up in Tong’an Area Xiamen with high mortality of HHC.\ Methods\ Total DNA extracted from two xenografts were analysed by molecular biological methods,such as PCR amplification of a fragement of p53 extron 7,dot blot filter hybridization with digoxigeninlabeled DNA probe,restriction fragments length polymorphisms and DNA sequencing.\ Results\ The mutation at 250 coden(C→A) in p53 was determined in DNA of HHC 4 xenograft and at 249 coden(G→T) in DNA of HHC 15 .\ Conclusions\ It is reasonable that p53 gene mutation is responsible for the carcinogenesis of HHC and may be a result of the high pollution of AFB 1.