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目的近年来,子宫内膜癌发病率逐年上升,LKB1基因缺失是引发子宫内膜癌的一个重要原因。建立由p53loxp/loxpLKB1loxp/loxp双缺失致子宫内膜腺癌的小鼠模型,为研究子宫内膜肿瘤的发生发展提供动物模型。方法选取6周龄p53和LKB1小鼠雌雄各2只。PCR鉴定拟诱导模型小鼠的基因型。显微镜下将Ad5-CMV-Cre(AdCre)腺病毒注射到p53loxp/loxpLKB1loxp/loxp基因型小鼠的一侧子宫内,诱导子宫内膜肿瘤形成。HE切片染色确定子宫肿瘤的类型。结果成功获得基因型为p53loxp/loxpLKB1loxp/loxp的转基因小鼠。经PCR鉴定目的基因含有loxp位点。该小鼠子宫AdCre腺病毒注射后最早第2周出现不典型增生,4周时约25%HE切片染色诊断为子宫内膜腺癌,而未注射侧子宫正常;12周左右出瘤率为100%,荷瘤生存时间≤34周。LKB1loxp/loxp与p53loxp/-LKB1loxp/loxp基因型小鼠注射侧子宫20周出瘤率约为33%,荷瘤生存时间平均为52周。结论小鼠模型病理学示,腺体不规则,腺管排列拥挤、紊乱,异型性细胞增多,可见核分裂像,符合子宫内膜腺癌特征。通过显微注射AdCre腺病毒于小鼠子宫内,成功敲除p53及LKB1基因,建立子宫内膜腺癌转基因小鼠模型,该模型成瘤时间短,出瘤率高,其病理表现与人类子宫内膜腺癌类同,是研究子宫内膜癌发生发展的理想小鼠模型。
Objective In recent years, the incidence of endometrial cancer increased year by year, LKB1 gene deletion is an important cause of endometrial cancer. To establish a mouse model of p53loxp / loxpLKB1loxp / loxp double deletion endometrial adenocarcinoma and to provide an animal model for studying the occurrence and development of endometrial tumors. Methods Two female and one male, 6 weeks old, p53 and LKB1 mice were selected. PCR to identify the model of induced mice genotype. Ad5-CMV-Cre (AdCre) adenovirus was injected microscopically into the uterus of one side of a p53loxp / loxpLKB1loxp / loxp genotype mouse to induce endometrial neoplasia. HE slice staining to determine the type of uterine tumor. Results Transgenic mice with genotype p53loxp / loxpLKB1loxp / loxp were successfully obtained. The target gene was identified by PCR containing loxp sites. The mouse uterus AdCre adenovirus injection of the first 2 weeks after the onset of atypical hyperplasia, 4 weeks, about 25% of HE slice staining diagnosis of endometrial adenocarcinoma, uterus without injection side of the normal; about 12 weeks the tumor was 100 %, Tumor-bearing survival time ≤34 weeks. The rate of tumor formation in the LKB1loxp / loxp and p53loxp / -LKB1loxp / loxp mice at 20 weeks after injection was about 33% and the average tumor-bearing survival time was 52 weeks. Conclusion The mouse model showed pathology, irregular glands, crowded and disorganized ductal glands, increased atypia cells, and mitotic figures. It was consistent with the characteristics of endometrial adenocarcinoma. By microinjection of AdCre adenovirus in mouse uterus, successfully knock-out p53 and LKB1 gene, the establishment of endometrial adenocarcinoma transgenic mouse model, the tumor formation time is short, high rate of tumor, the pathological manifestations of human uterus Endometrial adenocarcinoma similar, is to study the development of endometrial cancer ideal mouse model.