p53/LKB1双缺失子宫内膜癌小鼠模型建立与生物学鉴定  

Establishment of the endometrial cancer mouse model with dual loss of p53/LKB1 and its biological characteristics study

作　　者：郭慧[1,2] 盛修贵[2] 韩晓运[2] 张璐[1,2] 周春晓[3] 

机构地区：[1]济南大学.山东省医学科学院医学与生命科学学院,山东济南250200 [2]山东大学附属山东省肿瘤医院妇瘤科,山东济南250117 [3]北卡罗来纳大学教堂山分校妇瘤科

出　　处：《中华肿瘤防治杂志》2016年第9期563-567,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金重点项目(30930038);国家自然科学基金(81302292)

摘　　要：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 Recenthy,the accidence of endometrial cancer has increeased. The loss of LKB1 gene has been shown to be one ipportant factor driving endometrial cancer. To establish a genetically engineered mouse model of endometrial cancer induced by dual loss of tomor suppressor p53 and LKB1 for studying endometrial cancer progression. METHODS The genetype of mouse model was detected by PCR. We utilized a unique endometrial cancer mouse model that specifically deleted p53 and LKB1 tumor suppressor gene via microinjection of Ad-CMV-Cre adnovirus（AdCre） into one side of mouse uterus. The histological morphon of uterus tumor was analyzed by H＆E staining. RESULTS The transgenic mouse model of p53^loxp/loxpLKB1^loxp/loxp was achieved. The target gene was verified to contain loxp sites. The earliest atypical endometrial hyperplasia appeared in 2 weeks after AdCre injection. At 4 weeks, around 25% endometrial adenocarcinoma developed in the affected uterus,while the uninjected uterus remains normal. At 12 weeks, the incidence of mouse with endometrial adenocarcinoma was 100%. The longest survival time of the mice with tumor was 34 weeks.At 20 weeks after AdCre injection, about 33% of LKB1^loxp/loxp or p53^loxp/loxpLKB1^loxp/loxp mice displayed endometrial adeno- careinomar and their average survival time was 52 weeks. CONCLUSIONS By taking p53 and LKB1 conditional knockout in transgenic mouse uterus under AdCre injection, we successfully generated an endometrial adenocarcimoma mouse mod el. This method can shorten the time of inducement and increase the incidence of tumor. Its histopathological characteris- tics met the criterion of endometrial adenocarcinoma process, and it is an ideal mouse model used for studying the occur- rence and development of endometrial cancer.

关 键 词：子宫内膜癌 小鼠 P53 LKB1 转基因 

分 类 号：R737.33[医药卫生—肿瘤]