高表达人源Her2乳腺癌小鼠模型的建立及鉴定

发布时间：2018-01-03 11:35

本文关键词：高表达人源Her2乳腺癌小鼠模型的建立及鉴定　出处：《第二军医大学学报》2017年04期 　论文类型：期刊论文

更多相关文章： 乳腺肿瘤 转基因小鼠 动物模型 Her

【摘要】：目的构建并鉴定高表达人源Her2乳腺癌转基因小鼠模型。方法构建pMD18T-MMTV-huHER2-EGFP转基因构件,通过原核显微注射方法将线性化、纯化后的外源质粒pMD18T-MMTV-huHER2-EGFP注射入C57BL/6J小鼠受精卵中,胚胎移植至同期发情的假孕受体母鼠输卵管内,获得子代小鼠。采用PCR鉴定子代小鼠尾部组织基因组DNA,用蛋白质免疫印迹法检测乳腺组织Her2蛋白的表达,通过组织病理学切片观察转基因小鼠乳腺组织的病理学变化。结果经PCR方法检测得到转基因阳性小鼠。蛋白质免疫印迹结果显示,与野生型小鼠相比,F2代阳性小鼠的乳腺组织中Her2蛋白高表达。病理组织切片表明25周龄阳性小鼠的乳腺组织已经出现明显的癌变倾向。结论高表达人源Her2乳腺癌小鼠模型成功建立并能稳定遗传,可自发形成乳腺癌,其生物学特性和病理学改变与人乳腺癌相近,可作为研究乳腺癌发生发展的动物模型。
[Abstract]:Objective to construct and identify the high expression of human breast cancer Her2 transgenic mouse model. Methods we constructed pMD18T-MMTV-huHER2-EGFP transgenic components by pronuclear microinjection method will be linear, the plasmid pMD18T-MMTV-huHER2-EGFP C57BL/6J was injected into mouse zygotes after purification, embryo transfer to pseudopregnant pseudopregnant female fallopian tube of estrus, the offspring were obtained. The genomic DNA PCR identification of offspring mouse tail tissue expression using Western blot analysis of Her2 protein in breast tissue, pathological changes were observed in transgenic mice mammary gland tissues by histological sections. The results detected by PCR from transgenic mice. Western blot results showed that, compared with wild-type mice, Her2 protein the expression of F2 positive mouse mammary tissues. Pathological sections showed that breast group 25 week old mice positive fabric There has been a noticeable tendency of canceration. Conclusion the highly expressed mouse model of human Her2 breast cancer is successfully established and stable, which can spontaneously form breast cancer. Its biological characteristics and pathological changes are similar to those of human breast cancer. It can be used as an animal model to study the occurrence and development of breast cancer.

【作者单位】：浙江嘉兴学院医学院医学实验中心;浙江嘉兴学院医学院药学系;第二军医大学海军医学系放射医学教研室;
【基金】：浙江省科技厅项目(2014C37018,2017C37118,2016C37097)~~
【分类号】：R737.9;R-332
【正文快照】： 乳腺癌是女性恶性肿瘤死亡威胁的第二大元凶,需耗费巨额的人力、物力、财力用于乳腺癌治疗的研究[1]。研发一种新的乳腺癌药物,平均要花费6.1亿美元,还要进行为期37个月的动物实验[2]。因此,研究者们前期会应用乳腺癌动物模型来筛选有效的治疗方法,以期为后续的临床试验提供强

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