高表达人源Her2乳腺癌小鼠模型的建立及鉴定

Establishment and identification of transgenic breast cancer mouse model with Her2 overexpression

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。

Objective To establish a transgenic breast cancer mouse model with human Her2 overexpression and to identify the mouse model. Methods The purified exogenous plasmid pMD18T-MMTV-huHER2-EGFP was constructed and injected into C57BL/6J mouse fertilized eggs by prokaryotic microinjection method. Then the embryos were transplanted into the fallopian tubes of pseudopregnant mice in estrus simultaneously, and the offspring mice were obtained. The genomic DNA of offspring mice was identified by PCR method. The expression of Her2 protein in the breast tissue of offspring mice was detected by Western blotting analysis. The pathological changes of breast tissue in transgenic mice were observed by histopathological section. Results Her2 transgenic mice were detected by PCR method. Western blotting analysis showed that, compared with wild-type mice, the expression of Her2 protein in the breast tissue of F2 generation positive mice was significantly higher. Histopathological sections showed that the breast tissue of 1;2 generation positive mice of 25 weeks old had an obvious carcinogenic tendency. Conclusion The transgenic breast cancer mouse model with human Her2 overexpression has been successfully established, with stable genetic and spontaneous formation of breast cancer. The biological characteristics and pathological changes of the model are similar to human breast cancer, indicating that it can be used as an animal model to study the development and progression of breast cancer.