表达人HER2/neu及荧光素酶的小鼠乳腺癌模型的建立

Establishment of a mouse mammary cancer model stably expressing human HER2/neu gene and luciferase gene

目的建立表达HER2/neu表皮生长因子受体及萤火虫荧光素酶基因的小鼠乳腺癌细胞模型。方法将重组人HER2/neu原癌基因真核表达载体与荧光素酶报告基因依次转染小鼠乳腺癌4T1细胞系,用G418与潮霉素加压筛选阳性克隆,Western印迹及活体成像技术鉴定HER2/neu和荧光素酶在4T1细胞中的表达。在BALB/c小鼠前肢乳腺皮下注射该细胞1.5×106个,活体成像观察肿瘤的生长及转移。结果与结论经过两步转染获得抗G418及潮霉素4T1细胞,Western印迹结果显示,该细胞高表达HER2/neu。流式细胞术检测结果提示几乎所有细胞均有GFP的表达。用该细胞株接种小鼠后具有与野生型4T1细胞相似的成瘤性和转移能力。实验结果提示,我们已建立可表达HER2/neu表皮生长因子受体及荧光素酶基因的小鼠乳腺癌细胞株,为乳腺癌的研究提供一个良好的模型。

Objective To establish a mouse breast cancer model stably expressing human HER2/neu and firefly luciferase gene.Methods 4T1 cells were successively transfected with human HER2/neu and firefly luciferase gene and selected with G418 and hygromycin respectively.The HER2/neu and luciferase protein in selected clones were detected by Western-blot and bioluminescence imaging technology.Finally,the 1.5×106 4T1 cells stably expressing HER2/neu and luciferase gene were transplanted in the mammary fatpad of BALB/c mice.The proliferation and metastasis of the tumor graft were observed and determined with bioluminescence imaging technology.Results and Conclusion After transfection and selection,G418 and hygromycin resistant 4T1 clones were obtained.Western blot analysis showed the transfected 4T1 cells expressed a high level of HER2/neu protein.FACS results showed that almost all the cells from the selected clone expressed GFP protein.Tumor graft experiments showed that the transfected cells had similar in vivo tumorigenesis and metastasis to wild-type 4T1 cells.The results suggest that an HER2/neu-and luciferase-expressing mouse breast cancer model is successfully established.