HER2基因转染MCF-7细胞及生物学特性观察

目的:建立共表达HER2(Human Epidermal Growth Factor Receptor 2)基因和ER(Estrogen Receptor)基因的细胞模型,并观察转染细胞的生物学活性.方法:构建真核表达载体pcDNA3.1-HER2,利用脂质体介导将其转染ER阳性的乳腺癌MCF-7细胞,经G418筛选阳性克隆.用RT-PCR、Western blot及免疫细胞化学等方法检测HER2基因在MCF-7细胞中的稳定表达情况,并通过MTT法和肿瘤细胞侵袭实验,观察转染细胞的生物学活性.结果:在mRNA和蛋白水平证实,转染细胞内有HER2基因的高表达,转染后细胞的增殖能力和侵袭能力明显增强.结论:构建含HER2基因的重组载体,导入乳腺癌MCF-7细胞后,获得生物学活性稳定的HER2基因和ER基因高表达的细胞模型,为进一步研究奠定了基础.

The Biological Characteristics of MCF-7 Cell Line Transfected with HER2 Gene

Objective: To establish a cell line with stable co-expression of HER2 and ER and to study its biological activity. Methods: Recombinant eukaryotic expression vector plasmid pcDNA 3.1-HER2 was constructed. It was transfected into ER expressing MCF-7 cells with lipofectamine. The positive cell clones had been obtained after selected by G418. The MCF-7 cell transfected with HER2-cDNA was determined by RT-PCR, immunocytochemistry and Western blot. The activation of transfected MCF-7 cells was observed by MTT assay and cell invasion assay. Results: The strong expression of HER2 in transfected MCF-7 cells was confirmed at mRNA and protein levels. The capability of proliferation and invasion was obviously enhanced. Conclusion: The recombinant expression vector containing HER2 is successfully constructed. After introduction of the vector into MCF-7 cells, the MCF-7 cell line co-expressing HER2 and ER has also been obtained, which would be helpful for further study on HER2 gene.