HER2基因转染MCF-7细胞及生物学特性观察

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

目的:建立共表达HER2(HumanEpidermalGrowthFactorReceptor2)基因和ER(EstrogenReceptor)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、Westernblot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。

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.