稳定表达PTEN抑癌基因的炎性乳腺癌细胞系MDA-MB-468的建立和鉴定

目的建立稳定表达外源性PTEN(phosphataseandtensinhomologdeletedfromchromosometen)基因的人炎性乳腺癌MDA MB 468细胞系。方法用脂质体lipofectamine2000转染法,将野生型PTEN基因导入高转移性炎性乳腺癌细胞系MDA MB 468,经G418筛选抗性克隆,扩大培养后,应用RT PCR、免疫组化方法及Westernblot分析目的基因及其蛋白表达。结果野生型PTEN基因成功地导入炎性乳腺癌细胞系MDA MB 468,转染细胞稳定表达PTEN蛋白,PTEN蛋白分布于细胞质。结论MDA MB 468炎性乳腺癌细胞株能稳定表达外源性PTEN基因。

The establishment of stable transfection of human breast cancer cell line MDA-MB-468 with exogenous PTEN gene

Objective To investigate exogenous PTEN gene transfected human breast cancer cell line MDA MB 468. MethodsUsing the lipofectamine 2000 transfection technique,wild type PTEN gene was transducted into an in vitro cultured highly metastatic breast cancer cell line MDA MB 468.After transfection, the cells were selected by G418.Then resistant clones were chosen and expanded in DMEM culture medium. RT PCR, immunohistochemical method and western blot were used to determine the expression of target genes. ResultsAn anti G418 cell clone was established and expanded in culture. The transfected PTEN gene MDA MB 468 cells showed expression of PTEN mRNA and PTEN protein. ConclusionHuman breast cancer cell line MDA MB 468 established in this study expresses consistently exogenous PTEN genes.