脂质体法pcDNA3-ELAC2质粒转染乳腺癌细胞MCF-7和MDA-MB-231以及前列腺癌细胞Du-145

Transfection of lipo-mediated pcDNA3-ELAC2 into breast cancer cells MCF-7and MDA-MB-231 and prostate cancer cells Du-145

目的:构建稳定、高效表达ELAC2基因的乳腺癌细胞模型MCF-7/ELAC2、MDA-MB-231/ELAC2,以及前列腺癌细胞模型Du-145/ELAC2,为探索ELAC2基因的生物学功能奠定实验基础。方法:将构建pcDNA3-ELAC2的质粒采用脂质体法转染乳腺癌细胞和前列腺癌细胞,用RT-PCR和WesternBlot分别检测ELAC2基因在细胞中的mRNA表达和蛋白表达。结果:pcDNA3-ELAC2质粒构建成功并顺利导入乳腺癌细胞和前列腺癌细胞中,且检测到ELAC2基因的mRNA和蛋白高表达。结论:构建的pcDNA3-ELAC2重组质粒能够在转染的乳腺癌细胞和前列腺癌细胞中稳定、持续和高效地表达,为进一步研究ELAC2基因的生物学功能提供了理想的实验模型。

Objective： To construct the transgenic cell model MCF-7/ELAC2, MDA-MB-231/ ELAC2 and Du-145/ ELAC2 that expresses stable high amounts of ELAC2, which set up experimental foundation to study biological function of ELAC2 gene. Methods. The recombinant plasmid pcDNA3-- ELAC2 was transfected into the breast cancer cell lines and prostate cancer cell lines by using lipofectamine transfection reagent. Then the ELAC2 mRNA and protein were detected by RT-PCR and Western blot, respectively. Results： pcDNA3-ELAC2 was successfully constructed and transfected into breast cancer cells and prostate cancer cells. RT-PCR and Western blot showed that the transfected cells had high ELAC2 expression at both mRNA and protein level. Conclusion. Recombinant plasmid of pcDNA3-ELAC2could have stable, cancer cells, which ELAC2 gene. sustainable and efficient expression in provide an ideal experimental model the transfected breast cancer cells and prostate for further study of the biological function of ELAC2 gene.