医学分子生物学杂志2011，Vol.8Issue(6)：489-493,5.DOI:10.3870/j.issn.1672-8009.2011.06.005
Akt3稳定表达细胞株的建立及其对MDA-MB-231细胞增殖的影响
Effect of Stable Expression of Akt3 on Proliferation of MDA-MB-231 Cells
摘要
Abstract
Objective To construct eukaryotic expression vector pEGFP-N2-Akt3, to establish a cell clone stably expressing Akt3 and to study the effect of Akt3 on proliferation of breast cancer cells. Methods Total RNA was extracted from fetal brain tissue. Akt3 cDNA was amplified by RT-PCR, and inserted into pEGFP-N2 vector to construct pEGFP-N2-Akt3. The recombinant plasmid was then transfected into MDA-MB-231 cells by lipofectamine. The stable transfectants of MDA-MB-231 cells were selected with G418. Western Blot was used to detect AKT3 expression level. MTT assay were used to evaluate the effects of Akt3 on MDA-MB-231 cell proliferation. Results The recombinant expression vector pEGFP-N2-Akt3 was successfully constructed. The expression of AKT3-EGFP fusion protein was only detected in MDA-MB-231 cells transfected with the Akt3 expression vector, but not in empty vector transfected or untransfected MDA-MB-231 cells. MTT assay showed that the proliferation of MDA-MB-231 cells with Akt3 stable expression was significantly higher than that of control cells ( P <0. 01 ) . Conclusion Akt3 can promote MDA-MB-231 cell proliferation.关键词
蛋白激酶Bγ/基因克隆/基因转染/基因表达/四唑盐比色测定Key words
AKT3 / gene cloning/ gene transfection/ gene expression/ MTT assay分类
生物科学引用本文复制引用
韩俊永,黄俏佳..Akt3稳定表达细胞株的建立及其对MDA-MB-231细胞增殖的影响[J].医学分子生物学杂志,2011,8(6):489-493,5.基金项目
南京军区医药卫生科研基金(No.08MA100),福建省自然科学基金(No.2009J01181) （No.08MA100）
