转染BAX基因对A549细胞系的凋亡及化疗敏感度的影响
Infl uence of BAX on Apoptosis and Drug Sensitivity of A549 Cells
-
摘要: 目的 探讨BAX转染是否增强人肺癌A549细胞对顺铂致凋亡的敏感度。方法 通过CCK8法检测顺铂对A549细胞体外增殖的影响。应用脂质体介导重组真核表达载体BAX-pcDNA3.1和空载体pcDNA3.1质粒转染至人肺癌A549细胞中，G418筛选阳性细胞。RT-PCR法检测BAX在A549细胞中的表达。将不同浓度的顺铂分别作用于A549、A549/BAX和A549/pcDNA3.1细胞72 h，CCK8法测定各组细胞的存活率，流式细胞术检测细胞周期及细胞凋亡。结果 顺铂能抑制A549细胞体外生长，BAX 和顺铂使A549细胞发生G1/S期细胞阻滞。RT-PCR显示转染BAX基因的A549细胞中BAX表达显著增加，转染BAX基因及加药组细胞凋亡率明显增加。 结论 稳定转染BAX基因明显增强A549细胞对顺铂致凋亡作用的敏感度。Abstract: Objective To investigate the role of BAX in apoptosis and response of human A549 cells to apoptosis induced by Cisplatin(DDP). Methods The proliferation of A549 cells were detected by CCK8 calorimetry after treated with Cisplatin. The plasmid containing BAX cDNA was transfected into A549 cell line by lipofectamine transfection. Expression of BAX was detected by RT-PCR. A549, A549/BAX and A549/pcDNA3 were treated with different concentrations of DDP for 72 h. Survival rate of cells were detected by CCK8. Flow cytometry was used to analyze the apoptosis and cell cycle of A549 cells. Results Cisplatin inhibited the proliferation of A549 cells in vitro. BAX and cisplatin retarded A549 cell at G1/S stage. RT-PCR result showed transfection of BAX vector increased BAX mRNA level, and apoptosis of BAX transfection group and cisplatin treatment group were signifi cantly higher than that in control group. Conclusion Stable transfection of BAX signifi cantly enhances the sensitivity of A549 cells to apoptosis induced by Cisplatin.
下载:
