Aiolos基因过表达对白血病细胞生物学特性及化疗敏感性的影响

Influence of over-expressed Aiolos on cell biological behaviors and chemotherapy sensitivity of leukemic cells

目的研究Aiolos基因过表达对白血病细胞周期、凋亡及化疗敏感性的影响并探讨其机制。方法将Jurkat细胞分为Aiolos过表达组、转染GFP组和未转染组,构建Aiolos基因慢病毒载体,感染Jurkat细胞并检测3组细胞的细胞周期、凋亡、相关基因表达及对依托泊苷敏感性。结果 Aiolos转染Jurkat细胞4 d后,Aiolos过表达组和转染GFP组的GFP表达量达80%以上,且Aiolos过表达组的Aiolos蛋白表达较转染GFP组和未转染组明显上升(P<0.05)。Aiolos过表达组细胞凋亡比例较转染GFP组和未转染组明显增加(P<0.05),细胞周期由G0/G1进入S期比例明显减少(P<0.05)。周期相关基因Cyclin D3和Skp2表达下调(P<0.05),细胞周期蛋白依赖性激酶抑制因子P21和P27表达上调(P<0.05);凋亡相关基因Bax表达无明显变化,Bcl-2表达下调(P<0.05),Bax/Bcl-2比例增加(P<0.05)。Aiolos过表达组细胞在依托泊苷浓度40μg/mL作用36、48 h后,细胞存活百分率较转染GFP组和未转染组均明显降低(P<0.05)。结论在Jurkat细胞系中介导Aiolos基因过表达能够促进细胞凋亡、抑制细胞周期并提高细胞对化疗药物的敏感性。

Objective To evaluate the impact of Aiolos gene over-expression on the cell cycle, apoptosis, chemotherapy sensitivity of leukemia cell lines and the potential mechanism. Methods The Jurkat cells were divided into the Aiolos overexpression group, GFP transfected group and non-transfection group. Aiolos-overexpressed Jurkat ceils were constructed by lentiviral transduction. Then the cell cycle, apoptosis, chemotherapy sensitization and expression of related genes were detected. Results Expression of Aiolos was up-regulated 4 days after transfection. Over-expression of Aiolos promoted cell apoptosis and arrested cell cycle into S phase. In addition, over-expression of Aiolos led to up-regulation of p21, p27 （ P 〈 0.05 ） and down-regulation of cyclin D3, Skp2 and Bcl-2 （ P 〈 0.05 ）. No changes were detected on Bax. However, there was an increase in the Bax to Bcl-2 ratio. Furthermore, the cell survival rates exposed to 40 Ixg/mL etoposide were reduced in the Aiolos overexpression group compared with those in the control groups at 36 h and 48 h （ P 〈 0.05 ）. Conclusion Recombinant lentiviral vectors of Aiolos gene overexpression promote cell apoptosis, arrest cell cycle and enhance etoposide cytotoxity in Jurkat cells.