大黄素和叠氮胸苷对Egr-1 siRNA转染的KG-1a细胞的增殖和凋亡的影响  

Effects of the Combination of Emodin and 3'-Azido-3'-Deoxythymidine on Proliferation and Apoptosis in leukemia KG-1a cells Transfected with Egr-1 siRNA

作　　者：原凌燕[1] 陈彻[1] 楚惠媛[1] 刘玉[1] 

机构地区：[1]甘肃中医学院医学技术学院,甘肃兰州730000

出　　处：《中国实验血液学杂志》2015年第2期386-391,共6页Journal of Experimental Hematology

基　　金：甘肃省财政厅基本科研业务费专项资金(甘政财2010-176)

摘　　要：目的:本研究旨在观察Egr-1基因KG-1a细胞转染后,大黄素联合AZT对KG-1a细胞增殖的影响及其作用机理。方法:采用电转法瞬时转染Egr-1 siRNA,细胞分为空白对照、非特异对照(转染非特异序列)和目的 siRNA(转染Egr-1 siRNA)3组。流式细胞术测定转染效率,MTT法检测各组细胞增殖率,实时荧光定量PCR检测Egr-1基因的表达水平。各组细胞经大黄素10μmol/L或/和AZT 3200、1600μmol/L作用后,应用MTT法检测细胞增殖抑制率,流式细胞术检测细胞凋亡率。结果:电转染Egr-1 siRNA转染效率可达59.21%以上,目的 siRNA组与空白对照和非特异性对照组相比,细胞增殖明显增加,Egr-1 mRNA的表达水平显著下降(P<0.001);大黄素和AZT联合对空白对照和非特异性对照组细胞的增殖抑制均呈现协同作用,联合作用指数(CI)均小于1,而对目的siRNA组细胞的CI大于1,增殖抑制作用与单用AZT相似(P>0.05)。目的 siRNA组细胞凋亡率较空白对照降低(P<0.001)。结论:大黄素联合AZT协同抑制KG-1a细胞增殖和促进凋亡的作用与Egr-1基因有关。Objective： This study was aimed to investigate the effects of emodin combined with 3＇-azido-3＇- deoxythymidine （AZT） on proliferation and apoptosis of leukemia cell line KG-la cells and its mechanism. Methods： KG-la cells were transfected with Egr-1 siRNA by electroporation and divided into blank control （KG-la）, nonspecific control （KG-la/NC） and Egr-1 siRNA （KG-la/siRNA） groups. Transfection efficiency was tested through fluorescence microscopy and flow cytometry and the transfection effect was detected by using qPCR. The cell proliferation rate was detected with MTT method. After the cells were treated with 10 vmol/L of emodin, 3200 or 1600 ixmol/L of AZT and their combinations, the proliferation inhibition rates and the apoptosis rates of cells in 3 groups were detected with MTT method and FCM, respectively. Results： The transfection efficiency of Egr-1 siRNA was found to reach more than 59.21% ; as compared with blank control（KG-la） and nonspectic control （ KG-la/NC）, the cell proliferation in Egr-1 siRNA group significantly reduced （ P 〈 0. 01 ）. The combination of emodin and AZT had considerable synergistic inhibitory effects on proliferation of normal KG-1 a cells and nonspecific control （KG-la NC） cells, but the synergistic effects disappeared after Egr-1 gene silencing. Conclusion： The effects of the combination of emodin and AZT on troliferation and anoDtosis mav be related with Egr-1.

关 键 词：EGR-1基因 KG-1a细胞 大黄素 叠氮胸苷 

分 类 号：R733.71[医药卫生—肿瘤]