全反式维甲酸联合猪胆酸钠对K562和Kasumi-1细胞系生物活性抑制作用

本研究旨在探索全反式维甲酸(all-trans retinoic acid,ATRA)联合猪胆酸钠(SBA-Na)对人白血病K562细胞系及Kasumi-1细胞系生物学活性的影响及其作用机制。分别配制浓度为10-6mol/L(W1)、10-4mol/L(W2)的ATRA溶液及浓度为100μg/ml(Z1)、200μg/ml(Z2)的SBA-Na溶液,分别使用W1、W2、Z1、Z2、W1+Z1、W2+Z2处理上述2种细胞系,并设立不加药的空白对照组。镜下观察不同处理组细胞形态及生长情况;应用CCK-8法检测细胞增殖能力,绘制细胞生长抑制曲线;分别采用PI单染和PI/Annexin V双染流式细胞术检测各加药组K562细胞和Kasumi-1细胞的细胞周期及凋亡的变化;采用实时荧光定量PCR(RQ-PCR)法检测K562细胞系各组CyclinA基因表达量的变化。结果表明,ATRA及SBA-Na对2种细胞均有抑制增殖作用,两者联合用药的作用更明显;各给药组与对照组相比,细胞周期分布发生明显变化,处理组细胞凋亡明显增加,尤以ATRA联合SBA-Na给药组凋亡最为明显。低浓度SBA-Na处理组Cyclin A表达上调,其他处理组Cyclin A表达均下调,且存在量效关系。结论:ATRA及SBA-Na均可抑制K562细胞及Kasumi-1胞系增殖,促进其凋亡,且二者联合效果更明显,对于K562细胞系,两者可能是通过下调Cyclin A基因表达而发挥作用。

Inhibitory Effect of All-Trans Retinoic Acid Combined with SBANa on K562 and Kasumi-1 Cell Lines in Vitro

This study was aimed to investigate the effect of all-trans retinoic acid（ATRA） combined with SBA-Na on the biologic activities of human leukemia K562 and Kasumi-1 cell lines and their mechanism.The ATRA solution of 10^-6mol/L（W1）,10^-4mol/L（W2） and the SBA-Na solution of 100 μg/ ml（Z1） and 200 μg/ml（Z2） were prepared respectively.The K562 and Kasumi-1 cells were treated with W1,W2,Z1,Z2,W1＋Z1 and W2＋Z2 respectively,at same time,the blank control was set up.The cell morphology and growth in different treated groups were observed under light microscope.The CCK-8 method was used to detect the proliferation ability of cells,the cell growth curves were drawn,the inhibitory rate of cells was calculated.The flow cytomertry with PI single staining and PI/Annexin V double stainings was used to detect the change of cell cycle and apoptosis of 2 cell lines treated with different drugs.The RQ-PCR was used to detect the change of Cyclin A mRNA expression in K562 cells.The results showed both ATRA and SBA-Na displayed inhibitory effect on cell proliferation,and the combination of these two drugs had stronger effect.As compared with the control group,the cell cycle distribution were changed obviously,and the apoptosis increased more significantly in treated groups,especially in group of ATRA combined with SBA-Na.The Cyclin A mRNA expression was up-regulated in Z1 group,while Cyclin A mRNA expression was down-regulated in other groups.It is concluded that both ATRA and SBA-Na can inhibit the proliferation of K562 and Kasumi-1 cell lines and promote their apoptosis.This effect may be stronger when both drugs combined.For K562 cells,the inhibitory effect may be accomplished through down-regulation of Cyclin A mRNA.