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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。
The aim of this study was to investigate the effects of all-trans retinoic acid (ATRA) combined with sodium cholate (SBA-Na) on the biological activities of human leukemia K562 cell line and Kasumi-1 cell line and its mechanism of action . ATRA solution of 10-6mol / L (W1), 10-4mol / L (W2) and SBA-Na solution of 100μg / ml (Z1) and 200μg / ml W2, Z1, Z2, W1 + Z1, W2 + Z2. The two control groups were established. The morphology and growth of cells in different treatment groups were observed under light microscopy. Cell proliferation was measured by CCK-8 assay and cell growth inhibition curve was drawn. PI staining and PI / Annexin V staining were used to detect the expression of K562 Cell cycle and apoptosis of Kasumi-1 cells. The expression of CyclinA gene in K562 cell line was detected by real-time fluorescence quantitative PCR (RQ-PCR). The results showed that both ATRA and SBA-Na inhibited the proliferation of both cell lines, and the combined effect of the two drugs was more obvious. Compared with the control group, the distribution of cell cycle significantly changed and the apoptosis of the treated group was obvious Increase, especially ATRA combined SBA-Na administration group the most obvious apoptosis. The expression of Cyclin A in low concentration SBA-Na group was up-regulated, while the expression of Cyclin A in other groups was down-regulated, and there was a dose-response relationship. CONCLUSION: Both ATRA and SBA-Na can inhibit the proliferation and promote the apoptosis of K562 cells and Kasumi-1 cell lines, and their combined effect is more obvious. Both of them may play a role in down-regulating Cyclin A gene expression in K562 cell line effect.