摘要
本研究旨在探讨熊果酸对急性髓系白血病t(8;21)阳性细胞株kasumi-1细胞的抑制增殖、诱导凋亡机制。采用CCK-8法检测细胞增殖抑制,瑞氏染色法观察细胞凋亡形态,流式细胞术检测细胞凋亡率,实时荧光定量PCR法及Western blot法检测kasumi-1细胞内相关基因mRNA及蛋白的表达。结果表明,经熊果酸处理后的kasumi-1细胞的生长增殖受到抑制,出现凋亡形态学改变;在熊果酸2.5、5、10、20、50μmol/L浓度梯度以及在12,24,36,48,60,72 h时间梯度下,Annexin V阳性细胞比率随药物浓度的增加及作用时间的延长呈上升趋势;细胞内AML1-ETO、KIT、MYC、CCND1、BCL2、MDM2 mRNA的表达随作用时间的延长及用药浓度的增加而呈显著的下降趋势,而P53及BAX的mRNA表达逐渐增高;Western blot显示AML1-ETO、C-kit、C-MYC、BCL-2蛋白表达减低,而P53蛋白表达增高。结论:熊果酸呈浓度和时间依赖性抑制kasumi-1细胞的增殖并诱导其凋亡,从而发挥抗白血病的作用。这为t(8;21)白血病靶向治疗方案的选择提供理论依据。
This study was purposed to investigate the anti-tumor effect of ursolic Acid （UA） on t （8;21） leukemia cell line kasumi-1 and its possible mechanismisms. The kasumi-1 cells were treated with UA at different concentration for different duration of time. The growth inhibition of kasumi-1 treted with UA was detected by using CCK-8 test, and the morphological changes of kasumi-1 cells were observed by Wright＇s staining. Furthermore, the apoptosis rate of kasumi- 1 was examined by flow cytometry. Lastly, the expression of AML1-ETO, KIT, MYC, CCND1, BCL-2, P53, BAX, MDM2 and protein were detected by using real-time quantitative PCR and Western blot respectively. The results showed that the UA obviously inhibited the growth of kasumi-1 cells in dose- and time-dependent manners. The apototic morphological changes of cells were presented when kasumi-1 cells were treated with UA for 48 hours. The apoptotic rate of kasumi-1 cells increased in a dose- and time-dependent ways, and the mRNA levels of AML1-ETO, KIT, MYC, CCND1, BCL2, MDM2 decreased in kasumi-1 cells treated with UA, as well as the protein levels. Meanwhile, UA upregulated the mRNA and protein levels of P53 in the same manner. It is concluded that UA can exert its anti-tumor effect by inhibiting the proliferation and inducing the apoptosis of kasumi-1 cells in a dose-and time-dependent manners, that may provide the clues for a new targeting therapy to t（8 ;21 ） leukemia.
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2014年第3期687-691,共5页
Journal of Experimental Hematology
关键词
熊果酸
KASUMI-1
增殖抑制
诱导凋亡
ursolic acid
kasumi-1
inhibition of proliferation
induction of apoptosis
