酪氨酸激酶抑制剂联合高三尖杉酯碱对慢性粒细胞白血病细胞系K562的影响

摘要

Objective:To investigate the effects of imatinib and nilotinib combined with homoharringtonine( HHT) on chronic myeloid leukemia cell line K562. Methods:Total 10 groups for the experiment:Control group,IM group, NL group,HHT group,HHT first followed by IM group,IM first followed by HHT group,HHT first followed by NL group,NL first followed by HHT group,IM simultaneously with HHT group and NL simultaneously with HHT group. CCK-8 was used to detect the viability of K562 cells,flow cytometer( FCM) to detect cell apotosis rate,Annexin V and PI double staining and western blot to detect the expression of BCR-ABL oncoprotein. Results:The apoptosis rate of experimental group was higher than that of control group. Among them,apoptotic rate in IM+HHT group was the highest,HHT-IM group second while there was no difference between IM-HHT group and single drug groups. Further study showed that,the expression of BCR-ABL protein in IM+HHT group was significantly reduced,HHT-IM was the second place while there was no difference between IM-HHT group and single drug groups. The same trend was ob-served when NL was combined with HHT. Conclusion:Different combined regimen of TKI and HHT can promote cell apoptosis and downregulate the expression of BCR-ABL oncoprotein. TKI combined with HHT simultaneously had the best effect,HHT followed by TKI treatment second.%目的：：探索伊马替尼( IM)、尼罗替尼( NL)这两种药物分别与高三尖杉酯碱( HHT)联合应用对慢性粒细胞白血病( CML)细胞系K562增殖、凋亡以及BCR-ABL癌蛋白表达的影响。方法：实验分为十个组,分别为：不加药对照组、IM单药组、NL单药组、HHT单药组、先HHT后IM组( HHT-IM)、先IM后HHT组( IM-HHT)、先HHT后NL组( HHT-NL)、先NL后HHT组( NL-HHT)、IM与HHT共同作用组( IM+HHT)、NL与HHT共同作用组( NL+HHT)。应用CCK-8法检测各组K562细胞的增殖抑制率,Annexin V/PI双染流式细胞术检测各组细胞凋亡率,Western blot检测各组BCR-ABL癌蛋白表达。结果：各实验组凋亡率均较对照组升高。其中,IM+HHT组细胞凋亡率明显高于其他各组,HHT-IM组次之,而IM-HHT组凋亡率与单药组差别不大。进一步研究发现,IM+HHT组BCR-ABL蛋白的表达明显减少, HHT-IM 次之,而 HHT单药组、IM-HHT组BCR-ABL蛋白减少的并不明显。 NL+HHT组趋势与之一致。结论：酪氨酸激酶抑制剂( TKI)与HHT多种联合用药方案可以促进CML细胞凋亡并下调BCR-ABL癌蛋白表达。其中TKI与HHT同时用药效果最佳,先HHT作用后TKI序贯治疗效果次之。