摘要
目的探讨蛋白酶体抑制剂硼替佐米单用及与三氧化二砷或高三尖杉酯碱联合作用对髓系白血病细胞株HL-60的凋亡诱导作用。方法MTT法、Hoechst33342染色分别观察细胞增殖抑制及细胞凋亡情况，Western印迹检测bcl-2、Caspase-9、Caspase-3、PARP蛋白表达。结果硼替佐米、三氧化二砷、高三尖杉酯碱单独作用时均对HL-60细胞有明显的增殖抑制作用及凋亡诱导作用；三氧化二砷或高二三尖杉酯碱与硼替佐米联合对HL-60细胞的增殖抑制作用比3种药物单独作用增强（均P〈0.05）；形态学观察显示药物联合作用后凋亡诱导作用也比单药作用增强。Western印迹法检测显示15μmol／L二三氧化二砷单独作用细胞后，Caspase-9、Caspase-3、PARP均出现裂解片段，bcl-2蛋白表达水平下降；30nmol／L高三尖杉酯碱单药作用后PARP出现裂解片段、bcl-2蛋白表达水平下降，但是Caspase-9、Caspase-3的表达与对照组比较无改变；联合用药后相关蛋白的改变与细胞凋亡相平行。结论高三尖杉酯碱或三氧化二砷与硼替佐米联合后对细胞的凋亡诱导有相加作用。三氧化二砷与硼替佐米的凋亡诱导相加作用与二者能共同抑制Caspase信号途径及bcl-2蛋白的表达有关，高三尖杉酯碱和硼替佐米的相加作用与二者共同抑制bcl-2蛋白表达及促进PARP裂解活化有关。
Objective To explore the apoptosis effect induced by bortezomib combined with homoharringtonine or arsenious acid in HL-60 cell line and the mechanism. Methods Cell＇ s apoptosis was demonstrated by MTT assay and Hoechst33342 staining. Expression of bel-2, Caspase-9, Caspase-3 and PARP protein was detected by Western blot. Results HL-60 cells＂ apoptosis could be induced by bortezomib, homoharringtonine and arsenious acid respectively. Proliferation inhibition of HL-60 cells could be enhanced significantly when treated by bortezomib combined witl, homoharringtonine or arsenious acid compared with treated by any of the three drugs alone （P 〈 0.05）. At the same time morphology shows the apoptosis induced by drugs combined is more obviously than by one drug. Western blot showed bel-2 protein was down-regulated and Caspase-9, Caspase-3 and PARP proteins were all cleaved activation when cells were treated by 15 μmol/ L arsenious acid alone, but only cleaved activation of PARP and down-regulation of bcl-2 protein be detected when cells were treated with 30 nmol/L homoharringtonine alone, expression of Caspase-9 and Caspase-3 had no change compared with the control. The changes of associated proteins were paralleled with the cell＂ s apoptosis when treated with combined drugs. Conclusion HL-60 cells＂ apoptosis effect is inhaneed significantly when bortezomib combined with homoharringtonine or arsenious acid. Arsenious acid and bortezomib can inhibit caspase signaling pathway and down-regulate the expression of bcl-2 protein together, but homoharringtonine and bortezomib can only down-regulate the expression of bcl-2 protein and induce cleaved activation of PARP together.
出处
《白血病．淋巴瘤》
CAS
2013年第9期528-531,共4页
Journal of Leukemia & Lymphoma
