雷公藤内酯醇联合高三尖杉酯碱对KG-1α细胞增殖和凋亡的影响

Effects of Triptolide Combined with Homoharringtonine on Proliferation and Apoptosis of KG-1α Cells

目的:探讨低浓度雷公藤内酯醇(triptolide,TPL)联合高三尖杉酯碱(homoharringtonine,HHT)对KG-1α细胞增殖与凋亡的影响及其可能机制。方法:应用CCK-8法检测不同浓度TPL和HHT单用及联用对KG-1α细胞增殖的影响,并计算其药物联合作用指数(combined index,CI);甲基纤维素集落形成实验检测细胞集落形成能力;流式细胞术检测细胞表面分子、细胞凋亡率及细胞周期变化;Western blot检测Akt信号通路相关蛋白在TPL及HHT作用前后的表达情况。结果:KG-1α细胞高表达CD34CD123而CD38缺失;TPL及HHT单用对KG-1α细胞增殖抑制并具剂量依赖性(r=0.952,r=0.992);低浓度联合用药与单药对比,前者细胞增殖、集落形成率更低,而细胞凋亡率增高;CI值亦提示低浓度TPL联合HHT呈高度协同作用。低浓度TPL联合HHT作用KG-1α细胞后,P-Akt^(Ser473)、P-Akt^(Thr308)、BCL-2、PARP及Survivin蛋白表达降低而PARP裂解增加。结论:低浓度TPL联合HHT可通过PI3K/Akt信号通路及下游蛋白协同抑制KG-1α细胞增殖,并诱导其凋亡。

Objective： To investigate the effect and possible mechanism of low concentration of triptolide （TPL） combined with homoharringtonine （HHT） on the proliferation and apoptosis of KG-1 a cells. Methods： CCK-8 method was used to detect the antiproliferating effects of different concentrations of TPL and HHT single-use and combined use on KG-la cells, and the combined index （CI） was calculated. The colony formation ability was also determined by methylceUulose colony formation assay, cell surface molecules, apoptosis rate and cell cycle changes were detected by flow cytometry. Westerrn blot was used to detect the expression of Akt signaling pathway related proteins before and after low dose TPL combined with HHT using. Results： High expression of CD34 and CD123 were on KG-la cells, which being lack expression of CD38. TPL and HI-IT dose-dependently inhibited the proliferation of KG-1α cells. Compared with low dosage TPL and HHT single-use groups, the cell proliferation and colony formation efficiency were lower, and the cell apoptosis rate was higher in the combined group. CI values also indicated that low concentration TPL combined with HI-IT possessed highly synergistic effect. After the combination of the 2 drugs, the expressions of P-Akt^ser473 , P-Akt^thr308 , BCL-2, PARP and survivin protein were down-regulated and the cleavage of PARP protein was increased. Conclusion： Low concentration of TPL combined with HHT can synergistically inhibit KG-1α cell proliferation and induce its apoptosis through the PI3K/Akt signaling pathway and downstream protein.