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目的研究葫芦素E对人多发性骨髓瘤细胞增殖和凋亡的影响,并观察葫芦素E与阿霉素的协同效应。方法采用CCK-8测定葫芦素E对4种多发性骨髓瘤细胞MM1.S、MM1.R、U266和RPMI8226的半数生长抑制率(GI50),并通过CalcuSyn协同效应分析软件,计算药物合用指数(CI),评价葫芦素E与阿霉素协同抗多发性骨髓瘤作用;流式细胞术检测葫芦素E对MM1.S、MM1.R的细胞周期、线粒体膜电位和细胞凋亡的影响;Western blot检测凋亡相关蛋白Caspase-3和PARP的变化,以及葫芦素E对IL-10刺激的RPMI8226细胞STAT3蛋白磷酸化的抑制作用。结果葫芦素E对4种多发性骨髓瘤细胞的GI50均在(12.3~161.7)nM范围内;葫芦素E导致多发性骨髓瘤细胞的SubG1期细胞比例、凋亡细胞比例和线粒体膜电位下降细胞比例均明显增加,并出现凋亡相关的活化蛋白如Cleaved Caspase-3和Cleaved-PARP。葫芦素E能有效抑制IL-10诱导的RPMI8226细胞STAT3磷酸化,并能与阿霉素协同抗多发性骨髓瘤。结论葫芦素E通过抑制STAT3蛋白磷酸化,诱导多发性骨髓瘤细胞凋亡,并能协同阿霉素抗多发性骨髓瘤。
Objective To study the effect of cucurbitacin E on the proliferation and apoptosis of human multiple myeloma cells and to observe the synergistic effect of cucurbitacin E and doxorubicin. Methods The half inhibitory rate (GI50) of cucurbitacin E on four multiple myeloma cells, MM1.S, MM1.R, U266 and RPMI8226, was determined by CCK-8. The synergistic effect analysis software CalcuSyn was used to calculate the drug combination index CI) to evaluate the synergistic effect of cucurbitacin E and doxorubicin on multiple myeloma. The effects of cucurbitacin E on the cell cycle, mitochondrial membrane potential and apoptosis of MM1.S and MM1.R were examined by flow cytometry. blot was used to detect the changes of apoptosis associated protein Caspase-3 and PARP, and the inhibition of cucurbitacin E on phosphorylation of STAT3 protein in IL-10-stimulated RPMI8226 cells. Results The cucurbitacin E (GI) of cucurbitacin E was (12.3 ~ 161.7) nM in all the four myeloma cells. The cucurbitacin E led to the decrease of the percentage of SubG1 phase cells, apoptotic cells and mitochondrial membrane potential in multiple myeloma cells The proportion increased significantly, and the apoptosis-related activation proteins such as Cleaved Caspase-3 and Cleaved-PARP. Cucurbitacin E can effectively inhibit STAT-3 phosphorylation induced by IL-10 in RPMI8226 cells and synergize with doxorubicin against multiple myeloma. Conclusion Cucurbitacin E can induce multiple myeloma cell apoptosis by inhibiting the phosphorylation of STAT3 protein and can cooperate with doxorubicin in the treatment of multiple myeloma.