葫芦素E抑制多发性骨髓瘤细胞增殖及与阿霉素的协同效应

The Antiproliferative Effect of Cucurbitacin E on Human Multiple Myeloma Cells and Its Synergistic Effect in Combination with Adriamycin

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 detect the effect of cucurbitacin E（CuE）on the proliferation and apoptosis of human mulitiple myeloma（MM）cells,and to investigate the synergistic effect of CuE combined with adriamycin on MM cells.Methods The CCK-8assay was used to measure the growth inhibition of 50%（GI50）of CuE on the four MM cell lines including MM1.S,MM1.R,U266 and RPMI8226.The synergistic effect of CuE combined with Adriamycin on MM cells was assessed by calculating combination index（CI）with the help of CalcuSyn software（Biosoft,Cambridge,UK）.The flow cytometer was used to measure the effect of CuE on cell cycles,mitochondrial membrane potential and cell apoptosis,and the Western-blot was adopted to detect the changes of Caspase-3and PARP and the inhibition effect of CuE on STAT3 phosphorylation induced by IL-10.Results The GI50 of CuE on the four MM cell lines ranged from 12.3nM to 161.7nM.CuE significantly increased the number of the cells in the subG1 phase and the percentage of apoptotic cells,but reduced the mitochondrial membrane potential of MM cells.Meanwhile,Cleaved Caspase-3and cleaved PARP were detected in MM cells.CuE had a great inhibition effect on the STAT3 phosphorylation in MM cells induced by IL-10 and a synergistic effect on human multiple myeloma in combination with Adriamycin.Conclusion CuE can induce the apoptosis of MM cells by inhibiting the phosphorylation of STAT3,and have a synergistic effect of anti-multiple myeloma by working with Adriamycin.