ROS在小白菊内酯诱导MM细胞凋亡中的作用

目的研究倍半萜内酯小白菊内酯(parthenolide,PL)对多发性骨髓瘤原代细胞和细胞株诱导的凋亡作用及其机制。方法以多发性骨髓瘤原代细胞及细胞株为研究对象,利用台盼蓝染色检测细胞存活率,用DAPI染色法检测细胞凋亡率,流式细胞仪检测反应氧(ROS)和线粒体膜电位水平变化。结果2.5μmol.L-1PL引起多发性骨髓瘤(multi-ple myeloma,MM)细胞明显凋亡及线粒体膜电位下降,而正常淋巴细胞在同等条件下不受影响。用2.5μmol.L-1PL作用10 h后,KMM-1和MM1S细胞ROS水平明显增加。用自由基清除剂L-N-乙酰半胱氨酸(L-NAC)预处理可抑制ROS的产生,对两种细胞存活率、凋亡率无影响。用还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶抑制剂二亚苯基烟碱氯化物(diphenylene iodonium chloride,DPI)预处理KMM-1和MM1S,可明显抑制PL介导的ROS的产生,并对两种细胞凋亡率无影响。结论PL诱导多发性骨髓瘤细胞凋亡,而正常淋巴细胞在同等条件下不受影响。PL的凋亡活性是通过引起MM细胞氧化应激而介导的,并进一步推测NADPH氧化活性与PL介导的MM细胞ROS的产生相关。

Effect of ROS on parthenolide-induced apoptosis in multiple myeloma cell

Aim To explore the effect of sesquiterpene lactone parthenolide(PL) on primary multiple myeloma(MM) cell and cell lines and its molecular mechanisms.Methods Fresh bone marrow cells from MM patients and MM cell lines KMM-1 and MM1S were used for in vitro culture study.The cell survival rate was examined by Trypan Blue exclusion.The change of nucleolus stained with DAPI was observed under fluorescent microscope.Reactive oxygen species(ROS) levels and mitochondrial membrane potential(MMP) were detected by flow cytometry.Results 2.5 μmol·L-1 PL induced MM cell apoptosis and MMP was profoundly decreased in KMM-1 and MM1S cells.In contrast,normal lymphocytes cells were unaffected by the same conditions.KMM-1 and MM1S cells showed large increases of ROS levels 10 h after 2.5 μmol·L-1 PL treatment.The free radical scavenger L-N-acetylcystein(L-NAC) pretreatment completely inhibited ROS generation,cell death and nuclear fragmentation by PL.Diphenylene iodonium chloride(DPI) pretreatment in a dose of 2.5 μmol·L-1 strongly inhibited PL-mediated ROS generation,nuclear fragmentation in KMM-1 and MM1S cells.Conclusions PL strongly induces apoptosis in MM cell,while normal lymphocytes cells are completely unaffected by the same conditions.Since its proapoptotic activity appears to be mediated through induction of oxidative stress in MM cells,its possible mechanism is that NADPH oxidase activation is involved in PL-mediated ROS generation.