Effect of artesunate on inhibiting proliferation and inducing apoptosis of SP2/0 myeloma cells through affecting NFκB p65
Effect of artesunate on inhibiting proliferation and inducing apoptosis of SP2/0 myeloma cells through affecting NFκB p65
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DOI：
10.1007/s12185-009-0409-z
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发表时间：
2009-11-01
影响因子：
2.1
通讯作者：
Pan, Ling
中科院分区：
文献类型：
--
作者：
Li, Shihui;Xue, Fang;Pan, Ling
在过去的十年中，随着新型药物如沙利度胺、来那度胺和硼替佐米的引入，多发性骨髓瘤（MM）的初始治疗经历了范式转变，从而改善了结局。高剂量治疗和自体干细胞移植仍然是适合该手术的MM患者的重要治疗选择。然而，这些治疗方案中的大多数对中国患者来说过于昂贵。因此，我们研究了青蒿琥酯，这是常用的治疗严重疟疾，对小鼠骨髓瘤细胞系SP2/0的增殖抑制和诱导凋亡的影响。采用MTT法检测青蒿琥酯（ART）对SP2/0细胞增殖的抑制作用，流式细胞仪检测ART诱导的细胞凋亡率和细胞周期变化。光镜和透射电镜观察ART诱导SP2/0细胞凋亡的形态学变化。此外，DNA梯状化，这是细胞凋亡的标志，用琼脂糖凝胶电泳法检测ART处理的SP 2/0细胞DNA的表达，核因子kappa B p65的表达水平在ART处理后的SP 2/0细胞中明显高于对照组核内NF-κ B p65蛋白及其抑制剂通过蛋白质印迹分析和ELISA测量细胞质中的（I kappa B alpha）以间接评估NF kappa B p65转录活性。结果表明，青蒿琥酯可抑制SP2/0细胞的增殖并诱导其凋亡，且呈剂量和时间依赖性。青蒿琥酯还可增加SP 2/0细胞的G（0）/G（1）期细胞比例，减少G2/M期或S期细胞比例。此外，青蒿琥酯处理降低核中NF κ B B p65蛋白的水平，而增加细胞质中I κ B α蛋白的水平。本研究结果是首次报道青蒿琥酯治疗多发性骨髓瘤。
The initial treatment of multiple myeloma (MM) experienced a paradigm shift, in the past decade, with the introduction of novel agents such as thalidomide, lenalidomide and bortezomib, leading to improved outcomes. High dose therapy and autologous stem cell transplantation remain an important therapeutic option for patients with MM eligible for the procedure. However, most of these treatment regimens are too expensive for Chinese patients. Therefore, we investigated the effects of artesunate, which is commonly used in the treatment of severe malaria, on inhibition of proliferation and induction of apoptosis of a mouse myeloma cell line SP2/0. The growth inhibition of SP2/0 cell proliferation induced by artesunate (ART) treatment was measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method and the rate of apoptosis and cell cycle changes induced by ART were analyzed by flow cytometry. ART-induced morphology changes of apoptosis in SP2/0 cells, as observed by light and transmission electron microscopy. Additionally, DNA laddering, which is a hallmark of apoptosis, was observed by agarose gel electrophoresis of DNA harvested from SP2/0 cells treated with ART. The levels of nuclear factor kappa B p65 (NF kappa B p65) protein in nucleus and the inhibitor of NF kappa B (I kappa B alpha) in the cytoplasm were measured by western blot analysis and ELISA to evaluate NF kappa B p65 transcription activity indirectly. The results show that artesunate inhibited the proliferation and induced apoptosis of SP2/0 cells in a dose- and time-dependent manner. Artesunate also increased the proportion of SP2/0 cells in G(0)/G(1) phase, while decreased the proportion of cells in G2/M or S phase. Additionally, artesunate treatment decreased the level of NF kappa B p65 protein in the nucleus, while increased the level of I kappa B alpha protein in the cytoplasm. The present result is the first report to show that artesunate may be useful in the treatment of MM.