Department of Hematology, The Second Hospital of Hebei Medical University, 215, Heping Xi Road, 050000, Shijiazhuang City, People's Republic of China.
Department of Hematology, Affiliated Haikou Hospital, Xiangya School of Medicine, Central South University, Haikou Municipal Hospital, 570208, Haikou, People's Republic of China.
Int J Hematol. 2009 Nov;90(4):513-521. doi: 10.1007/s12185-009-0409-z. Epub 2009 Aug 29.
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 (NFkappaB p65) protein in nucleus and the inhibitor of NFkappaB (IkappaBalpha) in the cytoplasm were measured by western blot analysis and ELISA to evaluate NFkappaB 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 G0/G1 phase, while decreased the proportion of cells in G2/M or S phase. Additionally, artesunate treatment decreased the level of NFkappaB p65 protein in the nucleus, while increased the level of IkappaBalpha protein in the cytoplasm. The present result is the first report to show that artesunate may be useful in the treatment of MM.
多发性骨髓瘤（MM）的初始治疗在过去十年中发生了重大转变，新型药物如沙利度胺、来那度胺和硼替佐米的引入改善了治疗效果。高剂量化疗和自体干细胞移植仍然是适合该治疗方案的 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（常用于治疗重症疟疾）对抑制小鼠骨髓瘤细胞系 SP2/0 增殖和诱导其凋亡的作用。采用噻唑蓝（MTT）法检测青蒿琥酯（ART）处理对 SP2/0 细胞增殖的抑制作用，通过流式细胞术分析 ART 诱导的细胞凋亡率和细胞周期变化。通过光镜和透射电镜观察 ART 诱导的 SP2/0 细胞凋亡形态变化。此外，通过琼脂糖凝胶电泳检测从 ART 处理的 SP2/0 细胞中提取的 DNA，观察到 DNA 梯带，这是凋亡的标志。通过 Western blot 分析和 ELISA 间接评估核因子 kappa B p65（NFkappaB p65）蛋白在核内和抑制剂 of NFkappaB（IkappaBalpha）在细胞质中的水平，以评估 NFkappaB p65 转录活性。结果表明，青蒿琥酯以剂量和时间依赖的方式抑制 SP2/0 细胞的增殖并诱导其凋亡。青蒿琥酯还增加了 SP2/0 细胞处于 G0/G1 期的比例，同时降低了处于 G2/M 期或 S 期的细胞比例。此外，ART 处理降低了核内 NFkappaB p65 蛋白的水平，同时增加了细胞质中 IkappaBalpha 蛋白的水平。这是首次报道青蒿琥酯可能对 MM 治疗有效。
