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目的探讨青蒿琥酯对体外培养的人多发性骨髓瘤细胞株RPMI8226的增殖、凋亡及对凋亡相关基因survivin及caspase-3、caspase-7表达的影响。方法采用MTT法检测青蒿琥酯对RPMI8226细胞增殖的抑制作用,光镜下观察细胞形态学变化,流式细胞仪分析细胞凋亡率,荧光定量PCR(FQ-PCR)检测survivin和caspase-3、caspase-7mRNA表达水平变化,Western blotting检验Survivin蛋白表达变化,应用Caspase-3/7试剂盒检测Caspase-3、Caspase-7蛋白活性。结果青蒿琥酯质量浓度从2.5μg/mL增加至50μg/mL时,青蒿琥酯体外对RPMI8226细胞的抑制率由33.55%增加到74.71%。细胞凋亡率呈剂量依赖性增加,50μg/mL青蒿琥酯诱导RPMI8226细胞最大凋亡率达(68.1±5.9)%。不同质量浓度青蒿琥酯干预48h后Survivin mRNA及蛋白表达水平明显降低,Caspase-3、Caspase-7mRNA及蛋白活性明显升高(P<0.01)。结论青蒿琥酯对RPMI8226细胞有生长抑制和诱导凋亡作用,其机制可能与增强Caspase-3、Caspase-7活性,抑制抗凋亡分子survivin表达有关。
Objective To investigate the effects of artesunate on proliferation, apoptosis and the expression of survivin, caspase-3 and caspase-7 in human multiple myeloma cell line RPMI8226 cultured in vitro. Methods MTT assay was used to detect the inhibitory effect of artesunate on the proliferation of RPMI8226 cells. The morphological changes of the cells were observed under light microscope. The apoptosis rate was analyzed by flow cytometry. The expressions of survivin and caspase-3 were detected by fluorescence quantitative PCR (FQ-PCR) , Caspase-7mRNA expression levels were detected by Western blotting Survivin protein expression changes, the Caspase-3/7 kit Caspase-3, Caspase-7 protein activity. Results Artesunate increased the inhibitory rate of artesunate to RPMI8226 cells from 33.55% to 74.71% when the mass concentration of artesunate was increased from 2.5μg / mL to 50μg / mL. Apoptosis rate in a dose-dependent manner, 50μg / mL artesunate induced apoptosis rate of RPMI8226 cells (68.1 ± 5.9)%. The expression of Survivin mRNA and protein were significantly decreased and the mRNA and protein expressions of Caspase-3, Caspase-7 were significantly increased after artesunate treatment at different concentrations for 48h (P <0.01). Conclusion Artesunate can inhibit the growth and induce apoptosis of RPMI8226 cells. The mechanism may be related to the enhancement of the activity of Caspase-3 and Caspase-7 and the inhibition of the expression of survivin.