青蒿琥酯对硼替佐米耐药的骨髓瘤细胞株增殖与凋亡的影响  

Effects of artesunate on the proliferation and apoptosis of bortezomib-resistant multiple myeloma cells

作　　者：欧瑞明[1] 周长华[1] 谭友平[1] 刘爽[1] 钟启[1] 张青[1] OU Rui- ming;ZHOU Chang- hua;TAN You - ping;LIU Shuang;ZHONG Qi;ZHANG Qing(Department of Hematology, Guangdong Second Provincial General Hospital, Guangzhou 510317, China)

机构地区：[1]广东省第二人民医院血液科,广州510317

出　　处：《中国临床药理学杂志》2018年第10期1183-1186,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81400168);广东省中医药局科研基金资助项目(20131104);广东省医学科研基金资助项目(A2013128);广东省第二人民医院人才引进基金资助项目(YY2014-002)

摘　　要：目的研究青蒿琥酯对硼替佐米耐药的骨髓瘤细胞株的细胞增殖、细胞周期、细胞凋亡和细胞凋亡相关蛋白的影响。方法用硼替佐米浓度梯度递增法处理人骨髓瘤细胞株NCI-H929,建立硼替佐米耐药的NCI-H929细胞株NCI-H929BR。取对数生长期NCI-H929BR细胞,分为对照组(加入RPMI^(-1)640培养液)和低、高2个浓度实验组(25,50μg·m L^(-1)青蒿琥酯),分别处理48 h。用四甲基偶氮唑蓝(MTT)法检测NCI-H929BR的增殖情况;以流式细胞术检测细胞周期和细胞凋亡变化;以免疫印迹法检测B细胞淋巴瘤/白血病-2(Bcl-2)蛋白家族(Bcl-2、Bcl-Xl、Bax)的表达。结果用浓度梯度递增法成功建立了硼替佐米耐药的NCI-H929细胞株NCI-H929BR,其耐药指数为20.12倍。药物处理48 h后,25,50μg·m L^(-1)实验组的细胞增殖率分别为62.46%,33.48%,2个浓度实验组与对照组比较,差异均有统计学意义(均P<0.05);高浓度实验组与低浓度实验组比较,差异有统计学意义(P<0.05),说明抑制效应呈浓度依赖性。高浓度实验组处理细胞48 h,NCI-H929BR细胞的G1期细胞增加,G1期细胞百分比为(51.63±4.02)%;对照组为(34.72±2.43)%,实验组与对照组比较,差异有统计学意义(P<0.05)。高浓度实验组处理细胞24h,早期凋亡细胞百分比为(13.10±1.53)%,对照组为(2.32±1.02)%;晚期凋亡细胞百分比为(10.26±2.42)%,对照组为(1.93±0.69)%,实验组与对照组比较,差异均有统计学意义(均P<0.05)。低、高2个浓度实验组能降低Bcl-Xl、Bcl-2蛋白的相对表达量而升高Bax蛋白的表达,低、高2个浓度实验组与对照组比较,差异均有统计学意义(均P<0.05);高浓度实验组与低浓度实验组比较,差异有统计学意义(P<0.05),说明药物对蛋白表达的作用呈浓度依赖性。结论青蒿琥酯可抑制NCI-H929BR的增殖,促进其凋亡,产生G_0/G_1期阻滞,并下调Bcl-2、Bcl-Xl而上调Bax蛋白的表达。Objective To explore the effects of artesunate on the proliferation,cell cycle,apoptosis and apoptosis-related protein of bortezomib-resistant multiple myeloma(MM） cells. Methods Human MM cell line NCI-H929 was treated with bortezomib in a dose-dependent manner to establish a bortezomib-resistant cell line NCI-H929 BR. The logarithmic growth phase NCI-H929 BR cells were divided into control group(blank culture medium） and two concentration experimental groups(artesunate,25,50 μg·m L-1） and treated for 48 h. The inhibitory role of artesunate on NCI-H929 BR proliferation was measured by MTTassay. Cell apoptosis and cell cycle were determined by flow cytometry. The Bcl-2 family proteins(Bcl-2,Bcl-Xl,Bax） were detected by Western blot. Results Bortezomib resistance index of NCI-H929 BR is 20. 12 times than NCI-H929. After administration artesunate for 48 h,the proliferation rate of NCI-H929 BR in the low and high concentration experimental groups were 62. 46%,33. 48%; there was significant difference compared with control group(all P〈0. 05）. The artesunate dramatically decreases NCI-H929 BR proliferation in a dose-dependent manner(P〈0. 05）. After treatment for 48 h with artesunate,the proportion of G1 phase cell cycle in high concentration experimental group was increased compared with control group[(51. 63 ± 4. 02） % vs(34. 72 ± 2. 43） %,P〈0. 05]. The proportion of apoptosis cell was increased,early apoptosis in high concentration experimental group and control group were(13. 10 ± 1. 53） %,(2. 32 ± 1. 02） %; and late apoptosis in the two groups were(10. 26 ± 2. 42） %,(1. 93 ± 0. 69） %; comparison between two groups,the difference had significantly(all P〈0. 05）. In addition,artesunate significant decreased of Bcl-2 and Bcl-Xl protein expression and increased of Bax protein expression in a dose-dependent manner. Conclusion Artesunate could inhibit NCI-H929 BR proliferation,increase apoptosis,cause G0/G1-arrest and down-regulate Bcl-2 and Bcl-Xl expression,while up-regulate Bax

关 键 词：青蒿琥酯 硼替佐米 骨髓瘤 耐药 凋亡 

分 类 号：R28[医药卫生—中药学]