三氧化二砷与硼替佐米联合应用对多发性骨髓瘤KM3细胞凋亡的作用及其机制研究(英文)  

Effects of Arsenic Trioxide Combined with Bortezomib on Apoptosis of Multiple Myeloma Cell Line KM3 and Its Mechanisms

作　　者：葛群芳[2] 欧阳桂芳[2] 陈莹[1] 张怡[1] 牧启田[1] 陆滢[2] 

机构地区：[1]宁波市第一医院血液科 [2]宁波市第一医院,中心实验室,浙江宁波315010

出　　处：《中国实验血液学杂志》2012年第1期112-115,共4页Journal of Experimental Hematology

摘　　要：本研究探讨硼替佐米(Bor)单用及与三氧化二砷(As2O3)联合应用对多发性骨髓瘤(MM)细胞株KM3细胞凋亡的影响及其相关机制。采用MTT法检测Bor单用及与As2O3联合应用对KM3细胞增殖的抑制作用,计算其IC50值。采用AnnexinⅤ-FITC凋亡试剂盒检测各药物组KM3细胞早期及晚期凋亡率,流式细胞术测定各药物组KM3细胞跨膜电位的变化。采用RT-PCR检测各药物组KM3细胞Caspase-3、Bim、Bcl-xL mRNA水平的变化。结果表明,Bor联合As2O3对KM3细胞生长抑制明显高于Bor单用,处理72 h作用最明显〔(27.64±0.81)%vs(21.67±2.20)%,P<0.05〕。联合用药组KM3细胞早期凋亡率及晚期凋亡率均较Bor单药组明显增多,前者在作用48 h时效果最明显〔(53.20±3.70)%vs(35.40±2.58)%,P<0.01〕,后者在作用72 h时效果最明显〔(63.96±2.97)%vs(54.08±3.76)%,P<0.01〕。联合用药组KM3细胞线粒体跨膜电位较Bor单药组明显下降,作用48 h时下降最明显。与Bor单药组比较,联合用药组KM3细胞Caspase-3 mRNA、Bim mRNA均升高,Bcl-xLmRNA下降。结论:在体外As2O3可增强Bor对KM3细胞增殖的抑制,增加KM3细胞的凋亡,并且可能通过抑制Bcl-xL mRNA表达,诱导Caspase-3和BIM mRNA表达的途径增强凋亡作用。This study was purposed to investigate the effect of bortezomib （Bor） and arsenic trioxide （ AS2 03 ） com- bination on multiple myeloma cell line KM3 and its mechanisms. KM3 cells were cultured with different concentration of Bor or As203 as well as both for a certain time. The cell proliferation was analysed by MTT assay and the concentration of 50% proliferation inhibition （ IC50 ） was calculated. Early apoptosis and late apoptosis of KM3 cells were detected by Annexin-V-FITC Kit, and the change of transmembrane potential was measured by flow cytometry, mRNA of Caspase-3, Bim and Bcl-xL were detected by RT-PCR. The results showed that the proliferation inhibitory rate of KM3 cells treated by Bor plus As203 was much higher than that of KM3 cells treated by Bor only for 72 h （27.64 ±0.81）% vs （21.67 ± 2.20）%, P 〈 0.05 ~. There were more KM3 cells treated by Bor plus AszO3 in early apoptosis at 48 h and late apoptosis at 72 h than that of KM3 cells treated only by Bor [ （53.20 ± 3.70） % vs （ 35.40 ±2.58） %, P 〈 0. 01 ; （63.96± 2.97） % vs （54.08 ± 3.76 ） %, P 〈 0.01 ]. Transmembrane potential （AOm） of KM3 cells treated by Bor plus As203 decreased more at 48 h, as compared with Bor alone. The expression levels of caspase-3 mRNA and Bim mRNA in KM3 cells treated with Bor plus As203 were higher than that in KM3 cells treated with Bor alone. But the ex- pression level of Bcl-xL mRNA was lower than that in KM3 cells treated with Bor alone. It is concluded that As2 O3 can enhance the apoptosis-inducing effect of Bor on multiple myeloma cell line KM3, which is associated with decreasing the expression of Bcl-xl mRNA and increasing the expression of Caspase-3 and Bim mRNA.

关 键 词：三氧化二砷 硼替佐米 多发性骨髓瘤 KM3细胞 细胞凋亡 

分 类 号：R979.13[医药卫生—药品]