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)

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。

Effects of arsenic trioxide combined with bortezomib on apoptosis of multiple myeloma cell line KM3 and its mechanisms

This study was purposed to investigate the effect of bortezomib (Bor) and arsenic trioxide (As(2)O(3)) combination on multiple myeloma cell line KM3 and its mechanisms. KM3 cells were cultured with different concentration of Bor or As(2)O(3) as well as both for a certain time. The cell proliferation was analysed by MTT assay and the concentration of 50% proliferation inhibition (IC(50)) 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 As(2)O(3) 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 As(2)O(3) 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 (Δψm) of KM3 cells treated by Bor plus As(2)O(3) 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 As(2)O(3) were higher than that in KM3 cells treated with Bor alone. But the expression level of Bcl-xL mRNA was lower than that in KM3 cells treated with Bor alone. It is concluded that As(2)O(3) 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.