2-甲氧基雌二醇联合硼替佐米对多发性骨髓瘤细胞凋亡的影响

目的 探讨2-甲氧基雌二醇(2-ME2)联合硼替佐米的抗多发性骨髓瘤(MM)效应及与聚集小体(aggresome)形成之间的相关性.方法 以MM细胞系RPMI 8226、NCI-H929、U266、SKO-007细胞作为研究对象,采用硼替佐米单药或联合2-ME2处理,通过等效应线图法分析两者是否具有抗MM协同效应;通过免疫荧光技术在同一组细胞群中研究聚集小体阳性和阴性细胞的凋亡情况.结果 ①硼替佐米呈浓度和时间依赖性地显著提高聚集小体阳性细胞比例,且聚集小体几乎全部出现于非凋亡细胞中.②等效应线图法分析显示一定浓度范围的硼替佐米与2-ME2联合具有显著的协同作用.③硼替佐米与2-ME2联合应用后聚集小体阴性的凋亡细胞显著增多而聚集小体阳性的非凋亡细胞明显减少.在RPMI 8226及U266细胞中,硼替佐米单药与联合用药组相比,聚集小体阴性的凋亡细胞分别由(14.5±2.6)%及(20.1±2.9)%增加至(80.7±6.9)%及(71.6±6.2)%;聚集小体阳性的非凋亡细胞分别由(75.3±5.7)%及(69.1±8.6)%减少到(13.8±3.8)%及(19.5±4.2)%.结论 硼替佐米诱导生成的聚集小体对MM细胞具有保护作用;2-ME2通过抑制聚集小体途径可以显著增强硼替佐米的抗MM效应.

2-methoxyestradiol disrupts aggresomes induction by bortezomib and potentiates apoptosis in multiple myeloma cells

Objective To investigate the synergistic anti-multiple myeloma(MM) effect of 2-methoxyestradiol(2-ME2) and bortezomib,and explore the relationship between this effect and blockade of aggresomes formation by 2-ME2.Methods Four MM cell lines RPMI-8226,NCI-H929,U266 and SKO-007 were used for study.Immunoflourescent anti-ubiquitin and Hoechst 33342 staining were used to examine aggresome-positive cells and apoptotic cells,respectively.Isobolographic analysis was used for determination of synergy.Results ①Quantitative assay showed that in the absence of bortezomib,only 6.6%-8.9% of MM cells were aggresome-pesitive,but the percentage was increased to 71.9%-83.4% after treatment with bortezomib at IC_(20) concentration for 24 h.Aggresome-positive cells with immunoreactivity to anti-ubiquitin were detected in almost all non-apoptotic ceils,but not in apoptotic cells. ②Treatment in a definite range of concentrations bortezomib plus 2-ME2 led to MM cell apeptosis compared with each agent alone and the signif-icantly synergistic effect confirmed by isobolographic analysis. ③Combination of bortezomib and 2-ME2 increased the apoptotic cells aggresome-negative cells(ANK) and decreased the non-apoptotic cells in aggresome positive cells (APC).In RPM18226 and U266 cells,the apoptotic cells in ANC increased from (14.5±2.0)% and (20.1±2.9)% to (80.7±6.9)% and (71.6±6.2)%,and the non-apoptotic cells in APC decreased from (75.3±5.7)% and (69.1±8.6)% to (13.8±3.8)% and (19.5±4.2)%,respectively,in combined group and bortezomib alone group.Conclusion Bortezomib-induced aggresomes have a protective function for MM cells and combination of bortezomib with 2-ME2 induced a synergistic cytotoxicity to the cells.