去甲斑蝥素通过NF—κB／IκBα途径增强阿霉素的抗骨髓瘤效应  

Norcantharidin potentialize the chemosensitivity of adriamycin through the NF-κB/IκBα signaling pathway

作　　者：宋晓宁[1] 杜恒飞[1] 于路佳 蒙艳凤[1] 吕鸿雁[1] 孙丽霞 孟建波[1] 张金巧[1] 

机构地区：[1]河北医科大学第三医院血液科,石家庄050051 [2]淄博市第一医院血液科

出　　处：《中华血液学杂志》2011年第12期809-813,共5页Chinese Journal of Hematology

基　　金：河北省中医药管理局科研课题（2007136）

摘　　要：目的研究去甲斑蝥素（NCTD）联合阿霉素（ADR）对多发性骨髓瘤细胞增殖、凋亡的影响及其机制。方法以U266细胞为研究对象，采用NCTD（10μmol／L）和ADR（0．25μmol／L）单独或联合处理细胞，MTT法观察细胞增殖活力，流式细胞术检测细胞凋亡率，Western blot法检测核因子-κBP65（NF—κBP65）、磷酸化NF—κBP65（p-NF—κBP65）、NF—κB抑制因子IκBα、磷酸化IκBα（p-IκBα）、survivin、Bcl-2和Bax蛋白水平，免疫组化法测定血管内皮细胞生长因子（VEGF）水平。结果①NCTD能增强ADR的细胞毒和诱导凋亡作用，二者具有协同作用；②与ADR单药组比较，联合用药组胞核NF—κBP65和胞质P—IκBα的表达量分别由2．08±0．29和0．39±0．07降至0．48±0．08和0．02±0．01，胞质NF—κBP65和IκBα表达无变化；③联合用药组与ADR单药比较，survivin和Bcl-2的表达水平分别由0．31±0．05和0．23±0．05降至0．03±0．02和0．05±0．02，而Bax的表达由0．46±0．06升至0．62±0．08；④ADR组和联合用药组VEGF的阳性率分别为（44．6±4．4）％和（27．0±2．1）％，NCTD增强ADR对VEGF表达的抑制作用。结论NCTD通过抑制NF—κB／IκBα途径，调节下游信号分子survivin、Bel-2、Bax和VEGF的表达，增强ADR的抗骨髓瘤效应。Objective To explore the synergetic effect of norcantharidin （NCTD） and adriamycin （ADR） on the proliferation and apoptosis of multiple myeloma （MM） ceils. Methods Human MM cell line U266 cells were treated with NCTD alone （10 μmol/L） or in combination with ADR （0.25 μmol/L）. MTT and Annexin V/PI staining were used to determine cell viability and apoptosis. The protein expression of nuclear factor-κB P65 （ NF-κB P65 ） , phosphorylated NF-κB p65 （ p-NF-κB p65 ） , NF-κB P65 inhibitor IκBα, phosphorylated IKBct （p-IκBα）, survivin, Bcl-2 and Bax were determined by Western blot. Immunohistochemistry was used to determine the expression of vascular endothelial growth factor （VEGF）. Results （1） NCTD potentiated the cytotoxicity and pro-apoptotic effects induced by ADR. The combination of NCTD and ADR had synergistic anti-proliferation effect. （2）Combination of ADR and NCTD downregulated the expression of nuclear NF-κB P65 and cytoplasm p-IκBα induced by ADR. The expression of nuclear NF-KB P65 and cy- toplasm p-IκBα decreased from 2.08 ±.29 and 0.39 ± 0.07 to 0.48 ± 0.08 and 0.02 ± 0.01 respectively, while the expression of the cytoplasm NF-κB P65 and IκBα were unchanged in the ADR alone group and the combined group. （3）The expression of survivin and bcl-2 decreased from 0.31 ±0.05 and 0.23 ± 0.05 to 0.03 ± 0.02 and 0.05 ± 0.02, while the expression of Bax increased from 0.46 ± 0.06 to 0.62 ±0.08 respectively in ADR alone group and combined group. （4）The positive rate of VEGF in ADR group and combination group were （44.6 ± 4.4）% and （27.0 ± 2.1 ）% respectively, indicating that NCTD could potentiate the inhibition effect on VEGF induced by ADR. Conclusions The results suggest that NCTD can potentialize the chemosensitivity of multiple myeloma cells to ADR through regulating NF-κB/IκBα signaling pathway and NF- κB-regulated gene products including survivin, Bcl-2, Bax and VEGF.

关 键 词：去甲斑蝥素 阿霉素 多发性骨髓瘤 NF—κB 

分 类 号：R733.3[医药卫生—肿瘤]