中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2011年
12期
809-813
,共5页
宋晓宁%杜恒飞%于路佳%蒙艳凤%吕鸿雁%孙丽霞%孟建波%张金巧
宋曉寧%杜恆飛%于路佳%矇豔鳳%呂鴻雁%孫麗霞%孟建波%張金巧
송효저%두항비%우로가%몽염봉%려홍안%손려하%맹건파%장금교
去甲斑蝥素%阿霉素%多发性骨髓瘤%NF-κB
去甲斑蝥素%阿黴素%多髮性骨髓瘤%NF-κB
거갑반모소%아매소%다발성골수류%NF-κB
Norcantharidin%Adriamycin%Multiple myeloma%NF-κB
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(10 μmol/L)和ADR(0.25 μmol/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-κB P65( NF-κB P65)、磷酸化NF-κB P65( p-NF-κB P65)、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-κB P65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-κB P65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、Bcl-2、Bax和VEGF的表达,增强ADR的抗骨髓瘤效应.
目的 研究去甲斑蝥素(NCTD)聯閤阿黴素(ADR)對多髮性骨髓瘤細胞增殖、凋亡的影響及其機製.方法 以U266細胞為研究對象,採用NCTD(10 μmol/L)和ADR(0.25 μmol/L)單獨或聯閤處理細胞,MTT法觀察細胞增殖活力,流式細胞術檢測細胞凋亡率,Western blot法檢測覈因子-κB P65( NF-κB P65)、燐痠化NF-κB P65( p-NF-κB P65)、NF-κB抑製因子IκBα、燐痠化IκBα(p-IκBα)、survivin、Bcl-2和Bax蛋白水平,免疫組化法測定血管內皮細胞生長因子(VEGF)水平.結果 ①NCTD能增彊ADR的細胞毒和誘導凋亡作用,二者具有協同作用；②與ADR單藥組比較,聯閤用藥組胞覈NF-κB P65和胞質p-IκBα的錶達量分彆由2.08±0.29和0.39±0.07降至0.48±0.08和0.02±0.01,胞質NF-κB P65和IκBα錶達無變化；③聯閤用藥組與ADR單藥比較,survivin和Bcl-2的錶達水平分彆由0.31±0.05和0.23±0.05降至0.03±0.02和0.05±0.02,而Bax的錶達由0.46±0.06升至0.62±0.08；④ADR組和聯閤用藥組VEGF的暘性率分彆為(44.6±4.4)％和(27.0±2.1)％,NCTD增彊ADR對VEGF錶達的抑製作用.結論 NCTD通過抑製NF-κB/IκBα途徑,調節下遊信號分子survivin、Bcl-2、Bax和VEGF的錶達,增彊ADR的抗骨髓瘤效應.
목적 연구거갑반모소(NCTD)연합아매소(ADR)대다발성골수류세포증식、조망적영향급기궤제.방법 이U266세포위연구대상,채용NCTD(10 μmol/L)화ADR(0.25 μmol/L)단독혹연합처리세포,MTT법관찰세포증식활력,류식세포술검측세포조망솔,Western blot법검측핵인자-κB P65( NF-κB P65)、린산화NF-κB P65( p-NF-κB P65)、NF-κB억제인자IκBα、린산화IκBα(p-IκBα)、survivin、Bcl-2화Bax단백수평,면역조화법측정혈관내피세포생장인자(VEGF)수평.결과 ①NCTD능증강ADR적세포독화유도조망작용,이자구유협동작용；②여ADR단약조비교,연합용약조포핵NF-κB P65화포질p-IκBα적표체량분별유2.08±0.29화0.39±0.07강지0.48±0.08화0.02±0.01,포질NF-κB P65화IκBα표체무변화；③연합용약조여ADR단약비교,survivin화Bcl-2적표체수평분별유0.31±0.05화0.23±0.05강지0.03±0.02화0.05±0.02,이Bax적표체유0.46±0.06승지0.62±0.08；④ADR조화연합용약조VEGF적양성솔분별위(44.6±4.4)％화(27.0±2.1)％,NCTD증강ADR대VEGF표체적억제작용.결론 NCTD통과억제NF-κB/IκBα도경,조절하유신호분자survivin、Bcl-2、Bax화VEGF적표체,증강ADR적항골수류효응.
Objective To explore the synergetic effect of norcantharidin (NCTD) and adriamycin (ADR) on the proliferation and apoptosis of multiple myeloma ( MM ) cells.Methods Human MM cell line U266 cells were treated with NCTD alone (10 μ mol/L) or in combination with ADR (0.25 μ mol/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 IκBα (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 ①NCTD potentiated the cytotoxicity and pro-apoptotic effects induced by ADR.The combination of NCTD and ADR had synergistic anti-proliferation effect.②Combination of ADR and NCTD downregulated the expression of nuclear NF-κB P65 and cytoplasm p-IκBα induced by ADR.The expression of nuclear NF-κB P65 and cytoplasm p-IκBα decreased from 2.08 ± 0.29 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.③The 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.④The 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 NFκB-regulated gene products including survivin,Bcl-2,Bax and VEGF.
