中华血液学杂志
中華血液學雜誌
중화혈액학잡지
Chinese Journal of Hematology
2012年
11期
911-916
,共6页
尚晋%陈志忠%吴文滨%魏天南%陈为民
尚晉%陳誌忠%吳文濱%魏天南%陳為民
상진%진지충%오문빈%위천남%진위민
硼替佐米%抗药性,多药%核转录因子%凋亡抑制蛋白
硼替佐米%抗藥性,多藥%覈轉錄因子%凋亡抑製蛋白
붕체좌미%항약성,다약%핵전록인자%조망억제단백
Bortezomib%Drug resistance,multiple%Nuclear transcription factor%Inhibitor of apoptosis protein
目的 观察硼替佐米对多药耐药白血病细胞增殖的影响及耐药逆转作用,初步探讨硼替佐米抑制多药耐药白血细胞增殖的分子机制.方法 以多药耐药白血病细胞系HL-60/DNR和HL-60/VCR细胞为模型,以HL-60细胞为对照,用四甲基偶氮唑盐反应比色法(MTT法)测定硼替佐米对HL-60、HL-60/DNR和HL-60/VCR细胞增殖的抑制作用,计算微毒剂量作为逆转耐药剂量.分4个实验组:HL-60/DNR+柔红霉素(DNR)、HL60/DNR+DNR+硼替佐米、HL-60/VCR+长春新碱(VCR)、HL-60/VCR +VCR+硼替佐米,计算硼替佐米逆转耐药倍数.硼替佐米按浓度递增(10、40、80nmol/L)作用于HL-60/DNR和HL-60/VCR细胞48 h,实时定量RT-PCR和Western blot方法检测XIAP、cIAP-1、cIAP-2 mRNA和蛋白表达水平及NF-κB活性.结果 硼替佐米呈浓度依赖性抑制HL-60、HL-60/DNR和HL-60/VCR细胞增殖,IC50值分别为(28.90 ±3.99)、(81.19 ±9.34)和(73.48±8.94) nmol/L.10 nmol/L硼替佐米预处理48 h后,DNR对HL-60/DNR细胞的IC50值由(12.90±1.75) μmol/L降低至(3.54±0.57) μmol/L(P ＜0.01),VCR对HL-60/VCR的IC50值由(33.25 ±7.28)μmol/L降低至(9.97±1.15) μmol/L(P ＜0.01),逆转耐药倍数分别为(3.32±0.53)和(2.64±0.28)倍.硼替佐米呈浓度依赖性下调XIAP、cIAP-1、cIAP-2 mRNA和蛋白的表达水平及抑制NF-κB活化.结论 硼替佐米可抑制多药耐药HL-60细胞增殖,并对多药耐药具有一定的逆转作用；其机制可能与下调凋亡抑制蛋白表达相关.
目的 觀察硼替佐米對多藥耐藥白血病細胞增殖的影響及耐藥逆轉作用,初步探討硼替佐米抑製多藥耐藥白血細胞增殖的分子機製.方法 以多藥耐藥白血病細胞繫HL-60/DNR和HL-60/VCR細胞為模型,以HL-60細胞為對照,用四甲基偶氮唑鹽反應比色法(MTT法)測定硼替佐米對HL-60、HL-60/DNR和HL-60/VCR細胞增殖的抑製作用,計算微毒劑量作為逆轉耐藥劑量.分4箇實驗組:HL-60/DNR+柔紅黴素(DNR)、HL60/DNR+DNR+硼替佐米、HL-60/VCR+長春新堿(VCR)、HL-60/VCR +VCR+硼替佐米,計算硼替佐米逆轉耐藥倍數.硼替佐米按濃度遞增(10、40、80nmol/L)作用于HL-60/DNR和HL-60/VCR細胞48 h,實時定量RT-PCR和Western blot方法檢測XIAP、cIAP-1、cIAP-2 mRNA和蛋白錶達水平及NF-κB活性.結果 硼替佐米呈濃度依賴性抑製HL-60、HL-60/DNR和HL-60/VCR細胞增殖,IC50值分彆為(28.90 ±3.99)、(81.19 ±9.34)和(73.48±8.94) nmol/L.10 nmol/L硼替佐米預處理48 h後,DNR對HL-60/DNR細胞的IC50值由(12.90±1.75) μmol/L降低至(3.54±0.57) μmol/L(P ＜0.01),VCR對HL-60/VCR的IC50值由(33.25 ±7.28)μmol/L降低至(9.97±1.15) μmol/L(P ＜0.01),逆轉耐藥倍數分彆為(3.32±0.53)和(2.64±0.28)倍.硼替佐米呈濃度依賴性下調XIAP、cIAP-1、cIAP-2 mRNA和蛋白的錶達水平及抑製NF-κB活化.結論 硼替佐米可抑製多藥耐藥HL-60細胞增殖,併對多藥耐藥具有一定的逆轉作用；其機製可能與下調凋亡抑製蛋白錶達相關.
목적 관찰붕체좌미대다약내약백혈병세포증식적영향급내약역전작용,초보탐토붕체좌미억제다약내약백혈세포증식적분자궤제.방법 이다약내약백혈병세포계HL-60/DNR화HL-60/VCR세포위모형,이HL-60세포위대조,용사갑기우담서염반응비색법(MTT법)측정붕체좌미대HL-60、HL-60/DNR화HL-60/VCR세포증식적억제작용,계산미독제량작위역전내약제량.분4개실험조:HL-60/DNR+유홍매소(DNR)、HL60/DNR+DNR+붕체좌미、HL-60/VCR+장춘신감(VCR)、HL-60/VCR +VCR+붕체좌미,계산붕체좌미역전내약배수.붕체좌미안농도체증(10、40、80nmol/L)작용우HL-60/DNR화HL-60/VCR세포48 h,실시정량RT-PCR화Western blot방법검측XIAP、cIAP-1、cIAP-2 mRNA화단백표체수평급NF-κB활성.결과 붕체좌미정농도의뢰성억제HL-60、HL-60/DNR화HL-60/VCR세포증식,IC50치분별위(28.90 ±3.99)、(81.19 ±9.34)화(73.48±8.94) nmol/L.10 nmol/L붕체좌미예처리48 h후,DNR대HL-60/DNR세포적IC50치유(12.90±1.75) μmol/L강저지(3.54±0.57) μmol/L(P ＜0.01),VCR대HL-60/VCR적IC50치유(33.25 ±7.28)μmol/L강저지(9.97±1.15) μmol/L(P ＜0.01),역전내약배수분별위(3.32±0.53)화(2.64±0.28)배.붕체좌미정농도의뢰성하조XIAP、cIAP-1、cIAP-2 mRNA화단백적표체수평급억제NF-κB활화.결론 붕체좌미가억제다약내약HL-60세포증식,병대다약내약구유일정적역전작용；기궤제가능여하조조망억제단백표체상관.
Objective To investigate the proliferation-inhibiting and multidrug-resistant reversing effect of bortezomib on human HL-60 cells,and to explore the mechanism of bortezomib-induced proliferation inhibition in human leukemia cells.Methods The multidrug resistant leukemia cell lines HL-60/DNR and HL-60/VCR cells were used as models,and sensitive HL-60 cells as a control.The cytotoxicity of bortezomib on HL-60,HL-60/DNR,HL-60/VCR cells were measured by MTT method,and the non-cytotoxicity dose was determined as reversible dose.The cells were divided into 4 experimental groups: HL-60/DNR + DNR,HL-60/DNR + DNR + bortezomib,HL-60/VCR + VCR,HL-60/VCR + VCR + bortezomib.The bortezomib resistant reversal fold was calculated.The levels of XIAP,cIAP-1,and cIAP-2 mRNA and proteins expression and the activation of NF-κB of the HL-60/DNR,HL-60/VCR cells were examined by quantitative real time RT-PCR and western blot respectively after treated with gradually increasing concentrations of bortezomib (10,40,80 nmol/L) for 48 hours.Results Bortezomib inhibited the cell growth of HL-60,HL-60/DNR,and HL-60/VCR in a concentration-dependent manner.The IC50 values were (28.90 ± 3.99),(81.19 ±9.34),and (73.48 ± 8.94) nmol/L,respectively.After treated with l0nmol/L bortezomib for 48 hours,the IC50 value of DNR to HL-60/DNR decreased from (12.90 ± 1.75) μmol/L to (3.54 ±0.57) μmol/L (P ＜0.01),and that of VCR to HL-60/VCR from (33.25 ± 7.28) μmol/L to (9.97 ± 1.15) μ mol/L (P ＜0.01).The reversal fold (RF) values were 3.32 ± 0.53 and 2.64 ± 0.28,respectively.Bortezomib downregulated the levels of XIAP,cIAP-1,and cIAP-2 mRNA and protein expression and inhibited the NF-κB activation in a concentration-dependent manner.Conclusion Bortezomib can inhibit the proliferation of HL-60 cells and reverse nultidrug-resistance in the cells.The possible mechanism is associated with downregulation of IAPs expression.
