白血病·淋巴瘤
白血病·淋巴瘤
백혈병·림파류
JOURNAL OF LEUKEMIA & LYMPHOMA
2013年
9期
542-544,547
,共4页
王相朦%陈菲莉%柳约坚%李蓉蔚%王诗韵%董慧娟%韦祁%周淑芸%徐兵
王相朦%陳菲莉%柳約堅%李蓉蔚%王詩韻%董慧娟%韋祁%週淑蕓%徐兵
왕상몽%진비리%류약견%리용위%왕시운%동혜연%위기%주숙예%서병
雷公藤甲素%多柔比星%白血病,髓样,急性%细胞,HL-60%抗药性,肿瘤%缺氧诱导因子1,α亚基
雷公籐甲素%多柔比星%白血病,髓樣,急性%細胞,HL-60%抗藥性,腫瘤%缺氧誘導因子1,α亞基
뢰공등갑소%다유비성%백혈병,수양,급성%세포,HL-60%항약성,종류%결양유도인자1,α아기
Triptolide%Doxorubicin%Leukemia,myeloid,acute%Cells,HL-60%Drug resistance,neoplasm%Hypoxia-inducible factor 1,alpha subunit
目的 探讨小剂量雷公藤甲素(TPL)体外逆转急性髓系白血病(AML)耐药细胞的耐药性及其分子机制.方法 四甲基偶氮唑蓝(MTT)法检测多柔比星(ADM)对耐ADM的HL-60细胞株(HL-60/ADM)耐药情况及TPL对其增殖抑制作用；MTT法检测IC20浓度TPL联合不同浓度的ADM对HL-60/ADM的增殖抑制作用和两者之间的相互作用；Western blot检测IC20浓度TPL、ADM单药及两者联合作用HL-60/ADM细胞24 h后缺氧诱导因子1α(HIF-1 α)及CXC趋化因子受体4(CXCR4)蛋白表达水平的变化.结果 ADM对野生株HL-60细胞和ADM耐药细胞株24 h的IC50值分别为(0.28±0.35) μmol/L和(22.03±0.22)μmol/L,耐药细胞株的耐药倍数为78.68.TPL作用于HL-60/ADM细胞48 h IC50值为(43.06±1.06) nmol/L.对于HL-60/ADM细胞无明显增殖抑制作用的48 h IC20TPL可使ADM对HL-60/ADM的IC50值从(14.36±2.23) μmol/L降到(7.90±0.33) μmol/L,逆转耐药倍数为1.82倍,差异有统计学意义(P=0.008)；协同指数(CI)显示,在抑制率小于60％的情况下,ADM与TPL有协同作用.Western blot结果显示TPL或ADM单药对HIF-1α和CXCR4蛋白表达的下调作用均不明显,但TPL联合ADM可明显下调HIF-1 α和CXCR4蛋白的表达.结论 IC20TPL可逆转耐药AML细胞的耐药性,其分子机制与下调HIF-1 α通路有关.
目的 探討小劑量雷公籐甲素(TPL)體外逆轉急性髓繫白血病(AML)耐藥細胞的耐藥性及其分子機製.方法 四甲基偶氮唑藍(MTT)法檢測多柔比星(ADM)對耐ADM的HL-60細胞株(HL-60/ADM)耐藥情況及TPL對其增殖抑製作用；MTT法檢測IC20濃度TPL聯閤不同濃度的ADM對HL-60/ADM的增殖抑製作用和兩者之間的相互作用；Western blot檢測IC20濃度TPL、ADM單藥及兩者聯閤作用HL-60/ADM細胞24 h後缺氧誘導因子1α(HIF-1 α)及CXC趨化因子受體4(CXCR4)蛋白錶達水平的變化.結果 ADM對野生株HL-60細胞和ADM耐藥細胞株24 h的IC50值分彆為(0.28±0.35) μmol/L和(22.03±0.22)μmol/L,耐藥細胞株的耐藥倍數為78.68.TPL作用于HL-60/ADM細胞48 h IC50值為(43.06±1.06) nmol/L.對于HL-60/ADM細胞無明顯增殖抑製作用的48 h IC20TPL可使ADM對HL-60/ADM的IC50值從(14.36±2.23) μmol/L降到(7.90±0.33) μmol/L,逆轉耐藥倍數為1.82倍,差異有統計學意義(P=0.008)；協同指數(CI)顯示,在抑製率小于60％的情況下,ADM與TPL有協同作用.Western blot結果顯示TPL或ADM單藥對HIF-1α和CXCR4蛋白錶達的下調作用均不明顯,但TPL聯閤ADM可明顯下調HIF-1 α和CXCR4蛋白的錶達.結論 IC20TPL可逆轉耐藥AML細胞的耐藥性,其分子機製與下調HIF-1 α通路有關.
목적 탐토소제량뢰공등갑소(TPL)체외역전급성수계백혈병(AML)내약세포적내약성급기분자궤제.방법 사갑기우담서람(MTT)법검측다유비성(ADM)대내ADM적HL-60세포주(HL-60/ADM)내약정황급TPL대기증식억제작용；MTT법검측IC20농도TPL연합불동농도적ADM대HL-60/ADM적증식억제작용화량자지간적상호작용；Western blot검측IC20농도TPL、ADM단약급량자연합작용HL-60/ADM세포24 h후결양유도인자1α(HIF-1 α)급CXC추화인자수체4(CXCR4)단백표체수평적변화.결과 ADM대야생주HL-60세포화ADM내약세포주24 h적IC50치분별위(0.28±0.35) μmol/L화(22.03±0.22)μmol/L,내약세포주적내약배수위78.68.TPL작용우HL-60/ADM세포48 h IC50치위(43.06±1.06) nmol/L.대우HL-60/ADM세포무명현증식억제작용적48 h IC20TPL가사ADM대HL-60/ADM적IC50치종(14.36±2.23) μmol/L강도(7.90±0.33) μmol/L,역전내약배수위1.82배,차이유통계학의의(P=0.008)；협동지수(CI)현시,재억제솔소우60％적정황하,ADM여TPL유협동작용.Western blot결과현시TPL혹ADM단약대HIF-1α화CXCR4단백표체적하조작용균불명현,단TPL연합ADM가명현하조HIF-1 α화CXCR4단백적표체.결론 IC20TPL가역전내약AML세포적내약성,기분자궤제여하조HIF-1 α통로유관.
Objective Re-sensitization of leukemia resistant cell lines to common chemo-drug is the main method to achieve a better efficacy of treatment of acute myeloid leukemia (AML).This study uses cell line HL-60/adriamycin (ADM) which is resistant to ADM to evaluate whether low dose (IC20) of triptolide could reverse the resistance of HL-60/ADM to ADM and its mechanism.Methods HL-60/ADM cells were subjected to different treatments and thereafter MTT assay and Western blot or RT-PCR were used to determine the ability of triptolide to enhance the cytotoxicity of ADM to HL-60/ADM and expression of HIF-1α and their target genes.Results In comparison with the parental HL-60 cells [IC50 =(0.28±0.02) μmol/L],the HL-60/ADM cells were 78.68 folds resistant [IC50 =(22.03+0.22) μmol/L,P ＜ 0.05] to ADM.In comparison with anticancer agent alone,triptolide enhances the cytotoxicity of ADM [IC50:(14.36±2.23) μmol/L vs (7.90±0.33) μ mol/L,1.82 fold,P =0.008] to HL-60/ADM with a reverse fold being 1.82.Combination analysis showed the synergistic effects of triptolide and ADM when fraction affected was below 60 ％.Western blot analysis showed that expression of HIF-1α and CXCR4 were down-regulated in the largest scale in cells treated with tritolide combined with ADM.Conclusion Low-dose of triptolide could reverse the resistance of HL-60/ADM to ADM through down-regulation of HIF-1α pathway.
