雷公藤甲素体外逆转急性髓系白血病耐药性  

Exploration of the ability of triptolide to reverse drug-resistance of adriamycin resistant acute myeloid leukemia cells

作　　者：王相朦[1] 陈菲莉[1] 柳约坚[1] 李蓉蔚[1] 王诗韵[1] 董慧娟[1] 韦祁[1] 周淑芸[1] 徐兵[1] 

机构地区：[1]南方医科大学南方医院血液科,广州510515

出　　处：《白血病．淋巴瘤》2013年第9期542-544,547,共4页Journal of Leukemia & Lymphoma

基　　金：国家自然科学基金（81070425）;广东省科技计划项目（201IB031800063）;南方医院院长基金（2012C007）

摘　　要：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（MTF）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；MTF法检测庀。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24h后缺氧诱导因子1α（HIF-1α）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24h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48h把。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48h IC20 TPL可使ADM对HL-60／ADM的IC20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。Westernblot结果显示TPL或ADM单药对HIF一10【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-1α和CXCR4蛋白的表达。结论IC20 TPL可逆转耐药AML细胞的耐药性，其分子机制与下调HIF-1α通路有关。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/adriamyein （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 MTF 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） p, mol/L, P 〈 0.05] to ADM. In comparison with antieancer agent alone, triptolide enhances the cytotoxicity of ADM [IC50：（14.36±2.23） μmol/L vs （7.90±0.33） μmol/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.

关 键 词：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 d亚基 

分 类 号：R733.71[医药卫生—肿瘤]