AMN107联合血红素加氧酶-1抑制剂诱导慢性髓系白血病细胞凋亡  

CML Cell Apotosis Induced by AMN107 Combined with Heme Oxygenase-1 Inhibitor

作　　者：陈珵[1] 王季石[1] 杨畅[2] 于艳艳[1] 李燕[1] 马丹[2] 方琴[2] 

机构地区：[1]贵阳医学院附属医院血液科,贵州贵阳550004 [2]贵阳医学院附属医院药剂科,贵州贵阳550004

出　　处：《中国实验血液学杂志》2012年第4期867-871,共5页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(编号30760276;30460127;81070444);黔科平台[编号2009]4007;贵州省科技厅基金(编号E2010-6);贵阳市科技局基金(编号T2009-5)

摘　　要：运用AMN107(尼洛替尼)联合血红素加氧酶-1(HO-1)抑制剂锌原卟啉Ⅸ(ZnPPⅨ)作用于慢性髓系白血病(CML)细胞株K562细胞,研究其对CML细胞增殖的影响并探讨其作用机制。采用MTT法和台盼蓝染色法检测AMN107(10μmol/L)及ZnPPⅨ(10μmol/L)单独或联合处理不同时间的细胞增殖率;半定量RT-PCR法和Westernblot法检测空白对照组、ZnPPⅨ(10μmol/L)组、AMN107(10μmol/L)组、AMN107(10μmol/L)联合ZnPPⅨ(10μmol/L)组48 h时细胞HO-1的表达;AnnexinⅤ/PI双染色法检测处理48 h时各组细胞凋亡情况。结果表明:联合用药对细胞的抑制作用最强,且呈时间依赖性;联合用药组HO-1的表达量最低;空白对照组、ZnPPⅨ(10μmol/L)组、AMN107(10μmol/L)组、AMN107(10μmol/L)联合ZnPPⅨ(10μmol/L)组48 h时细胞凋亡率分别为(11.38±0.02)%、(17.44±0.08)%、(39.81±0.07)%和(56.46±0.19)%。结论:第二代酪氨酸激酶抑制剂AMN107具有诱导CML细胞凋亡的作用;抑制HO-1表达能加强AMN107对CML细胞的杀伤作用,这为临床进一步提高CML的疗效提供实验依据。This study was aimed to investigate the effect of AMN107 （nilotinib） combined with heine oxygenase-1 （HO-1） inhibitor zinc protoporphyrinlX （ZnPPⅨ） on chronic myeloid leukemia （CML） cells and its mechanism. Pro- liferative rate of cells treated with AMN107（ 10 μmol/L）and ZnPPⅨ （10 μmol/L）alone or both for different time was observed by MTT and trypan blue methods; the expression of HO-1 in the control group, ZnPPⅨ （10 μmol/L） group, AMN107（ 10 μmol/L） group, AMN107 （ 10 μmol/L） combined with ZnPPⅨ （ 10 μmol/L） group was evaluated by semi-quantitative RT-PCR and Western blot at 48 h. Cell apoptosis was detected by flow cytometry with Annexin V/PI double staining at 48 h. The results showed that the strongest inhibition of cell proliferation was detected in combined group, and in a time-dependent manner; the expression level of HO-1 was lowest in combined group; the cell apoptosis rates were （11.38 ±0.02）%, （17.44 ±0.08）%, （39.81 ±0.07）% and （56.46 ±0.19）% in the control group, Zn- PPⅨ group, AMN107 group, AMN107 combined with ZnPPⅨ group at 48 h respectively. It is concluded that the sec- ond-generation tyrosine kinase inhibitor AMN107 can induce the apoptosis in CML cells. Inhibition of HO-1 expression can enhance the killing effect of AMN107 on CML cells, which provides experimental evidence to further improve the clinical efficacy of CML treatment.

关 键 词：AMN107 锌原卟啉Ⅸ 血红素加氧酶-1 K562细胞 慢性髓系白血病 细胞凋亡 

分 类 号：R733.72[医药卫生—肿瘤] R979.1[医药卫生—临床医学]