Chen Cheng, Wang Ji-Shi, Ynag Chang, Yu Yan-Yan, Li Yan, Ma Dan, Fang Qin
Department of Hematology, Guiyang Medical College, Guiyang, Guizhou Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2012 Aug;20(4):867-71.
This study was aimed to investigate the effect of AMN107 (nilotinib) combined with heme oxygenase-1 (HO-1) inhibitor zinc protoporphyrin IX (ZnPPIX) on chronic myeloid leukemia (CML) cells and its mechanism. Proliferative 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, ZnPPIX group, AMN107 group, AMN107 combined with ZnPPIX group at 48 h respectively. It is concluded that the second-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.
本研究旨在探讨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）对慢性髓性白血病（CML）细胞的影响及其机制。采用MTT法和台盼蓝法观察单独或联合使用AMN107（10µmol/L）和ZnPPIX（10µmol/L）处理不同时间的细胞增殖率；于48h时，通过半定量RT-PCR和Western blot检测对照组、ZnPPIX（10µmol/L）组、AMN107（10µmol/L）组、AMN107（10µmol/L）联合ZnPPIX（10µmol/L）组中HO-1的表达。48h时，采用Annexin V/PI双染法通过流式细胞术检测细胞凋亡。结果显示，联合组对细胞增殖的抑制作用最强，且呈时间依赖性；联合组中HO-1的表达水平最低；48h时，对照组、ZnPPIX组、AMN107组、AMN107联合ZnPPIX组的细胞凋亡率分别为（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可诱导CML细胞凋亡。抑制HO-1表达可增强AMN107对CML细胞的杀伤作用，为进一步提高CML治疗的临床疗效提供了实验依据。
