姜黄素衍生物C1209对慢性粒细胞白血病细胞的作用及机制研究

Effects of Curcumin Derivative C1209 on CML Cells and Its Mechanism

目的研究姜黄素衍生物4-(4-甲酰基苯亚甲基)姜黄素(C1209)对慢性粒细胞白血病细胞凋亡及增殖的影响。方法MTT法检测C1209对白血病细胞K562及耐伊马替尼的白血病细胞K562/G_(01)的细胞毒性。流式细胞仪检测细胞线粒体膜电位、细胞凋亡及PI染色法测定细胞周期。Western blot法检测线粒体凋亡相关蛋白的表达。结果C1209作用24 h后,抑制K562和K562/G_(01)细胞的增殖,IC50为1.18和0.64μmol·L^(-1),且呈剂量依赖性。C1209破坏白血病细胞线粒体膜的完整性,引起线粒体膜电位下降,随着C1209浓度(0.4~3.2μmol·L^(-1))的增加,细胞凋亡率显著增高,并伴随着促凋亡蛋白Bax、Cleaved PARP、Cleaved Caspase-3/7/9上调,抗凋亡蛋白Bcl-2、PARP、Caspase-3/7/9下调或变化不明显。C1209阻滞细胞周期于S期。结论姜黄素衍生物C1209诱导细胞caspase途径介导的凋亡,抑制细胞增殖。

OBJECTIVE To study the effects of curcumin derivative C1209 on apoptosis and proliferation of chronic myeloid leukemia(CML)cells.METHODS MTT assay was used for determination of K562 and K562/G_(01) cells proliferation in vitro by C1209.By FITC/PI double staining the apoptotic effect of C1209 on K562 and K562/G_(01) cells was detected.The change of mitochondrial membrane potential of K562 and K562/G_(01) cells after C1209 treatment was observed by JC-1.PI staining was used for analysis of cell cycle.The effect of C1209 on mitochondrial apoptosis-related protein in CML was determined by Western blot.RESULTS The IC50 values of C1209 on K562 and K562/G_(01) cells were 1.18 and 0.64μmol·L^(-1) respectively after 24 h incubation.C1209 showed inhibitory effect on proliferation of K562 and K562/G_(01) cells in a dose-dependent manner.C1209 showed significant influence on the mitochondrial membrane potential of K562 and K562/G_(01) cells,increasing the mitochondrial membrane potential dissipation degree.In both imatinib-sensitive K562 chronic myeloid leukemia cells and imatinib-resistant K562/G_(01) chronic myeloid leukemia cells,C1209(0.4-3.2μmol·L^(-1))dose-dependently caused degradation or no obvious change of anti-apoptotic proteins(Bcl-2,PARP,Caspase-3/7/9)and up-regulated pro-apoptotic proteins(Bax,Cleaved PARP,Cleaved Caspase-3/7/9).Furthermore,C1209(0.4-3.2μmol·L^(-1))dose-dependently induced apoptosis of K562 and K562/G_(01) cells through triggering mitochondrial pathway.C1209 significantly blocked the cell cycle of K562 and K562/G_(01) cells in the S phase.CONCLUSION Curcumin derivative C1209 has the potential to exert its anti-leukemia effect through apoptosis induction of K562 and K562/G_(01) cells via activating caspase-9/caspase-3 mitochondrial pathway.