目的 探讨具有较好水解稳定性的姜黄素结构类似物的抗多药耐药肿瘤活性及机制。方法 对人白血病多药耐药细胞K562/A02孵育姜黄素及其类似物1～19，MTT法筛选其对K562/A02细胞的生长抑制作用；高效液相色谱法测定姜黄素及其类似物3、4的水解稳定性；流式细胞术检测姜黄素及其类似物3、4对K562/A02细胞凋亡、线粒体膜电位（MMP）、活性氧（ROS）量及细胞周期分布的影响；分光光度法检测K562/A02细胞caspase-3活性变化；Western blotting法检测K562/A02细胞P-糖蛋白（P-gp）表达、荧光法检测K562/A02细胞内阿霉素的累积效应。结果 姜黄素结构类似物3、4与姜黄素对K562/A02细胞的生长抑制作用相近，但类似物3、4水解稳定性较姜黄素有显著增高；与对照组比较，姜黄素及其类似物3、4均能够通过降低K562/A02细胞MMP、增加caspase-3活性促进细胞凋亡，但此过程中细胞内ROS并未显著增加；姜黄素及其类似物3、4可显著降低K562/A02细胞G₂/M期和S期的细胞比例，亚二倍体增加；Western blotting结果显示姜黄素及其类似物3、4对P-gp有显著下调作用，可增加细胞内阿霉素的累积。结论 姜黄素结构类似物3、4具有较高的水解稳定性，且保持了姜黄素的抗多药耐药肿瘤活性。

Objective To investigate the effect and mechanisms of a series of curcumin analogues with better hydrolytic stability on multidrug-resistant tumor. Methods Human leukemia multi-drug resistant K562/A02 cells were incubated with curcumin and its analogues 1－19. MTT method was applied for screening the inhibition on the growth of K562/A02 cells; HPLC was used to detect the hydrolytic stability of curcumin and its analogues 3 and 4; The effects of curcumin and its analogues 3 and 4 on apoptosis, mitochondrial membrane potential (MMP), reaction oxygen species (ROS) content, and cell cycle distribution of K562/A02 cells were measured by flow cytometry; Spectrophotometric method was used to detect the caspase-3 activity; The expression of P-glycoprotein (P-gp) was assayed by Western blotting; Fluorometric method was applied to evaluating the intracellular Adriamycin accumulation of anti-tumor drug. Results Analogues 3 and 4 exhibited the inhibitory effect on the growth of K562/A02 cells as similar as that of curcumin, but their hydrolytic stability was significantly higher than that of curcumin. Compared with the control group, curcumin and its analogues 3 and 4 could significantly promote apoptosis by reducing MMP and increasing caspase-3 activity of K562/A02 cells, but no obvious increase in intracellular ROS was found. In addition, the cell proportion of K562/A02 cells in G₂/M and S phases was significantly reduced in the presence of curcumin and its analogues, while the hypodiploid increased. Besides, Western blotting indicated that the P-gp was significantly downregulated with an increase of intracellular anti-tumor drug accumulation. Conclusion The analogues 3 and 4 of curcumin have the better hydrolytic stability than that of curcumin, while the anti-resistant tumor activity is maintained.

国家自然科学基金资助项目（U1204830）；教育部留学归国人员科研启动基金；河南省教育厅科学技术研究重点项目资助计划项目（13A310064）