目的 探索木香烴內酯對白血病耐藥細胞系K562/A02阿霉素耐藥的逆轉作用。方法 將木香烴內酯與K562/A02細胞共培養(yǎng)72 h后,采用MTT法檢測木香烴內酯對細胞生長的抑制作用。不同濃度木香烴內酯處理K562/A02細胞24 h后,采用Annexin V和PI雙染法檢測木香烴內酯對細胞的凋亡。不同濃度木香烴內酯與阿霉素共培養(yǎng)72 h后采用MTT法檢測木香烴內酯對K562/A02細胞的耐藥逆轉情況。采用流式細胞儀檢測木香烴內酯處理24 h之后K562/A02細胞阿霉素蓄積以及細胞膜表面P-糖蛋白(P-gp)的表達。結果 木香烴內酯對K562/A02細胞的生長具有明顯抑制作用,呈顯著的劑量相關性。與對照組比較,2.5~50 μmol/L木香烴內酯組K562/A02細胞存活率顯著降低(P<0.05、0.01、0.001)。隨著木香烴內酯濃度的增加,凋亡細胞的比例明顯增加。與對照組比較,不同濃度木香烴內酯組細胞凋亡比例顯著升高(P<0.05、0.01、0.001)。加入5 μmol/L木香烴內酯后,使K562/A02細胞對阿霉素的敏感性提高12倍。木香烴內酯處理后K562/A02細胞內部阿霉素的蓄積明顯增加,呈濃度相關性。與對照組比較,5、10 μmol/L木香烴內酯組K562/A02細胞內部阿霉素的蓄積增加顯著升高(P<0.05)。細胞表面的P-gp的表達并無顯著影響。結論 木香烴內酯能夠抑制K562/A02細胞增殖,誘導K562/A02細胞凋亡,增強阿霉素的化療敏感性,逆轉阿霉素耐藥。

Objective To investigate the reverse effect against adriamycin resistance of costunolide on leukemia drug resistant cell line K562/A02. Methods Costunolide and K562/A02 cells were co-cultured for 72 h, and inhibition of costunolide on K562/A02 cell were determined by MTT assay. K562/A02 cells were treated for 24 h by various concentrations of costunolide, and the apoptosis of K562/A02 cells were detected by Annexin V/PI apoptosis kit on flow cytometry. Various concentrations of costunolide and adriamycin were co-cultured with K562/A02 cells for 72 h, and the reversals were determined by MTT assay. Accumulation of adriamycin and the expression of P-glycoprotein (P-gp) were analyzed by flow cytometry after the treatment for 24 h of costunolide with various concentrations. Results Costunolide could significantly exhibit growth of K562/A02 cell in a dose dependent manner. Compared with the control group, survival rates of K562/A02 cell in the costunolide 2.5 - 50 μmol/L groups were significantly decreased (P < 0.05, 0.01, 0.001). Apoptosis ratios of K562/A02 cells in the costunolide groups were significantly decreased with increase of concentrations of costunolide. Compared with the control group, apoptosis rates of K562/A02 cell in the costunolide groups were significantly increased (P < 0.05, 0.01, 0.001). Intake of adriamycin in K562/A02 cell increased to 12 times after treated by 5 μmol/L costunolide. Accumulation of adriamycin in K562/A02 cell treated by costunolide were significantly increased in a concentration dependent manner. Compared with the control group, accumulation of adriamycin in K562/A02 cell in the 5 and 10 μmol/L costunolide groups were significantly increased (P < 0.05). However, there was no significant difference of expressions of P-gp between two groups. Conclusion Costunolide can inhibit the proliferation of K562/A02 cells, induce apoptosis in K562/A02 cells, enhance chemotherapy sensitivity of doxorubicin, and reverse adriamycin resistance.

國家自然科學基金資助項目(81570100)