姜黄素抑制多药耐药肝癌细胞HepG2/ADM的增殖及其机制研究

摘要

目的 观察姜黄素对多药耐药(MDR)的肝癌细胞株HepG2/ADM细胞增殖的抑制作用及其机制.方法 制备、培养HepG2/ADM细胞,然后用不同浓度的姜黄素(5、10、20、40 mol/L)处理该细胞24、48、72 h.采用CCK-8试剂检测姜黄索对HepG2/ADM细胞的增殖活力的影响.流式细胞仪检测细胞内罗丹明123(R-123)的浓度和阿霉素(ADM)的浓度;反转录-聚合酶链反应(RT-PCR)检测各组细胞内mdr-1 mRNA的水平变化;用Western blot检测细胞内p-糖蛋白(pgp)水平的变化.结果 与空白对照组和DMSO组相比,姜黄素对HepG2/ADM细胞增殖的抑制作用更加明显(P＜0.05);更能够明显抑制细胞内Rh123的外排(P＜0.05);RT-PCR和Western blot结果分别显示,姜黄素对HepG2/ADM细胞内的mdr-1 mRNA和P-gp水平更有明显的降低(P＜0.05),且呈浓度-时间依赖性(P＜0.05).结论 姜黄素可以显著抑制多药耐药HepG2/ADM细胞的增殖,其机制可能与抑制和MDR密切相关的mdr-1基因及其编码的P-gp水平有关.%Objective To observe the inhibitory effect of curcumin on the proliferation of multidrug resistance liver cancer line HepG2/ADM cells and to explore its mechanisms.Methods HepG2/ADM cells were prepared and cultured in vitro,and treated by different concentrations (5,10,20,40 μmol/L) of curcumin for 24,48,72 h respectively.The effect of curcumin on proliferation of HepG2/ADM cells was measured by CCK-8 reagent;the concentration of intracellular rhodamine-123 (Rh-123) and adriamycin (ADM) were determined by flow cytometry;the level change of intracellular mdr-1 mRNA in each group was determined by RT-PCR,the P-gp protein level was detected by Western blot.Results Compared with the blank control and DMSO group,curucmin had more obvious inhibitory effect on HepG2/ADM cells proliferation (P＜0.05),and could more remarkably inhibit the intracellular Rh-123 excretion(P＜0.05).The RT-PCR and Western blot results showed that curcumm more significantly decrease the mdr-1 mRNA and P-gp protein levels in dose-time dependent manner (P＜0.05).Conclusion Curcumin could significantly inhibit the proliferation of multidrug-resistant HepG2/ADM cells,and its mechanism may be related with inhibiting mdr-1 gene expression and its encoded P-gp protein level,which are closely related with MDR.