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目的研究槲皮素(quercetin,Que)及山柰酚(kaempferol,Kae)单用及联合应用时对K562/A02细胞系多药耐药的影响及机制。方法体外培养K562和K562/A02细胞经Que及Kae处理,采用噻唑蓝(MTT)法计算药物单独及联合作用后细胞的生长抑制率及对多柔比星(adriamycin,ADM)的增敏倍数;流式细胞术检测药物作用后细胞内ADM浓度的变化;Annexin V/PI法观察细胞凋亡情况;Real-time PCR基因芯片检测药物转运蛋白及凋亡相关基因表达的变化。结果药物作用48 h后,Que及Kae在5～160μmol.L-1时对K562和K562/A02细胞均有剂量依赖性的生长抑制作用,K562/A02对ADM的敏感性显著增强,二者联用效果有协同性;在ADM为5μmol.L-1时,药物与细胞共培养2 h即可检出Que能增加细胞内ADM的浓度,而Kae则对ADM浓度无明显影响,两药联合作用的效果不及Que单独作用。Que和Kae均可剂量依赖性地诱导K562和K562/A02细胞的凋亡。二者可影响ABC、SLC家族等药物转运蛋白相关基因的表达,并可调节Bcl-2、TNF等凋亡相关基因的表达。结论 Que和Kae可通过多种途径逆转K562/A02的多药耐药性,但二者的作用机制可能不完全相同。
Objective To investigate the effects and mechanisms of quercetin (Que) and kaempferol (Kae) alone and in combination on multidrug resistance in K562 / A02 cell line. Methods K562 and K562 / A02 cells were treated with Que and Kae in vitro. The growth inhibitory rate and multiplication sensitivities of adriamycin (ADM) alone and in combination were measured by MTT assay. Flow cytometry was used to detect the changes of intracellular ADM concentration after the drug treatment. The apoptosis of cells was observed by Annexin V / PI assay. The expression of drug transporter and apoptosis related genes were detected by Real-time PCR. Results After 48 h treatment, both Que and Kae inhibited the growth of K562 and K562 / A02 cells at a dose of 5 ~ 160μmol.L-1. The sensitivity of K562 / A02 to ADM was significantly increased. Que could increase the intracellular ADM concentration when ADM was 5μmol·L-1 for 2 h, while Kae had no effect on the ADM concentration. The combination of the two drugs Less effective than Que alone. Both Que and Kae induced apoptosis in K562 and K562 / A02 cells in a dose-dependent manner. Both of them can influence the expression of drug transporter related genes such as ABC and SLC family, and regulate the expression of apoptosis-related genes such as Bcl-2 and TNF. Conclusions Que and Kae can reverse the multidrug resistance of K562 / A02 in a variety of ways, but the mechanism may not be the same.