本课题研究槲皮素(quercetin,Que)对白血病细胞系K562/A多药耐药的影响及机制.体外培养的K562和K562/A细胞经不同浓度的Que处理,采用MTT法检测Que对细胞的生长抑制率及对阿霉紊(adriamycin,ADR)的增敏倍数;流式细胞术检测Que作用后细胞内ADR浓度的变化,Annexin V/PI双染色法观察细胞凋亡情况;实时荧光定量PCR基因芯片检测药物转运蛋白基因及凋亡相关基因表达的变化.结果表明,Que在5-160μmol/L的浓度范围内对K562和K562/A细胞均有剂量依赖性的生长抑制作用,低毒剂量的Que使K562/A对ADR的敏感性显著增强;在ADR为5 μmol/L时,Que与细胞共培养2小时细胞内ADR浓度则明显增加;Que可剂量依赖性地诱导K562和K562/A细胞的凋亡;Que可下调ABC、SLC家族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可通过多种机制逆转白血病细胞系K562/A的多药耐药性,逆转效果与剂量呈正相关.%The study was aimed to investigate the effect of quercetin, flavonoid molecules on reversing leukemia multidrug resistance and its mechanism. K562/A cells were cultured in vitro with different concentrations of quercetin.Cell growth inhibition and adriamycin (ADR) sensitivity were detected by MTT mathod. Intracellular ADR concentration was determined by flow cytometry. Cell apoptosis was assayed by Annexin V/PI staining method. The expressions of drug transporter and apoptosis related genes were measured by real-time PCR array. The results indicated that quercetin inhibited the proliferation of K562 and K562/A in 5 - 160 μmol/L and with dose-depenalent manner.Quercetin increased the sensitivity of K562/A cells to ADR in a low toxicity concentration. Flow cytometry showed that the quercetin increased the accumulation of ADR in K562/A cells when cells were co-cultured with 5 μnol/L ADR for 2 hours. Quercetin could induce the apoptosis of K562 and K562/A cells with dose dependent manner. Furthermore,some drug transport related genes such as ATP-binding cassette (ABC) and solute carrier (SLC) and some apoptosisrelated genes such as BCL-2, tumor necrosis factor ( TNF), rumor necrosis factor receptor (TNFR) families were downregulated by quercetin. It is concluded that quercetin reverses MDR of leukemic cells by multiple mechanisms and the reversing effect is positively related to drug concentration.
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