黄酮类化合物通过 Wnt 信号通路调节 P-糖蛋白过度表达细胞系中的多药耐药性。
Flavonoids modulate multidrug resistance through wnt signaling in P-glycoprotein overexpressing cell lines.
机构信息
Department of Biochemistry and Biotechnology, Annamalai University, Annamalai Nagar, Tamil Nadu, 608 002, India.
Bioinformatics Infrastructure Facility (BIF), University of Madras, Guindy Campus, Chennai, Tamil Nadu, India.
出版信息
BMC Cancer. 2018 Nov 26;18(1):1168. doi: 10.1186/s12885-018-5103-1.
BACKGROUND
Wnt signaling has been linked with P-glycoprotein (P-gp) overexpression and which was mainly mediated by β-catenin nuclear translocation. Flavonoids have already been reported as modulators of the Wnt/β-catenin pathway and hence they may serve as promising agents in the reversal of P-gp mediated cancer multi drug resistance (MDR).
METHODS
In this study, we screened selected flavonoids against Wnt/β-catenin signaling molecules. The binding interaction of flavonoids (theaflavin, quercetin, rutin, epicatechin 3 gallate and tamarixetin) with GSK 3β was determined by molecular docking. Flavonoids on P-gp expression and the components of Wnt signaling in drug-resistant KBCH8-5 cells were analyzed by western blotting and qRT-PCR. The MDR reversal potential of these selected flavonoids against P-gp mediated drug resistance was analyzed by cytotoxicity assay in KBCH8-5 and MCF7/ADR cell lines. The chemosensitizing potential of flavonoids was further analyzed by observing cell cycle arrest in KBCH8-5 cells.
RESULTS
In this study, we observed that the components of Wnt/β-catenin pathway such as Wnt and GSK 3β were activated in multidrug resistant KBCH8-5 cell lines. All the flavonoids selected in this study significantly decreased the expression of Wnt and GSK 3β in KBCH8-5 cells and subsequently modulates P-gp overexpression in this drug-resistant cell line. Further, we observed that these flavonoids considerably decreased the doxorubicin resistance in KBCH8-5 and MCF7/ADR cell lines. The MDR reversal potential of flavonoids were found to be in the order of theaflavin > quercetin > rutin > epicatechin 3 gallate > tamarixetin. Moreover, we observed that flavonoids pretreatment significantly induced the doxorubicin-mediated arrest at the phase of G2/M. Further, the combinations of doxorubicin with flavonoids significantly modulate the expression of drug response genes in KBCH8-5 cells.
CONCLUSION
The present findings illustrate that the studied flavonoids significantly enhances doxorubicin-mediated cell death through modulating P-gp expression pattern by targeting Wnt/β-catenin signaling in drug-resistant KBCH8-5 cells.
背景
Wnt 信号与 P-糖蛋白（P-gp）过表达有关，主要由β-连环蛋白核转位介导。黄酮类化合物已被报道为 Wnt/β-连环蛋白通路的调节剂，因此它们可能成为逆转 P-gp 介导的癌症多药耐药（MDR）的有前途的药物。
方法
在这项研究中，我们筛选了几种黄酮类化合物来对抗 Wnt/β-连环蛋白信号分子。通过分子对接确定黄酮类化合物（茶黄素、槲皮素、芦丁、表儿茶素 3 没食子酸酯和柽柳素）与 GSK 3β 的结合相互作用。用 Western blot 和 qRT-PCR 分析黄酮类化合物对耐药 KBCH8-5 细胞中 P-gp 表达和 Wnt 信号成分的影响。用细胞毒性测定法在 KBCH8-5 和 MCF7/ADR 细胞系中分析这些选定的黄酮类化合物对 P-gp 介导的耐药性的逆转潜力。通过观察 KBCH8-5 细胞中的细胞周期停滞进一步分析黄酮类化合物的化疗增敏作用。
结果
在这项研究中，我们观察到多药耐药 KBCH8-5 细胞系中 Wnt/β-连环蛋白通路的成分如 Wnt 和 GSK 3β 被激活。本研究中选择的所有黄酮类化合物均显著降低了 KBCH8-5 细胞中 Wnt 和 GSK 3β 的表达，并随后调节了该耐药细胞系中 P-gp 的过度表达。此外，我们观察到这些黄酮类化合物显著降低了 KBCH8-5 和 MCF7/ADR 细胞系中多柔比星的耐药性。黄酮类化合物的 MDR 逆转潜力顺序为茶黄素＞槲皮素＞芦丁＞表儿茶素 3 没食子酸酯＞柽柳素。此外，我们观察到黄酮类化合物预处理可显著诱导多柔比星在 G2/M 期的阻滞。此外，多柔比星与黄酮类化合物的联合使用可显著调节 KBCH8-5 细胞中药物反应基因的表达。
结论
本研究结果表明，在所研究的黄酮类化合物通过靶向耐药 KBCH8-5 细胞中的 Wnt/β-连环蛋白信号来调节 P-gp 表达模式，显著增强了多柔比星介导的细胞死亡。
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