长春西汀通过抑制 PI3K/Nrf2 刺激的 MCF-7 和 K562 DOX 耐药细胞系中 MRP1 的表达增强多柔比星的细胞毒性。
Vielanin P enhances the cytotoxicity of doxorubicin via the inhibition of PI3K/Nrf2-stimulated MRP1 expression in MCF-7 and K562 DOX-resistant cell lines.
机构信息
Jiangsu Key Laboratory of Bioactive Natural Product Research and State Key Laboratory of Natural Medicines, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, China.
Jiangsu Key Laboratory of Bioactive Natural Product Research and State Key Laboratory of Natural Medicines, China Pharmaceutical University, 24 Tong Jia Xiang, Nanjing 210009, China.
出版信息
Phytomedicine. 2019 May;58:152885. doi: 10.1016/j.phymed.2019.152885. Epub 2019 Mar 11.
BACKGROUND
Cancer cells that are resistant to structurally and mechanically unrelated anticancer drugs are said to have multidrug resistance (MDR). The overexpression of the ATP-binding cassette (ABC) transporter is one of the most important mechanisms of MDR. Vielanin P (VP), a dimeric guaiane from the leaves of Xylopia vielana, has the potential to reverse multidrug resistance.
PURPOSE
To evaluate the meroterpenoid compound VP as a low cytotoxicity MDR regulator and the related mechanisms.
METHODS
Cell viability was determined by CCK-8 and MTT assays. Apoptosis and the accumulation of doxorubicin (DOX) and 5(6)-carboxyfluorescein diacetate (CFDA) were determined by flow cytometry. We determined mRNA levels by quantitative real-time polymerase chain reaction (qRT-PCR). Protein levels were analyzed by Western blotting and immunofluorescence.
RESULTS
In the MCF-7 and K562 DOX-resistant cell lines, VP treatment (10 μM or 20 μM) enhanced the activity of chemotherapeutic agents. We found that VP selectively inhibited MRP1 mRNA but not MDR1 mRNA. VP enhanced DOX-induced apoptosis and reduced colony formation in the presence of DOX in drug-resistant cells. Moreover, VP increased the accumulation of DOX and the MRP1-specific substrate CFDA. In addition, VP reversed MRP1 protein levels and the accumulation of DOX and CFDA in MRP1-overexpressing MCF-7 and K562 cells. Thus, the mechanism of MDR reversal by VP is MRP1-dependent. Furthermore, we found that the inhibitory effect of VP on MRP1 is PI3K/Nrf2-dependent.
CONCLUSION
These results support the potential therapeutic value of VP as an MDR-reversal agent by inhibiting MRP1 via PI3K/Nrf2 signaling.
背景
对结构和机械上不相关的抗癌药物具有耐药性的癌细胞被称为多药耐药（MDR）。ATP 结合盒（ABC）转运蛋白的过度表达是 MDR 的最重要机制之一。Vielanin P（VP）是来自 Xylopia vielana 叶子的二聚古烷，具有逆转多药耐药的潜力。
目的
评估倍半萜化合物 VP 作为低细胞毒性 MDR 调节剂及其相关机制。
方法
通过 CCK-8 和 MTT 测定法测定细胞活力。通过流式细胞术测定阿霉素（DOX）和 5（6）-羧基荧光素二乙酸酯（CFDA）的积累和细胞凋亡。通过定量实时聚合酶链反应（qRT-PCR）测定 mRNA 水平。通过 Western 印迹和免疫荧光分析蛋白质水平。
结果
在 MCF-7 和 K562 DOX 耐药细胞系中，VP 处理（10μM 或 20μM）增强了化疗药物的活性。我们发现 VP 选择性抑制 MRP1 mRNA，但不抑制 MDR1 mRNA。VP 增强了 DOX 诱导的细胞凋亡，并减少了耐药细胞中 DOX 存在时的集落形成。此外，VP 增加了 DOX 和 MRP1 特异性底物 CFDA 的积累。此外，VP 逆转了 MRP1 过表达的 MCF-7 和 K562 细胞中 MRP1 蛋白水平和 DOX 和 CFDA 的积累。因此，VP 逆转 MDR 的机制是 MRP1 依赖性的。此外，我们发现 VP 对 MRP1 的抑制作用依赖于 PI3K/Nrf2。
结论
这些结果支持 VP 通过 PI3K/Nrf2 信号抑制 MRP1 作为 MDR 逆转剂的潜在治疗价值。
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