Essential oils from Inula japonica and Angelicae dahuricae enhance sensitivity of MCF-7/ADR breast cancer cells to doxorubicin via multiple mechanisms
Essential oils from Inula japonica and Angelicae dahuricae enhance sensitivity of MCF-7/ADR breast cancer cells to doxorubicin via multiple mechanisms
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土木香和白芷精油通过多种机制增强 MCF-7/ADR 乳腺癌细胞对阿霉素的敏感性
DOI：
10.1016/j.jep.2016.01.015
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发表时间：
2016
影响因子：
5.4
通讯作者：
Liu Yiyao
中科院分区：
文献类型：
--
作者：
Wu Min;Li Tingting;Chen Lilan;Peng Sugang;Liao Wei;Bai Ruolan;Zhao Xue;Yang Hong;Wu Chunhui;Zeng Hongjuan;Liu Yiyao
民族药理相关性当归(Angelicae dahurica(Hoffm.)本斯。&Hook.f.ex Franch.葛根、天麻、旋覆花等中药配伍在中药配伍中应用广泛。以往的研究表明，当归挥发油(ADO)促进葛根素内化进入ABCB1过表达的Caco-2细胞。这些发现表明，精油可能通过与ABCB1相关的某些机制促进吸收，逆转多药耐药(MDR)。本研究目的：旋覆花精油(IJO)和旋覆花精油(IJO)可能逆转ABCB1介导的MDR，但这种能力尚未在成熟的癌细胞系中详细研究。本研究进一步探讨IJO和ADO逆转耐药人乳腺癌细胞株MCF-7/ADR多药耐药的分子机制。此外，本工作可能有助于揭示上述药物对ABCB1的配伍机制。材料和方法以非细胞毒性浓度的IJO及其倍半萜成分异丙内酯(ISO)或ADO处理MDR人乳腺癌MCF-7/ADR细胞。通过检测细胞对阿霉素(DOX)的敏感性、DOX蓄积和外排、ABCB1 ATPase活性、ABCB1表达、膜流动性以及脂筏和小窝的稳定性和定位来检测MDR能力。结果IJ油、ISO或AD油处理MCF-7/ADR细胞后，MDR逆转2~3倍，且不影响非MDR亲本细胞系的敏感性。机制研究表明，这些油下调ABCB1的mRNA和蛋白表达，并降低细胞膜中脂筏的稳定性，这已被证明减少了ABCB1介导的转运。另一方面，IJO、ISO和ADO不抑制ABCB1-ATPase的活性，荧光偏振实验表明，低浓度的油脂对膜流动性没有影响，与某些MDR逆转剂不同，ISO的对接分数高于维拉帕米，但低于多非奎达尔和替奎达尔。结论IJO、ISO和ADO通过下调ABCB1的表达，降低脂筏的稳定性，从而逆转MDR。这些发现可能有助于开发更安全有效的MDR逆转剂，也有助于了解其配伍机制。
Ethnopharmacological relevanceAngelicae dahurica (Hoffm.) Benth. & Hook.f.ex Franch. & Savcombined withPuerariaandGastrodia elataBl.combined withInulajaponica Thunb.are widely used in herb-pairs of traditional chinese medicine. Previous studies have shown thatAngelicae dahuricaeessential oil (ADO) enhanced puerarin internalization into ABCB1-overexpressed Caco-2 cells. These findings suggest the possibility that essential oils may enhance the absorption via certain mechanisms related to ABCB1 and reverse multidrug resistance (MDR).Aim of the studyADO and essential oils fromInula japonica(IJO) may reverse ABCB1-mediated MDR, but this ability has not been investigated in detail in the well-established cancer cell lines. In this study, the underlying molecular mechanisms were further investigated to examine how IJO and ADO reverse MDR in the resistant human breast cancer cell line of MCF-7/ADR. Also this work may help uncover the conceivable compatibility mechanisms of above herb-pairs involved in ABCB1.Materials and methodsThe MDR human breast cancer MCF-7/ADR cells were treated with IJO, its sesquiterpene component isoalantolactone (ISO) or ADOat non- cytotoxic concentrations. The MDR ability was examined by measuring the sensitivity to doxorubicin (DOX), DOX accumulation and efflux, ABCB1 ATPase activity, ABCB1 expression, membrane fluidity, and stability and localization of lipid rafts and caveolae. Finally, the molecular modeling was performed to postulate how ISO interacts with ABCB1.ResultsTreating MCF-7/ADR cells with IJ oil, ISO or AD oil reversed MDR 2- to 3-fold, without affecting the sensitivity of the non-MDR parental cell line. Mechanistic studies showed that these oils down-regulated mRNA and protein expression of ABCB1, and reduced the stability of lipid rafts in the cell membrane, which has previously been shown to reduce ABCB1-mediated transport. On the other hand, IJO, ISO and ADO did not inhibit ABCB1 ATPase activity, and fluorescence polarization experiments showed that low concentrations of the oils did not appear to alter membrane fluidity, unlike some MDR-reversing agents, ISO showed a higher docking score than verapamil but lower than dofequidar and tariquidar.ConclusionsOur results suggest that IJO, ISO and ADO could reverse MDR by down-regulating ABCB1 expression and reducing lipid raft stability. These findings may be useful for developing safer and effective MDR reversal agents and also help find out the compatibility mechanisms.