Chemosensitization of hepatocellular carcinoma cells to sorafenib by β-caryophyllene oxide-induced inhibition of ABC export pumps
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Several ATP-binding cassette (ABC) proteins reduce intracellular concentrations of antitumor drugs and hence weaken the response of cancer cells to chemotherapy. Accordingly, the inhibition of these export pumps constitutes a promising strategy to chemosensitize highly chemoresistant tumors, such as hepatocellular carcinoma (HCC). Here, we have investigated the ability of β-caryophyllene oxide (CRYO), a naturally occurring sesquiterpene component of many essential oils, to inhibit, at non-toxic doses, ABC pumps and improve the response of HCC cells to sorafenib. First, we have obtained a clonal subline (Alexander/R) derived from human hepatoma cells with enhanced multidrug resistance (MDR) associated to up-regulation (mRNA and protein) of MRP1 and MRP2. Analysis of fluorescent substrates export (flow cytometry) revealed that CRYO did not affect the efflux of fluorescein (MRP3, MRP4 and MRP5) but inhibited that of rhodamine 123 (MDR1) and calcein (MRP1 and MRP2). This ability was higher for CRYO than for other sesquiterpenes assayed. CRYO also inhibited sorafenib efflux, increased its intracellular accumulation (HPLC–MS/MS) and enhanced its cytotoxic response (MTT). For comparison, the effect of known ABC pumps inhibitors was also determined. They induced strong (diclofenac on MRPs), modest (verapamil on MDR1) or null (fumitremorgin C on BCRP) effect on sorafenib efflux and cytotoxicity. In the mouse xenograft model, the response to sorafenib treatment of subcutaneous tumors generated by mouse hepatoma Hepa 1–6/R cells, with marked MDR phenotype, was significantly enhanced by CRYO co-administration. In conclusion, at non-toxic dose, CRYO is able to chemosensitizating liver cancer cells to sorafenib by favoring its intracellular accumulation.
KeywordsChemotherapy Liver cancer Multidrug resistance Sesquiterpenes Tyrosine kinase inhibitor
This study was supported by the CIBERehd (EHD15PI05/2016) and Fondo de Investigaciones Sanitarias, Instituto de Salud Carlos III, Spain (PI16/00598, co-funded by European Regional Development Fund/European Social Fund, “Investing in your future”); Spanish Ministry of Economy, Industry and Competitiveness (SAF2016-75197-R); Junta de Castilla y Leon (SA063P17); Fundación Mutua Madrileña, Spain (Call 2015); Fundación Samuel Solórzano Barruso, Spain (FS/7-2016, FS/8-2017 and FS/13-2017); AECC Scientific Foundation (2017/2020), Spain. S. Di Giacomo and A. Di Sotto were supported by the “Enrico and Enrica Sovena” Foundation (Italy); S. Di Giacomo was also financed by the Italian Society of Pharmacology (SIF).
Compliance with ethical standards
Conflict of interest
The authors declare that there is no conflict of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were in accordance with the ethical standards of the institution or practice at which the studies were conducted.
- Ambroz M, Bousova I, Skarka A et al (2015) The influence of sesquiterpenes from Myrica rubra on the antiproliferative and pro-oxidative effects of doxorubicin and its accumulation in cancer cells. Molecules 20(8):15343–15358. https://doi.org/10.3390/molecules200815343 CrossRefPubMedPubMedCentralGoogle Scholar
- Brozik A, Hegedus C, Erdei Z et al (2011) Tyrosine kinase inhibitors as modulators of ATP binding cassette multidrug transporters: substrates, chemosensitizers or inducers of acquired multidrug resistance? Expert Opin Drug Metab Toxicol 7(5):623–642. https://doi.org/10.1517/17425255.2011.562892 CrossRefPubMedGoogle Scholar
- Munoz-Martinez F, Lu P, Cortes-Selva F et al (2004) Celastraceae sesquiterpenes as a new class of modulators that bind specifically to human P-glycoprotein and reverse cellular multidrug resistance. Cancer Res 64(19):7130–7138. https://doi.org/10.1158/0008-5472.CAN-04-1005 CrossRefPubMedGoogle Scholar
- Oliveira-Tintino CDM, Pessoa RT, Fernandes MNM et al (2018) Anti-inflammatory and anti-edematogenic action of the Croton campestris A. St.-Hil (Euphorbiaceae) essential oil and the compound beta-caryophyllene in in vivo models. Phytomedicine 41:82–95. https://doi.org/10.1016/j.phymed.2018.02.004 CrossRefPubMedGoogle Scholar
- Vander Borght S, Komuta M, Libbrecht L et al (2008) Expression of multidrug resistance-associated protein 1 in hepatocellular carcinoma is associated with a more aggressive tumour phenotype and may reflect a progenitor cell origin. Liver Int 28(10):1370–1380. https://doi.org/10.1111/j.1478-3231.2008.01889.x CrossRefPubMedGoogle Scholar
- Wang H, Jia XH, Chen JR et al (2016) HOXB4 knockdown reverses multidrug resistance of human myelogenous leukemia K562/ADM cells by downregulating P-gp, MRP1 and BCRP expression via PI3K/Akt signaling pathway. Int J Oncol 49(6):2529–2537. https://doi.org/10.3892/ijo.2016.3738 CrossRefPubMedGoogle Scholar