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Cancer Chemotherapy and Pharmacology

, Volume 76, Issue 1, pp 105–116 | Cite as

Interactions of cyclin-dependent kinase inhibitors AT-7519, flavopiridol and SNS-032 with ABCB1, ABCG2 and ABCC1 transporters and their potential to overcome multidrug resistance in vitro

  • Daniela Cihalova
  • Frantisek Staud
  • Martina Ceckova
Original Article

Abstract

Purpose

ATP-binding cassette (ABC) transporters play an important role in multidrug resistance (MDR) toward anticancer drugs. Here, we evaluated interactions of cyclin-dependent kinase inhibitors (CDKi) AT-7519, flavopiridol and SNS-032 with the following ABC transporters in vitro: P-glycoprotein (ABCB1), breast cancer resistance protein (ABCG2) and multidrug resistance-associated protein 1 (ABCC1).

Methods

Inhibitory potency of studied CDKi to the transporters was evaluated by accumulation assays using fluorescent substrates and MDCKII cells overexpressing human ABCB1, ABCG2 or ABCC1. Resistance of transporter-expressing cells to the CDKi was evaluated by XTT proliferation assay. Observed interactions of CDKi were verified by ATPase assay in ABC transporter-expressing Sf9 membrane vesicles. Combination index analysis was additionally performed in ABC transporter-expressing cancer cell lines, HepG2 and T47D.

Results

Flavopiridol showed a significant inhibitory potency toward ABCG2 and ABCC1. SNS-032 also decreased ABCG2-mediated efflux, while AT-7519 failed to inhibit ABCB1, ABCG2 or ABCC1. Both flavopiridol and SNS-032 showed synergistic antiproliferative effects in combination with relevant ABC transporter substrates such as daunorubicin and topotecan in cancer cells. ABCB1 was found to confer significant resistance to AT-7519 and SNS-032, but not to flavopiridol. In contrast, ABCG2 and ABCC1 conferred resistance to flavopiridol, but not to AT-7519 and SNS-032.

Conclusion

Our data provide detailed information on interactions of flavopiridol, SNS-032 and AT-7519 with ABC transporters, which may help elucidate the pharmacokinetic behavior and toxicity of these compounds. Moreover, we show the ability of flavopiridol and SNS-032, but not AT-7519, to overcome ABC transporter-mediated MDR.

Keywords

Cyclin-dependent kinase inhibitor Multidrug resistance ABC transporter AT-7519 Flavopiridol SNS-032 

Abbreviations

ABC

ATP-binding cassette

ABCB1

P-glycoprotein

ABCC1

Multidrug resistance-associated protein 1

ABCG2

Breast cancer resistance protein

CDK

Cyclin-dependent kinase

CDKi

Cyclin-dependent kinase inhibitor

CI

Combination index

DNR

Daunorubicin

DRI

Dose-reduction index

EC50

Median effective antiproliferative concentration

MDCKII

Madin–Darby canine kidney

MDR

Multidrug resistance

MFI

Median fluorescence intensity

MIT

Mitoxantrone

NEM-SG

N-ethylmaleimide-glutathione

PMS

Phenazine methosulfate

RF

Resistance factor

TOP

Topotecan

XTT

XTT sodium salt

Notes

Acknowledgments

This work was supported by the Grant Agency of Charles University in Prague (Grant No. 700912/C/2012 and SVV/2015/260-185). The publication is co-financed by the European Social Fund and the state budget of the Czech Republic (Project No. CZ 1.07/2.2.00/28.0194, the title of the project FAFIS). The manuscript does not contain clinical studies or patient data.

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Daniela Cihalova
    • 1
  • Frantisek Staud
    • 1
  • Martina Ceckova
    • 1
  1. 1.Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec KraloveCharles University in PragueHradec KraloveCzech Republic

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