PMCCPMCCPMCC

Search tips
Search criteria 

Advanced

 
Logo of brjcancerBJC HomepageBJC Advance online publicationBJC Current IssueSubmitting an article to BJCWeb feeds
 
Br J Cancer. 2001 May; 84(10): 1391–1396.
PMCID: PMC2363628

Effect of P-glycoprotein on flavopiridol sensitivity

Abstract

Flavopiridol is the first potent inhibitor of cyclin-dependent kinases (CDKs) to enter clinical trials. Little is known about mechanisms of resistance to this agent. In order to determine whether P-glycoprotein (Pgp) might play a role in flavopiridol resistance, we examined flavopiridol sensitivity in a pair of Chinese hamster ovary cell lines differing with respect to level of Pgp expression. The IC 50 s of flavopiridol in parental AuxB1 (lower Pgp) and colchicine-selected CHRC5 (higher Pgp) cells were 90.2 ± 6.6 nM and 117 ± 2.3 nM, respectively (P< 0.01), suggesting that Pgp might have a modest effect on flavopiridol action. Consistent with this hypothesis, pretreatment with either quinidine or verapamil (inhibitors of Pgp-mediated transport) sensitized CHRC5 cells to the antiproliferative effects of flavopiridol. Because of concern that colony forming assays might not accurately reflect cytotoxicity, we also examined flavopiridol-treated cells by trypan blue staining and flow cytometry. These assays confirmed that flavopiridol was less toxic to cells expressing higher levels of Pgp. Further experiments revealed that flavopiridol inhibited the binding of [3H]-azidopine to Pgp in isolated membrane vesicles, but only at high concentrations. Collectively, these results identify flavopiridol as a weak substrate for Pgp. © 2001 Cancer Research Campaign www.bjcancer.com

Keywords: drug resistance, flavonoids, cyclin-dependent kinase, chemotherapy

Full Text

The Full Text of this article is available as a PDF (191K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Arguello F, Alexander M, Sterry JA, Tudor G, Smith EM, Kalavar NT, Greene JF, Jr, Koss W, Morgan CD, Stinson SF, et al. Flavopiridol induces apoptosis of normal lymphoid cells, causes immunosuppression, and has potent antitumor activity In vivo against human leukemia and lymphoma xenografts. Blood. 1998 Apr 1;91(7):2482–2490. [PubMed]
  • Bech-Hansen NT, Till JE, Ling V. Pleiotropic phenotype of colchicine-resistant CHO cells: cross-resistance and collateral sensitivity. J Cell Physiol. 1976 May;88(1):23–31. [PubMed]
  • Bhalla K, Huang Y, Tang C, Self S, Ray S, Mahoney ME, Ponnathpur V, Tourkina E, Ibrado AM, Bullock G, et al. Characterization of a human myeloid leukemia cell line highly resistant to taxol. Leukemia. 1994 Mar;8(3):465–475. [PubMed]
  • Bible KC, Kaufmann SH. Flavopiridol: a cytotoxic flavone that induces cell death in noncycling A549 human lung carcinoma cells. Cancer Res. 1996 Nov 1;56(21):4856–4861. [PubMed]
  • Bible KC, Kaufmann SH. Cytotoxic synergy between flavopiridol (NSC 649890, L86-8275) and various antineoplastic agents: the importance of sequence of administration. Cancer Res. 1997 Aug 15;57(16):3375–3380. [PubMed]
  • Bible KC, Boerner SA, Kirkland K, Anderl KL, Bartelt D, Jr, Svingen PA, Kottke TJ, Lee YK, Eckdahl S, Stalboerger PG, et al. Characterization of an ovarian carcinoma cell line resistant to cisplatin and flavopiridol. Clin Cancer Res. 2000 Feb;6(2):661–670. [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Bradshaw DM, Arceci RJ. Clinical relevance of transmembrane drug efflux as a mechanism of multidrug resistance. J Clin Oncol. 1998 Nov;16(11):3674–3690. [PubMed]
  • Brüsselbach S, Nettelbeck DM, Sedlacek HH, Müller R. Cell cycle-independent induction of apoptosis by the anti-tumor drug Flavopiridol in endothelial cells. Int J Cancer. 1998 Jul 3;77(1):146–152. [PubMed]
  • Byrd JC, Shinn C, Waselenko JK, Fuchs EJ, Lehman TA, Nguyen PL, Flinn IW, Diehl LF, Sausville E, Grever MR. Flavopiridol induces apoptosis in chronic lymphocytic leukemia cells via activation of caspase-3 without evidence of bcl-2 modulation or dependence on functional p53. Blood. 1998 Nov 15;92(10):3804–3816. [PubMed]
  • Carlson BA, Dubay MM, Sausville EA, Brizuela L, Worland PJ. Flavopiridol induces G1 arrest with inhibition of cyclin-dependent kinase (CDK) 2 and CDK4 in human breast carcinoma cells. Cancer Res. 1996 Jul 1;56(13):2973–2978. [PubMed]
  • Castro AF, Altenberg GA. Inhibition of drug transport by genistein in multidrug-resistant cells expressing P-glycoprotein. Biochem Pharmacol. 1997 Jan 10;53(1):89–93. [PubMed]
  • Cole SP, Downes HF, Slovak ML. Effect of calcium antagonists on the chemosensitivity of two multidrug-resistant human tumour cell lines which do not overexpress P-glycoprotein. Br J Cancer. 1989 Jan;59(1):42–46. [PMC free article] [PubMed]
  • Conseil G, Baubichon-Cortay H, Dayan G, Jault JM, Barron D, Di Pietro A. Flavonoids: a class of modulators with bifunctional interactions at vicinal ATP- and steroid-binding sites on mouse P-glycoprotein. Proc Natl Acad Sci U S A. 1998 Aug 18;95(17):9831–9836. [PubMed]
  • Critchfield JW, Welsh CJ, Phang JM, Yeh GC. Modulation of adriamycin accumulation and efflux by flavonoids in HCT-15 colon cells. Activation of P-glycoprotein as a putative mechanism. Biochem Pharmacol. 1994 Oct 7;48(7):1437–1445. [PubMed]
  • De Azevedo WF, Jr, Mueller-Dieckmann HJ, Schulze-Gahmen U, Worland PJ, Sausville E, Kim SH. Structural basis for specificity and potency of a flavonoid inhibitor of human CDK2, a cell cycle kinase. Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2735–2740. [PubMed]
  • Drees M, Dengler WA, Roth T, Labonte H, Mayo J, Malspeis L, Grever M, Sausville EA, Fiebig HH. Flavopiridol (L86-8275): selective antitumor activity in vitro and activity in vivo for prostate carcinoma cells. Clin Cancer Res. 1997 Feb;3(2):273–279. [PubMed]
  • Ford JM, Yang JM, Hait WN. P-glycoprotein-mediated multidrug resistance: experimental and clinical strategies for its reversal. Cancer Treat Res. 1996;87:3–38. [PubMed]
  • Goldstein LJ, Galski H, Fojo A, Willingham M, Lai SL, Gazdar A, Pirker R, Green A, Crist W, Brodeur GM, et al. Expression of a multidrug resistance gene in human cancers. J Natl Cancer Inst. 1989 Jan 18;81(2):116–124. [PubMed]
  • Greenberger LM, Williams SS, Georges E, Ling V, Horwitz SB. Electrophoretic analysis of P-glycoproteins produced by mouse J774.2 and Chinese hamster ovary multidrug-resistant cells. J Natl Cancer Inst. 1988 Jun 1;80(7):506–510. [PubMed]
  • Greenberger LM, Yang CP, Gindin E, Horwitz SB. Photoaffinity probes for the alpha 1-adrenergic receptor and the calcium channel bind to a common domain in P-glycoprotein. J Biol Chem. 1990 Mar 15;265(8):4394–4401. [PubMed]
  • Hendricks CB, Rowinsky EK, Grochow LB, Donehower RC, Kaufmann SH. Effect of P-glycoprotein expression on the accumulation and cytotoxicity of topotecan (SK&F 104864), a new camptothecin analogue. Cancer Res. 1992 Apr 15;52(8):2268–2278. [PubMed]
  • Kartner N, Evernden-Porelle D, Bradley G, Ling V. Detection of P-glycoprotein in multidrug-resistant cell lines by monoclonal antibodies. Nature. 316(6031):820–823. [PubMed]
  • Kaur G, Stetler-Stevenson M, Sebers S, Worland P, Sedlacek H, Myers C, Czech J, Naik R, Sausville E. Growth inhibition with reversible cell cycle arrest of carcinoma cells by flavone L86-8275. J Natl Cancer Inst. 1992 Nov 18;84(22):1736–1740. [PubMed]
  • Kaye SB. Multidrug resistance: clinical relevance in solid tumours and strategies for circumvention. Curr Opin Oncol. 1998 Aug;10 (Suppl 1):S15–S19. [PubMed]
  • König A, Schwartz GK, Mohammad RM, Al-Katib A, Gabrilove JL. The novel cyclin-dependent kinase inhibitor flavopiridol downregulates Bcl-2 and induces growth arrest and apoptosis in chronic B-cell leukemia lines. Blood. 1997 Dec 1;90(11):4307–4312. [PubMed]
  • Lever JE. Active amino acid transport in plasma membrane vesicles from Simian virus 40-transformed mouse fibroblasts. Characteristics of electrochemical Na+ gradient-stimulated uptake. J Biol Chem. 1977 Mar 25;252(6):1990–1997. [PubMed]
  • Ling V. Multidrug resistance: molecular mechanisms and clinical relevance. Cancer Chemother Pharmacol. 1997;40 (Suppl):S3–S8. [PubMed]
  • Losiewicz MD, Carlson BA, Kaur G, Sausville EA, Worland PJ. Potent inhibition of CDC2 kinase activity by the flavonoid L86-8275. Biochem Biophys Res Commun. 1994 Jun 15;201(2):589–595. [PubMed]
  • Parker BW, Kaur G, Nieves-Neira W, Taimi M, Kohlhagen G, Shimizu T, Losiewicz MD, Pommier Y, Sausville EA, Senderowicz AM. Early induction of apoptosis in hematopoietic cell lines after exposure to flavopiridol. Blood. 1998 Jan 15;91(2):458–465. [PubMed]
  • Patel V, Senderowicz AM, Pinto D, Jr, Igishi T, Raffeld M, Quintanilla-Martinez L, Ensley JF, Sausville EA, Gutkind JS. Flavopiridol, a novel cyclin-dependent kinase inhibitor, suppresses the growth of head and neck squamous cell carcinomas by inducing apoptosis. J Clin Invest. 1998 Nov 1;102(9):1674–1681. [PMC free article] [PubMed]
  • Riordan JR, Deuchars K, Kartner N, Alon N, Trent J, Ling V. Amplification of P-glycoprotein genes in multidrug-resistant mammalian cell lines. Nature. 316(6031):817–819. [PubMed]
  • Safa AR, Glover CJ, Sewell JL, Meyers MB, Biedler JL, Felsted RL. Identification of the multidrug resistance-related membrane glycoprotein as an acceptor for calcium channel blockers. J Biol Chem. 1987 Jun 5;262(16):7884–7888. [PubMed]
  • Schwartz GK, Farsi K, Maslak P, Kelsen DP, Spriggs D. Potentiation of apoptosis by flavopiridol in mitomycin-C-treated gastric and breast cancer cells. Clin Cancer Res. 1997 Sep;3(9):1467–1472. [PubMed]
  • Senderowicz AM, Headlee D, Stinson SF, Lush RM, Kalil N, Villalba L, Hill K, Steinberg SM, Figg WD, Tompkins A, et al. Phase I trial of continuous infusion flavopiridol, a novel cyclin-dependent kinase inhibitor, in patients with refractory neoplasms. J Clin Oncol. 1998 Sep;16(9):2986–2999. [PubMed]
  • Sha EC, Sha MC, Kaufmann SH. Evaluation of 2,6-diamino-N-([1-(1-oxotridecyl)-2-piperidinyl]methyl)- hexanamide (NPC 15437), a protein kinase C inhibitor, as a modulator of P-glycoprotein-mediated resistance in vitro. Invest New Drugs. 1996;13(4):285–294. [PubMed]
  • Van der Bliek AM, Van der Velde-Koerts T, Ling V, Borst P. Overexpression and amplification of five genes in a multidrug-resistant Chinese hamster ovary cell line. Mol Cell Biol. 1986 May;6(5):1671–1678. [PMC free article] [PubMed]
  • Volm M. Multidrug resistance and its reversal. Anticancer Res. 1998 Jul-Aug;18(4C):2905–2917. [PubMed]
  • Waldman T, Zhang Y, Dillehay L, Yu J, Kinzler K, Vogelstein B, Williams J. Cell-cycle arrest versus cell death in cancer therapy. Nat Med. 1997 Sep;3(9):1034–1036. [PubMed]
  • Willingham MC, Cornwell MM, Cardarelli CO, Gottesman MM, Pastan I. Single cell analysis of daunomycin uptake and efflux in multidrug-resistant and -sensitive KB cells: effects of verapamil and other drugs. Cancer Res. 1986 Nov;46(11):5941–5946. [PubMed]
  • Wright J, Blatner GL, Cheson BD. Clinical trials referral resource. Clinical trials of flavopiridol. Oncology (Williston Park) 1998 Jul;12(7):1018–1024. [PubMed]
  • Yang CP, DePinho SG, Greenberger LM, Arceci RJ, Horwitz SB. Progesterone interacts with P-glycoprotein in multidrug-resistant cells and in the endometrium of gravid uterus. J Biol Chem. 1989 Jan 15;264(2):782–788. [PubMed]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK