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Reversal of P-glycoprotein-mediated multidrug resistance of cancer cells by five schizandrins isolated from the Chinese herb Fructus Schizandrae

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Abstract

Purpose

Fructus Schizandrae (FS) is commonly used as a tonic in traditional Chinese medicine. Recently, FS was found to significantly improve liver dysfunction in chronic hepatitis patients. The present study was to assess the reversal effect of five schizandrins and crude extract from FS (named LCC) on multidrug resistance (MDR) of cancer cells, both in vitro and in vivo. Chemically, the five schizandins are derivatives of dibenzo-(a, c)-cyclooctene lignan with distinct structures differing from any known MDR reversal agents.

Methods

A panel of sensitive and resistant cancer cell lines were treated with various concentrations of LCC and schizandrins. Drug sensitivity, accumulation of Doxorubicin (Dox), expression of P-glycoprotein and protein kinase C (PKC), and apoptosis were determined in vitro. The in vivo effect was tested in nude mice grafted with sensitive and resistant human epidermal cancer cell line to vincristine (VCR) (KB, KBv200).

Results

The tested five compounds at 25 μM showed various levels of MDR reversal activity, of which, schizandrin A (Sin A) was the most potent one. Sin A reversed VCR resistance in KBv200 cells, MCF-7/Dox cells and Bel7402 cells by 309-, 38-, and 84-folds, respectively. Also, Sin A reversed the resistance of Dox in the above cancer cell lines. LCC at 25 μg/ml reversed VCR resistance by 619-folds in KBv200, 181-folds in MCF-7/Dox cell line, and 1,563-folds in innate resistance of human hepatic cellular carcinoma Bel7402 cells to VCR. Furthermore, LCC and its active component Sin A potently reversed the cross-resistance to paclitaxel in those cell lines. Both Sin A and LCC markedly increased intracellular Dox accumulation and enhanced apoptosis, down-regulated Pgp protein and mRNA and total PKC expression in MDR cells. Coadministration of LCC (p.o.) significantly potentiated the inhibitory effect of VCR (i.p.) on tumor growth in nude mice bearing KBv200 xenograft.

Conclusions

The LCC and its active component Sin A have remarkable reversal effect on MDR in cancer cells by inhibition of both the function and expression of Pgp and total PKC.

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Abbreviations

FS:

Fructus Schizandrae

Sin A:

Schizandrin A

Sin B:

Schizandrin B

Sin C:

Schizandrin C

Sol A:

Schizandrol A

Sol B:

Schizandrol B

VPL:

Verapamil

Dox:

Doxorubicin

MDR:

Multidrug-resistance

Pgp:

P-glycoprotein

LCC:

Crude extract of Fructus Schizandrae

PKC:

Protein kinase C

VCR:

Vincristine

MTT:

3-(4,5-Dimethylthiazole-2-yle)-2,5-diphenyltetrazolium bromide

Rho 123:

Rhodamine 123

PI:

Propidium iodide

FITC:

Fluorescence isothiocyanate

LPS:

Lipopolysaccharides

CAM:

Camptothecin

ETO:

Etoposide

NBT:

Nitroblue tetrazolium chloride

BCIP:

5-Bromo-4-chloro-3-inDoxyl-phosphate

References

  1. Klopman G, Shi LM, Ramu A (1997) Quantitative structure-activity relationship of multidrug resistance reversal agents. Mol Pharmacol 52:323–334

    PubMed  CAS  Google Scholar 

  2. Gottesman MM, Pastan I (1993) Biochemistry of multidrug resistance mediated by the multidrug transporter. Annu Rev Biochem 62:385–427

    Article  PubMed  CAS  Google Scholar 

  3. Breier A, Barancik M, Sulova Z, Uhrik B (2005) P-glycoprotein–implications of metabolism of neoplastic cells and cancer therapy. Curr Cancer Drug Targets 5:457–468

    Article  PubMed  CAS  Google Scholar 

  4. Gottesman MM (2002) Mechanism of cancer drugs resistance. Annu Rev Med 53:615–627

    Article  PubMed  CAS  Google Scholar 

  5. Takara K, Sakaeda T, Okumura K (2006) An update on overcoming MDR1-mediated multidrug resistance in cancer chemotherapy. Curr Pharm Des 12:273–286

    Article  PubMed  CAS  Google Scholar 

  6. Volm M (1998) Multidrug resistance and its reversal. Anticancer Res 18:2905–2917

    PubMed  CAS  Google Scholar 

  7. Bates SF, Chen C, Robey R, Kang M, Figg WD, Fojo T (2002) Reversal of multidrug resistance: lessons from clinical oncology. Novartis Found Symp 243:83–96; discussion 96–102, 180–5

    Article  PubMed  CAS  Google Scholar 

  8. Ojima I, Geney R, Ungureanu IM, Li D (2002) Medicinal chemistry and chemical biology of new generation taxane antitumor agents. IUBMB Life 53:269–74

    Article  PubMed  CAS  Google Scholar 

  9. Peer D, Margalit R (2006) Fluoxetine and reversal of multidrug resistance. Cancer Lett 237:180-187

    Article  PubMed  CAS  Google Scholar 

  10. O’Brian CA, Ward NE, Stewart JR, Chu F (2001) Prospects for targeting protein kinase C isozymes in the therapy of drug-resistant cancer—an evolving story. Cancer Metastasis Rev 20:95–100

    Article  PubMed  CAS  Google Scholar 

  11. Hardy SP, Valverde MA, Goodfellow HR, Higgins CF, Sepulveda FV (1994) Regulation of volume activated chloride channels by protein kinase C-mediated phosphorylation of P-glycoprotein. Jpn J Physiol 44(Suppl 2):S9–S15

    PubMed  CAS  Google Scholar 

  12. Lelong-Rebel IH, Cardarelli CO, Hardy SP, Valverde MA, Goodfellow HR, Higgins CF, Sepulveda FV (2005) Differential phosphorylation patterns of P-glycoprotein reconstituted into a proteoliposome system: insight into additional unconventional phosphorylation sites. Anticancer Res 25:3925–3935

    PubMed  CAS  Google Scholar 

  13. Masanek U, Stammler G, Volm M (2002) Modulation of multidrug resistance in human ovarian cancer cell lines by inhibition of P-glycoprotein 170 and PKC isoenzymes with antisense oligonucleotides. J Exp Ther Oncol 2:37–41

    Article  PubMed  CAS  Google Scholar 

  14. Beck JF, Brugger D, Brischwein K, Liu C, Bader P, Niethammer D, Gekeler V (2001) Anticancer drug-mediated induction of multidrug resistance-associated genes and protein kinase C isozymes in the T-lymphoblast cell line CCRF-CEM and in blasts from patients with acute lymphoblast leukemias. Jpn J Cancer Res 92:896–903

    PubMed  CAS  Google Scholar 

  15. Brugger D, Brischwein K, Liu C, Bader P, Niethammer D, Gekeler V, Beck JF (2002) Induction of drug resistance and protein kinase C genes in A2780 ovarian cancer cells after incubation with antineoplastic agents at sublethal concentrations. Anticancer Res 22:4229–4232

    PubMed  Google Scholar 

  16. Liu GT (1989) Pharmacological actions and clinical use of Fructus Schizandrae. Chin Med J (Engl) 102:740–749

    CAS  Google Scholar 

  17. Liu GT, Wang GF, Wei HL, Bao TT, Song ZY (1979) A comparison of the protective actions of biphenyl dimethyl-dicarboxylate, transtilbene, alcoholic extracts of Fructus Schizandrae and ganoderma against experimental liver injury in mice. Acta Pharmacol Sin 1:598–604

    Google Scholar 

  18. Liu KT, Cresteil T, Columelli S, Lesca P (1982) Pharmacological properties of dibenzo [a, c] cyclooctene derivatives isolated from Fructus Schizandraee chinensis. II. Induction of phenobarbital-like hepatic monooxygenases. Chem Biol Interact 39:315–330

    Article  PubMed  CAS  Google Scholar 

  19. Lu H, Liu GT (1991) Antioxidant activity of dibenzocyclooctene lignans isolated from Schizandraecea. Planta Med 58:311–315

    Article  Google Scholar 

  20. Liu KT, Lesca P (1982) Pharmacological properties of dibenzo [a, c] cyclooctene derivatives isolated from Fructus Schizandrae chinensis. I. Interaction with rat liver cytochrome P-450 and inhibition of xenobiotics metabolism and mutagenicity. Chem Biol Interact 39:301–314

    Article  PubMed  CAS  Google Scholar 

  21. Huang M, Liu GT (1999) The study of innate drug resistance of human hepatocellular carcinoma Bel7402cll line. Cancer Lett 135:97–105

    Article  PubMed  CAS  Google Scholar 

  22. Chen YY, Shu ZB, Li NL (1976) Studies on Fructus Schisandrae. IV. Isolation and determination of the active components (in lowering high serum ALT levels) of Fructus Schisandrae Sinensis Baill. Sci Sin XIX(1):278–280

  23. Chen YY, Yang YC (1982) Studies on the serum ALT-lowering active component of the Fructus Schisandrae Rubriflora Rhed et Wils. Acta Pharmacol Sin 17:312–315

    Google Scholar 

  24. Fu LW, Zhang YM, Liang YJ, Yang XP, Pan QC (2002) The multidrug resistance of tumour cells was reversed by tetrandrine in vitro and in xenograft derived from human breast adenocarcinoma MCF-7/Dox cells. Eur J Cancer 38:418–426

    Article  PubMed  CAS  Google Scholar 

  25. Zhang XH, Zhang FY, Ji XJ, Li ZY (1994) [Vincristine-resistant human KB cell line and mechanism of multidrug resistance]. Yao Xue Xue Bao 29:246–51

    PubMed  CAS  Google Scholar 

  26. Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB (1987) Evaluation of a tetrazolium-based semi-automated colorimetric assay: assessment of chemosensitivity testing. Cancer Res 47:936–942

    PubMed  CAS  Google Scholar 

  27. Fang M, Zhang HQ, Xue SB, Pang DB, Chi XS (1994) Apoptosis resistance and its reversal in harringtonine resistant cell line. Yao Xue Xue Bao 29:891–898

    PubMed  CAS  Google Scholar 

  28. Telford WG, King LE, Fraker PJ (1994) Rapid quantitation of apoptosis in pure and heterogeneous cell populations using flow cytometry. J Immunol Methods 172:1–16

    Article  PubMed  CAS  Google Scholar 

  29. Ford JM, Bruggemann EP, Pastan I, Gottesman MM, Hait WN (1990) Cellular and biochemical characterization of thioxanthenes for reversal of multidrug resistance in human and murine cell lines. Cancer Res 50:1748–1756

    PubMed  CAS  Google Scholar 

  30. Bonsing BA, Corver WE, Gorsira MC, van Vliet M, Oud PS, Cornelisse CJ, Fleuren GJ (1997) Specificity of seven monoclonal antibodies against p53 evaluated with Western blotting, immunohistochemistry, confocal laser scanning microscopy, and flow cytometry. Cytometry 28:11–24

    Article  PubMed  CAS  Google Scholar 

  31. Lowry OH, Rosenbrough NJ, Farr AL (1951) Protein measurement with the folin phenol reagent. J biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  32. Jin J, Wang FP, Wei H, Liu G (2005) Reversal of multidrug resistance of cancer through inhibition of P-glycoprotein by 5-bromotetrandrine. Cancer Chemother Pharmacol 55:179–188

    Article  PubMed  CAS  Google Scholar 

  33. Laginha KM, Verwoert S, Charrois GJ, Allen TM (2005) Determination of doxorubicin levels in whole tumor and tumor nuclei in murine breast cancer tumors. Clin Cancer Res 11(19pt):6944–6949

    Article  PubMed  CAS  Google Scholar 

  34. Greenlund LJ, Deckwerth TL, Johnson EM Jr (1995) Superoxide dismutase delays neuronal apoptosis: a role for reactive oxygen species in programmed neuronal death. Neuron 14:303–315

    Article  PubMed  CAS  Google Scholar 

  35. Loe DW, Oleschuk CJ, Deeley RG, Cole SP (2000) Structure-activity studies of verapamil analogs that modulate transport of leukotriene C(4) and reduced glutathione by multidrug resistance protein MRP1. Biochem Biophys Res Commun 275:795–803

    Article  PubMed  CAS  Google Scholar 

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Acknowledgment

This work was supported by the Natural Science Foundation of China (No. 30630069).

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Author notes

  1. Min Huang

    Present address: Division of Oncology, Department of Internal Medicine, Stanford University, Stanford, CA, 94301, USA

  2. Jing Jin

    Present address: Welsh School of Pharmacy, Cardiff University, Cardiff, UK

Authors and Affiliations

  1. Department of Pharmacology, Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, 1 Xian Nong Tan Street, 100050, Beijing, China

    Hua Sun & Geng Tao Liu

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  1. Min Huang
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  2. Jing Jin
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  3. Hua Sun
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  4. Geng Tao Liu
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Corresponding author

Correspondence to Geng Tao Liu.

Additional information

Min Huang and Jing Jin have contributed equally to this work.

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Huang, M., Jin, J., Sun, H. et al. Reversal of P-glycoprotein-mediated multidrug resistance of cancer cells by five schizandrins isolated from the Chinese herb Fructus Schizandrae . Cancer Chemother Pharmacol 62, 1015–1026 (2008). https://doi.org/10.1007/s00280-008-0691-0

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