Cancer Chemotherapy and Pharmacology

, Volume 35, Issue 3, pp 267–269 | Cite as

The structure of P-glycoprotein and the secretion of lysosomal enzymes in multidrug-resistant cells

  • Leonard Warren
  • Anna Malarska
  • Jean-Claude Jardillier
Short Communication Multidrug Resistance, Lysosomal Enzymes, Mutant P-Glycoprotein

Abstract

We have previously demonstrated that multidrug-resistant cells have a lower content of lysosomal enzymes, a consequence of an increased rate of secretion. The question was therefore to know whether an intact functional P-glycoprotein was necessary for expression of this property. Control NIH3T3 andmdr1-gene-transfected cells (pHaMDR1) were used together with 2 variants either lacking 23 amino acids at the carboxyl terminal (pHaMDRC 23) or in which 4 extra amino acids are inserted (pHaMDRBL2). Transfected and variant cells exhibited reduced uptake of [3H]-vinblastine and [3H]-daunomycin, a finding consistent with their drug resistance. By contrast, only pHaMDR1 cells had a reduced level ofN-acetyl glucosaminidase that paralleled an increased rate of secretion of the same enzyme. The mutant cells secreted lysosomal enzyme at the same rate and had the same intracellular lysosomal enzyme content as NIH3T3 cells. Abnormal behavior of lysosomal enzymes in multidrug-resistant cells therefore seemed to require an intact P-glycoprotein molecule. Although sequestration in lysosomes and then secretion of drugs may possibly contribute to protection, it would not be an essential component of multidrug resistance.

Key words

Multidrug resistance Lysosomal enzymes Mutant P-glycoprotein 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Ambukar SV, Lelong IH, Zhang J, Cardarelli CO, Gottesman MM, Pastan I (1992) Partial purification and reconstitution of the human multidrug-resistance pump: characterization of the drug-stimulatable ATP hydrolysis. Proc Natl Acad Sci USA 89: 8472Google Scholar
  2. 2.
    Bradley G, Juranka PE, Ling V (1988) Mechanism of multidrug resistance. Biochim Biophys Acta 948: 87Google Scholar
  3. 3.
    Currier SJ, Ueda K, Willingham MC, Pastan I, Gottesman MM (1989) Deletion and insertion mutants of the multidrug transporter. J Biol Chem 264: 376Google Scholar
  4. 4.
    Gottesman MM (1993) How cancer cells evade chemotherapy: sixteenth Richard and Hinda Rosenthal Foundation award lecture. Cancer Res 53: 74Google Scholar
  5. 5.
    Gottesman MM, Pastan I (1993) Biochemistry of multidrug resistance mediated by the multidrug transporter. Annu Rev Biochem 62: 385Google Scholar
  6. 6.
    Hammond JR, Johnstone RM, Gros P (1989) Enhanced efflux of [3H]vinblastine from chinese hamster ovary cells transfected with a full-length complementary DNA clone for themdr1 gene. Cancer Res 49: 3867Google Scholar
  7. 7.
    Kane SE, Troen BR, Gal S, Ueda K, Pastan I, Gottesman MM (1988) Use of a cloned multidrug resistance gene for coamplification and overproduction of major excreted protein, a transformation-regulated secreted acid protease. Mol Cell Biol 8: 3316Google Scholar
  8. 8.
    Klohs WD, Steinkampf RW (1988) The effect of lysomotropic agents and secretory inhibitors on anthracycline retention and activity in multiple drug-resistant cells. Mol Pharmacol 34: 180Google Scholar
  9. 9.
    Marquardt D, Center MS (1991) Involvement of vacuolar H+-adenosine triphosphatase activity in multidrug resistance in HL60 cells. J Natl Cancer Inst 83: 1098Google Scholar
  10. 10.
    Sehested M, Skovsgaard T, Van Deurs B, Winther-Nielsen H (1987) Increased plasma membrane traffic in daunorubicin resistant P388 leukaemic cells. Effect of daunorubicin and verapamil. Br J Cancer 56: 747Google Scholar
  11. 11.
    Sehested M, Skovsgaard T, Roed H (1988) The carboxylic ionophore monensin inhibits active drug efflux and modulates in vitro resistance in daunorubicin resistant Ehrlich ascites tumor cells. Biochem Pharmacol 37: 3305Google Scholar
  12. 12.
    Slapak CA, Lecerf JM, Daniel JC, Levy SB (1992) Energy-dependent accumulation of daunorubicin into subcellular compartments of human leukemia cells and cytoplasts. J Biol Chem 267: 10638Google Scholar
  13. 13.
    Stow MW, Warr JR (1993) Reduced influx is a factor in accounting for reduced vincristine accumulation in certain verapamil-hypersensitive multidrug-resistant CHO cell lines. FEBS Lett 320: 87Google Scholar
  14. 14.
    Ueda K, Cardarelli C, Gottesman MM, Pastan I (1987) Expression of a full-length cDNA for the humanmdr1 gene confers resistance to colchicine, doxorubicin, and vinblastine. Proc Natl Acad Sci USA 84: 3004Google Scholar
  15. 15.
    Van der Blick AM, Borst P (1989) Multidrug resistance. Adv Cancer Res 52: 165Google Scholar
  16. 16.
    Vendrik CPJ, Bergers JJ, Dejong WH, Steerenberg PA (1992) Resistance to cytostatic drugs at the cellular level. Cancer Chemother Pharmacol 29: 413Google Scholar
  17. 17.
    Warren L (1989) Stimulated secretion of lysosomal enzymes by cells in culture. J Biol Chem 264: 8835Google Scholar
  18. 18.
    Warren L, Jardillier J-C, Ordentlich P (1991) Secretion of lysosomal enzymes by drug-sensitive and multiple drug-resistant cells. Cancer Res 51: 1996Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Leonard Warren
    • 1
  • Anna Malarska
    • 1
  • Jean-Claude Jardillier
    • 2
  1. 1.The Wistar Institute of Anatomy and BiologyPhiladelphiaUSA
  2. 2.GIBSA, EA 1238, Institut Jean Godinot and UFR de PharmacieUniversity of ReimsReims CedexFrance

Personalised recommendations