Multidrug resistance protein (MRP) 1 and MRP3 attenuate cytotoxic and transactivating effects of the cyclopentenone prostaglandin, 15-deoxy-Delta(12,14)prostaglandin J2 in MCF7 breast cancer cells

Biochemistry. 2003 May 13;42(18):5429-37. doi: 10.1021/bi027347u.

Abstract

One of the most potent cyclopentenone prostaglandins, 15-deoxy-Delta(12,14)prostaglandin J(2) (15-d-PGJ(2)), has been shown to be cytotoxic in some tumor cells and, as a ligand of peroxisome proliferator activated receptor gamma (PPARgamma), to influence the transcriptional regulation of several genes. We examined whether a glutathione conjugate of 15-d-PGJ(2), 15-d-PGJ(2)-SG, is formed and if the glutathione conjugate efflux pumps, MRP1 and MRP3, could transport this conjugate, thereby attenuating the cytotoxicity and transactivating activity of 15-d-PGJ(2) in MCF7 breast cancer cells. Formation of 15-d-PGJ(2)-SG was demonstrated both in vitro and in cells, and its structure was determined by ESI/MS and NMR. Expression of MRP1 and MRP3 was achieved by stable transduction of parental MCF7 cells. Membrane vesicles derived from these cells supported efficient, ATP-dependent transport of 15-d-PGJ(2)-SG (K(M) 1.4 and 2.9 microM for MRP1 and MRP3, respectively). When compared with parental, MRP-minus MCF7 cells, expression of MRP1 and MRP3 conferred approximately 2-fold protection from 15-d-PGJ(2) cytotoxicity. 15-d-PGJ(2)-mediated transcriptional activation was evaluated in cells transiently transfected with a reporter gene under the transcriptional control of a PPAR responsive element. Treatment of parental MCF7 cells with 15-d-PGJ(2) resulted in a time-dependent induction of reporter gene activity-induction that was measurable with concentrations of added 15-d-PGJ(2) as low as 100 nM. In contrast, expression of MRP1 or MRP3 abolished 15-d-PGJ(2)-dependent reporter gene induction. Depletion of intracellular glutathione reversed MRP1- and MRP3-mediated attenuation of 15-d-PGJ(2) cytotoxicity and transactivation. These data indicate that MRP1 and MRP3 can modulate the biological effects of 15-d-PGJ(2), and likely other cyclopentenone prostaglandins, in a glutathione-dependent manner. The results are consistent with a mechanism for the attenuation of the biological activities of 15-d-PGJ(2) that involves the formation and active efflux of its glutathione conjugate, 15-d-PGJ(2)-SG.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B, Member 1 / physiology*
  • Adenosine Triphosphate / metabolism
  • Biological Transport
  • Breast Neoplasms / metabolism*
  • Breast Neoplasms / pathology
  • Cell Membrane / chemistry
  • Cell Membrane / metabolism
  • Cell Survival
  • Drug Resistance, Neoplasm
  • Gene Expression Regulation, Neoplastic / drug effects
  • Glutathione / analogs & derivatives
  • Glutathione / metabolism
  • Humans
  • Immunologic Factors / metabolism*
  • Immunologic Factors / pharmacology
  • Inhibitory Concentration 50
  • Kinetics
  • Luciferases / metabolism
  • Magnetic Resonance Spectroscopy
  • Methotrexate / pharmacology
  • Multidrug Resistance-Associated Proteins / physiology*
  • Prostaglandin D2 / analogs & derivatives
  • Prostaglandin D2 / metabolism*
  • Prostaglandin D2 / pharmacology
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Spectrometry, Mass, Electrospray Ionization
  • Transcription Factors / metabolism
  • Transcriptional Activation*
  • Transfection
  • Tumor Cells, Cultured

Substances

  • 15-deoxy-delta(12,14)-prostaglandin J2
  • ATP Binding Cassette Transporter, Subfamily B, Member 1
  • Immunologic Factors
  • Multidrug Resistance-Associated Proteins
  • Receptors, Cytoplasmic and Nuclear
  • Transcription Factors
  • multidrug resistance-associated protein 3
  • Adenosine Triphosphate
  • Luciferases
  • Glutathione
  • Prostaglandin D2
  • Methotrexate