Characterization of the ATP-dependent LTC4 transporter in cisplatin-resistant human KB cells

Biochem Biophys Res Commun. 1996 Sep 4;226(1):158-65. doi: 10.1006/bbrc.1996.1326.

Abstract

An active efflux pump for cis-diamminedichloroplatinum(II) (cisplatin) has been identified in cisplatin-resistant KCP-4 cells isolated from human epidermoid carcinoma KB-3-1 cells. The adenosine triphosphate(ATP)-dependent transport of leukotriene C4 (LTC4), an endogenous substrate for the glutathione S-conjugate export pump(GS-X pump), has been found in membrane vesicles prepared from KCP-4 cells. Multidrug resistance-associated protein (MRP) has also been identified as an ATP-dependent LTC4 transporter. To examine whether the GS-X pump expressed in KCP-4 cells in MRP, we investigated the expression of MRP in KCP-4 cells and compared the LTC4 transporting activity of GS-X pump expressed in KCP-4 cells with that of MRP. The level of MRP gene expression in KCP-4 cells was low and similar to that in KB-3-1 cells. MRP was not detected in membrane vesicles prepared from KB-3-1 and KCP-4 cells by immunoblot analysis. The ATP-dependent transport of LTC4 in KCP-4 and C-A120 vesicles showed saturable kinetics with an apparent Km of 0.18 microM and 0.25 microM, respectively. [3H]LTC4 transport in KCP-4 vesicles was more inhibited by 2,4-dinitrophenyl-S-glutathione(DNP-SG), bis-(glutathionato)-platinum(II) (GS-platinum) complex and glutathione disulfide(GS-SG) and less by LTD4 compared with that in C-A120 vesicles. The character of the LTC4 transporter expressed in KCP-4 vesicles is similar but not identical to that of MRP. Our results suggest that a glutathione S-conjugate export pump which is different from MRP exists in cisplatin-resistant KCP-4 cells.

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily B, Member 1 / genetics
  • ATP-Binding Cassette Transporters / genetics
  • Adenosine Triphosphate / metabolism*
  • Amino Acid Sequence
  • Biological Transport
  • Cell Membrane / metabolism
  • Cisplatin / pharmacology*
  • Drug Resistance, Neoplasm
  • Glutathione / metabolism
  • Humans
  • Leukotriene C4 / metabolism*
  • Molecular Sequence Data
  • Multidrug Resistance-Associated Proteins
  • Osmolar Concentration
  • RNA, Messenger / genetics
  • Tritium
  • Tumor Cells, Cultured
  • Verapamil / metabolism

Substances

  • ATP Binding Cassette Transporter, Subfamily B, Member 1
  • ATP-Binding Cassette Transporters
  • Multidrug Resistance-Associated Proteins
  • RNA, Messenger
  • Tritium
  • Leukotriene C4
  • Adenosine Triphosphate
  • Verapamil
  • Glutathione
  • Cisplatin