alpha(v)beta(3) Integrin-mediated drug resistance in human laryngeal carcinoma cells is caused by glutathione-dependent elimination of drug-induced reactive oxidative species

Mol Pharmacol. 2008 Jul;74(1):298-306. doi: 10.1124/mol.107.043836. Epub 2008 Apr 25.

Abstract

As a model for determination of the role of integrins in drug resistance, we used alpha(v)beta(3) integrin-negative human laryngeal carcinoma cell line (HEp2) and three HEp2-derived cell clones with a gradual increase of alpha(v)beta(3) integrin expression. The alpha(v)beta(3) integrin expression protects cells from cisplatin, mitomycin C, and doxorubicin. In HEp2-alpha(v)beta(3) integrin-expressing cells, the constitutive expression of Bcl-2 protein and the level of glutathione (GSH) were increased compared with HEp2 cells. Pretreatment of HEp2-alpha(v)beta(3) integrin-expressing cells with an inhibitor of GSH synthesis, buthionine sulfoximine (BSO), decreased the level of GSH and partially reverted drug resistance to all above-mentioned drugs, but it did not influence the expression of Bcl-2. Sensitivity to selected anticancer drugs did not change with overexpression of Bcl-2 in HEp2 cells, nor with silencing of Bcl-2 in HEp2-alpha(v)beta(3) integrin-expressing cells, indicating that Bcl-2 is not involved in resistance mechanism. There was no difference in DNA platination between HEp2 and HEp2-alpha(v)beta(3) integrin-expressing cells, indicating that the mechanism of drug resistance is independent of cisplatin detoxification by GSH. A strong increase of reactive oxidative species (ROS) formation during cisplatin or doxorubicin treatment in HEp2 cells was reduced in HEp2-alpha(v)beta(3) integrin-expressing cells. Since this increased elimination of ROS could be reverted by GSH depletion, we concluded that multidrug resistance is the consequence of GSH-dependent increased ability of alpha(v)beta(3)-expressing cells to eliminate drug-induced ROS.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibiotics, Antineoplastic / pharmacology
  • Antineoplastic Agents / pharmacology
  • Buthionine Sulfoximine / pharmacology
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cisplatin / pharmacology
  • Clone Cells
  • Doxorubicin / pharmacology
  • Drug Resistance, Neoplasm*
  • Enzyme Inhibitors / pharmacology
  • Formazans / metabolism
  • Glutathione / analysis
  • Glutathione / metabolism*
  • Humans
  • Integrin alphaVbeta3 / metabolism*
  • Laryngeal Neoplasms / pathology
  • Mitomycin / pharmacology
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Reactive Oxygen Species / metabolism*
  • Tetrazolium Salts / metabolism

Substances

  • Antibiotics, Antineoplastic
  • Antineoplastic Agents
  • Enzyme Inhibitors
  • Formazans
  • Integrin alphaVbeta3
  • Proto-Oncogene Proteins c-bcl-2
  • Reactive Oxygen Species
  • Tetrazolium Salts
  • MTT formazan
  • Buthionine Sulfoximine
  • Mitomycin
  • Doxorubicin
  • Glutathione
  • Cisplatin