Apoptosis, necrosis and cell proliferation-inhibition by cyclosporine A in U937 cells (a human monocytic cell line)

Pharmacol Res. 2006 Mar;53(3):293-302. doi: 10.1016/j.phrs.2005.12.007. Epub 2006 Feb 2.

Abstract

The immunosuppressive drug cyclosporine A (CsA) has been used in both organ transplantation and the treatment of autoimmune disorders. However, the drug causes adverse effects in the kidney, liver and nervous system, characterized by cellular loss in the affected area. Apoptosis has been shown to play a role in CsA-induced cytotoxicity. Because permeabilization of the mitochondrial membrane is a common criterion in most apoptotic settings in vertebrate cells, here we evaluated whether CsA causes loss of mitochondrial function in the pathway leading to cellular cytotoxicity. We found that CsA caused a concentration- and time-dependent loss of cell viability in the U937 cell line. Treatment of cells at a dose of 10 microM CsA resulted in G0/G1 arrest with a concurrent decrease in the number of cells in the S and G2/M phases of the cell cycle. In mechanistic studies related to the loss of viability, treating cells with 10 microM CsA for 24 h resulted in both DNA fragmentation and an increase of annexin-V-positive cells. CsA treatment also increased activity of the cysteine protease caspase-3, decreased the mitochondrial membrane potential and induced the release of cytochrome c into the cytosol. Furthermore, CsA treatment increased the number of cells in the sub-G0/G1 peak, indicative of a reduction in DNA, although this increase was not observed when cells were pre-treated with a broad caspase inhibitor. In the study, we also found that a higher dose of CsA induces LDH release when the cells were incubated for a longer period. Taken together, these data suggest that the mode of cell death induced by CsA is dose- and time-dependent. Short-term incubation with lower doses of CsA arrests cell growth; this arrest overlaps with the occurrence of apoptosis and then with necrosis after longer treatment periods with higher doses of CsA.

MeSH terms

  • Apoptosis / drug effects*
  • Aspartic Acid / analogs & derivatives
  • Aspartic Acid / pharmacology
  • Caspase 3
  • Caspase Inhibitors
  • Caspases / metabolism
  • Cell Proliferation / drug effects*
  • Cell Survival / drug effects
  • Cyclosporine / toxicity*
  • Cytochromes c / metabolism
  • Dose-Response Relationship, Drug
  • Humans
  • Immunosuppressive Agents / toxicity*
  • Membrane Potentials
  • Mitochondria / drug effects*
  • Mitochondria / enzymology
  • Necrosis
  • Protease Inhibitors / pharmacology
  • Time Factors
  • U937 Cells

Substances

  • Caspase Inhibitors
  • Immunosuppressive Agents
  • Protease Inhibitors
  • benzyloxycarbonyl-Asp-CH2OC(O)-2,6-dichlorobenzene
  • Aspartic Acid
  • Cyclosporine
  • Cytochromes c
  • CASP3 protein, human
  • Caspase 3
  • Caspases