C-Phycocyanin, a selective cyclooxygenase-2 inhibitor, induces apoptosis in lipopolysaccharide-stimulated RAW 264.7 macrophages

Biochem Biophys Res Commun. 2003 May 2;304(2):385-92. doi: 10.1016/s0006-291x(03)00586-2.

Abstract

C-Phycocyanin (C-PC) is one of the major biliproteins of Spirulina platensis, a blue green algae, with antioxidant and radical scavenging properties. It is also known to exhibit anti-inflammatory and anti-cancer properties. However, the mechanism of action of C-PC is not clearly understood. Previously, we have shown that C-PC selectively inhibits cyclooxygenase-2 (COX-2), an inducible isoform that is upregulated during inflammation and cancer. In view of the reported induction of apoptosis in cancer cells by cyclooxygenase-2 inhibitors, the present study is undertaken to test the effect of C-PC on LPS stimulated RAW 264.7 mouse macrophage cell line. These studies have shown a dose dependent reduction in the growth and multiplication of macrophage cell line by C-PC. This decrease in cell number appears to be mediated by C-PC induced apoptosis as evidenced by flow cytometric and confocal microscopic studies. Cells treated with 20 micro M C-PC showed typical nuclear condensation and 16.6% of cells in sub-G(o)/G(1) phase. These cells also showed DNA fragmentation in a dose dependent manner. The studies on poly(ADP ribose) polymerase (PARP) cleavage showed typical fragmentation pattern in C-PC treated cells. This C-PC induced apoptosis in RAW 264.7 cells appears to be mediated by the release of cytochrome c from mitochondria and independent of Bcl-2 expression. These effects of C-PC on RAW 264.7 cells may be due to reduced PGE(2) levels as a result of COX-2 inhibition.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis*
  • Cell Division / drug effects
  • Cell Line
  • Cell Nucleus / ultrastructure
  • Cell Survival / drug effects
  • Cyclooxygenase 1
  • Cyclooxygenase 2
  • Cyclooxygenase 2 Inhibitors
  • Cyclooxygenase Inhibitors / pharmacology*
  • Cytochrome c Group / metabolism
  • DNA Fragmentation
  • Dinoprostone / biosynthesis
  • Flow Cytometry
  • Isoenzymes / antagonists & inhibitors*
  • Isoenzymes / metabolism
  • Lipopolysaccharides / pharmacology
  • Macrophages / cytology
  • Macrophages / drug effects*
  • Macrophages / enzymology
  • Macrophages / metabolism
  • Membrane Proteins
  • Mice
  • Microscopy, Confocal
  • Mitochondria / metabolism
  • Phycocyanin / pharmacology*
  • Poly(ADP-ribose) Polymerases / metabolism
  • Prostaglandin-Endoperoxide Synthases / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / metabolism

Substances

  • Cyclooxygenase 2 Inhibitors
  • Cyclooxygenase Inhibitors
  • Cytochrome c Group
  • Isoenzymes
  • Lipopolysaccharides
  • Membrane Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • Phycocyanin
  • Cyclooxygenase 1
  • Cyclooxygenase 2
  • Prostaglandin-Endoperoxide Synthases
  • Ptgs1 protein, mouse
  • Poly(ADP-ribose) Polymerases
  • Dinoprostone