Ceramide methylaminoethylphosphonate (
CMAEPn) was isolated from eastern oyster ( Crassostrea virginica ) and screened against in vitro and in vivo angiogenesis and against MCF-7 and MDA-MB-435s
breast cancer cell lines. In vitro angiogenesis was evaluated by the
vascular endothelial growth factor (
VEGF)-induced human umbilical vein endothelial cell (HUVEC) tube formation assay. MCF-7 and MDA-MB-435s cell viability was evaluated by the CellTiter 96 AQ(ueous) One
Solution Cell Proliferation assay. Apoptosis was evaluated by the
caspase-9 assay, autophagy by
acridine orange staining and
beclin-1 level. Our study indicates that
CMAEPn at 50 microM inhibited
VEGF-induced tube formation by HUVEC. The viability of MCF-7 and MDA-MB-435s
breast cancer cells exposed to 125 microM
CMAEPn for 48 h was reduced to 76 and 85%, respectively. The viability of MCF-7 and MDA-MB-435s cells exposed to 250 microM
CMAEPn for 48 h under the same conditions was reduced to 38 and 45%, respectively.
CMAEPn at 125 microM inhibited
VEGF-induced MDA-MB-435s cell migration and invasion.
CMAEPn at 125 microM also decreased
VEGF,
EGF levels in the
conditioned media, PI3K, IkappaB phosphorylation and degradation in the cytoplasmic extracts, and NFkappaB nuclear translocation. Both
acridine orange staining and
beclin-1 indicated autophagic cell death in MCF-7 and MDA-MB-435s cells, respectively. In vivo,
CMAEPn at 30 mg/kg
body weight inhibited bFGF-induced angiogenesis and caused a 57% reduction in
hemoglobin levels in the
matrigel plug assay within 7 days. This is the first report on
CMAEPn-inhibited angiogenesis both in vitro and in vivo.