Intracellular delivery of ceramide lipids via liposomes enhances apoptosis in vitro.
Ceramide lipids have emerged as important intracellular signalling molecules that mediate diverse cellular effects, of which programmed cell death, or apoptosis, has attracted significant interest. Although the exact mechanism(s) by which ceramides trigger apoptosis is not fully understood, there is considerable evidence that they are key mediators of this response. Exogenously applied, cell-permeable ceramides have been shown to induce apoptosis when incubated with cells in culture. We examined here the cytotoxicity of ceramides with varying acyl chain lengths in order to determine whether acyl chain length affects pro-apoptotic activity within the concentration range of 0-100 microM. We found that for C(6)-, C(8)-, C(10)-, C(14)- and C(16)-ceramide, the chain length was inversely proportional to cytotoxic activity, with C(6)-ceramide being most active (IC(50) values in the 3-14 microM range) and C(16)-ceramide being least active (IC(50) values in excess of 100 microM) in the MDA435/LCC6 human breast cancer and J774 mouse macrophage cell lines investigated. Using these two ceramide forms we were able to correlate the observed cytotoxicity with cellular uptake, and we observed that a lack of intracellular delivery may be responsible for the weak activity of C(16)-ceramide. We therefore investigated the possibility of incorporating ceramide lipids into liposome bilayers to enhance this delivery. We demonstrate that stable, ceramide-containing liposomes can be formulated, and that they are cytotoxic when taken up by cells in vitro. These results provide an increased understanding of the differences in cytotoxic activity of exogenous short- and long-chain ceramide lipids, and their incorporation into biologically active liposomal formulations opens new avenues for apoptosis induction.
Shabbits
Jennifer A
JA
Department of Advanced Therapeutics, British Columbia Cancer Research Centre, 601 West 10th Avenue, B.C., V5Z 1L3, Vancouver, Canada.
Mayer
Lawrence D
LD
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000473:pathology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002518:Ceramides / Q000008:administration & dosage* / Q000494:pharmacology; D004337:Drug Carriers; D005260:Female; D006801:Humans; D008081:Liposomes / Q000008:administration & dosage*; D051379:Mice; D013329:Structure-Activity Relationship
D002518:Ceramides; D004337:Drug Carriers; D008081:Liposomes