Ceramide-containing liposomes with doxorubicin: time and cell-dependent effect of C6 and C12 ceramide

// Anders Overbye 1, 2 , Ann Mari Holsaeter 3 , Markus Fusser 4 , Natasa Skalko-Basnet 3 , Tore-Geir Iversen 1, 2 , Maria Lyngaas Torgersen 1, 2 , Tonje Sonstevold 1, 2 , Olav Engebraaten 4, 5 , Kjersti Flatmark 4, 5 , Gunhild Mari Maelandsmo 3, 4 , Tore Skotland 1, 2 and Kirsten Sandvig 1, 2, 6 1 Centre for Cancer Biomedicine, Faculty Division Norwegian Radium Hospital, University of Oslo, Oslo, Norway 2 Department of Molecular Cell Biology, Institute for Cancer Research, The Norwegian Radium Hospital, Oslo University Hospital, Oslo, Norway 3 Drug Transport and Delivery Research Group, Department of Pharmacy, Faculty of Health Sciences, University of Tromso – The Arctic University of Norway, Tromso, Norway 4 Department of Tumour Biology, Institute for Cancer Research, The Norwegian Radium Hospital, Oslo University Hospital, Oslo, Norway 5 Institute for Clinical Medicine, Faculty of Medicine, University of Oslo, Oslo, Norway 6 Department of Biosciences, University of Oslo, Oslo, Norway Correspondence to: Anders Overbye, email: anders.overbye@rr-research.no Keywords: liposomes, ceramide, doxorubicin, mice tumors, cell studies Received: March 24, 2017 Accepted: June 17, 2017 Published: August 12, 2017 ABSTRACT Doxorubicin, a widely used chemotherapeutic drug, has several potential high-risk side effects including cardiomyopathy. Furthermore, cellular resistance to this drug develops with time. By using liposomes as carrier vesicles both the side effects and drug resistance might be avoided. In this study we have investigated the cytotoxic effect of doxorubicin encapsulated in liposomes with and without ceramides containing 6 or 12 carbon atoms in the N-amidated fatty acyl chains. The short-chain ceramide species were included in the liposomal compositions due to their pro-apoptotic properties, which might cause a synergistic anticancer effect. We demonstrate that the ceramide species enhance the liposomal doxorubicin toxicity in a cell-specific manner. The C6-ceramide effect is most pronounced in cervical cancer cells (HeLa) and colon cancer cells (HCT116), whereas the C12-ceramide effect is strongest in breast cancer cells (MDA-MB-231). Moreover, the study reveals the importance of investigating cell toxicity at several time points and in different cell-lines, to assess drug-and formulation-induced cytotoxic effects in vitro . Furthermore, our data show that the cytotoxicity obtained with the nanocarriers in vitro , does not necessarily reflect their ability to inhibit tumor growth in vivo . We speculate that the larger effect of Caelyx® than our liposomes in vivo is due to a greater in vivo stability of Caelyx®.