Improving Intracellular Doxorubicin Delivery Through Nanoliposomes Equipped with Selective Tumor Cell Membrane Permeabilizing Short-Chain Sphingolipids
To improve nanoliposomal-doxorubicin (DoxNL) delivery in tumor cells using liposome membrane-incorporated short-chain sphingolipids (SCS) with selective membrane-permeabilizing properties. DoxNL bilayers contained synthetic short-chain derivatives of known membrane microdomain-forming sphingolipids; C8-glucosylceramide (C8-GluCer), C8-galactosylceramide (C8-GalCer) or C8-lactosylceramide (C8-LacCer).
DoxNL enriched with C8-GluCer or C8-GalCer were developed, optimized and characterized with regard to size, stability and drug retention. In vitro cytotoxic activity was studied in a panel of human tumor cell lines and normal cells. Intracellular Dox delivery was measured by flow cytometry and visualized by fluorescence microscopy. For a further understanding of the involved drug delivery mechanism confocal microscopy studies addressed the cellular fate of the nanoliposomes, the SCS and Dox in living cells.
C8-LacCer-DoxNL aggregated upon Dox loading. In tumor cell lines SCS-DoxNL with C8-GluCer or C8-GalCer demonstrated strongly increased Dox delivery and cytotoxicity compared to standard DoxNL. Surprisingly, this effect was much less pronounced in normal cells. Nanoliposomes were not internalized, SCS however transfered from the nanoliposomal bilayer to the cell membrane and preceded cellular uptake and subsequent nuclear localization of Dox.
C8-GluCer or C8-GalCer incorporated in DoxNL selectively improved intracellular drug delivery upon transfer to tumor cell membranes by local enhancement of cell membrane permeability.
KEY WORDSdoxorubicin-nanoliposome short chain sphingolipid Tumor cell membrane permeabilization
- D:PL ratio
Drug to phospholipid initial mass ratio
Fetal calf serum
Hydrogenated soy phosphatidylcholine
Mean of fluorescence intensity
- NBD PE
Short chain sphingolipids
Standard error of the mean
Tumor cell membrane modulation
- 5.Mayer LD, Bally MB, Cullis PR, Wilson SL, Emerman JT. Comparison of free and liposome encapsulated Dox tumor drug uptake and antitumor efficacy in the SC115 murine mammary tumor. Cancer Lett. 1999;53(2–3):183–90.Google Scholar
- 16.O’Brien ME, Wigler N, Inbar M, Rosso R, Grischke E, Santoro A, et al. CAELYX Breast Cancer Study Group, Reduced cardiotoxicity and comparable efficacy in a phase III trial of pegylated liposomal Dox HCl (CAELYX/DoxilÂ®) versus conventional Dox for first-line treatment of metastatic breast cancer. Ann Oncol. 2004;15(3):440–9.PubMedCrossRefGoogle Scholar
- 18.Northfelt DW, Dezube BJ, Thommes JA, Miller BJ, Fischl MA, Friedman-Kien A, et al. Pegylated-liposomal Dox versus Dox, bleomycin, and vincristine in the treatment of AIDS-related Kaposi’s sarcoma: results of a randomized phase III clinical trial. J Clin Oncol. 1998;16(7):2445–51.PubMedGoogle Scholar
- 20.Hussein MA, Anderson KC. Role of liposomal anthracyclines in the treatment of multiple myeloma. Semin Oncol. 2006;31(6 Suppl 13):147–60.Google Scholar
- 21.Alberts DS, Liu PY, Wilczynski SP, Clouser MC, Lopez AM, Michelin DP, et al. Randomized trial of pegylated liposomal Dox (PLD) plus carboplatin versus carboplatin in platinum-sensitive (PS) patients with recurrent epithelial ovarian or peritoneal carcinoma after failure of initial platinum-based chemotherapy (Southwest Oncology Group Protocol S0200). Gynecol Oncol. 2008;108(1):90–4.PubMedCrossRefGoogle Scholar
- 24.Judson I, Radford JA, Harris M, Blay JY, van Hoesel Q, le Cesne A, et al. Randomised phase II trial of pegylated liposomal doxorubicin (DOXIL/CAELYX) versus doxorubicin in the treatment of advanced or metastatic soft tissue sarcoma: a study by the EORTC Soft Tissue and Bone Sarcoma Group. Eur J Cancer. 2001;37(7):870–7.PubMedCrossRefGoogle Scholar
- 37.Koning GA, Morselt HW, Velinova MJ, Gorter A, Allen TM, Zalipsky S, Et al.,Selective transfer of a lipophilic prodrug of 5-fluorodeoxyuridine from immunoliposomes to colon cancer cells. Biochim Biophys Acta. 1999;1420(1–2):153–67.Google Scholar