title: Cationic PEGylated liposomes incorporating an antimicrobial peptide tilapia hepcidin 2-3: an adjuvant of epirubicin to overcome multidrug resistance in cervical cancer cells.
abstract: Antimicrobial peptides (AMPs) have been recently evaluated as a new generation of adjuvants in cancer chemotherapy. In this study, we designed PEGylated liposomes encapsulating epirubicin as an antineoplastic agent and tilapia hepcidin 2-3, an AMP, as a multidrug resistance (MDR) transporter suppressor and an apoptosis/autophagy modulator in human cervical cancer HeLa cells. Cotreatment of HeLa cells with PEGylated liposomal formulation of epirubicin and hepcidin 2-3 significantly increased the cytotoxicity of epirubicin. The liposomal formulations of epirubicin and/or hepcidin 2-3 were found to noticeably escalate the intracellular H2O2and O2-levels of cancer cells. Furthermore, these treatments considerably reduced the mRNA expressions of MDR protein 1, MDR-associated protein (MRP) 1, and MRP2. The addition of hepcidin 2-3 in liposomes was shown to markedly enhance the intracellular epirubicin uptake and mainly localized into the nucleus. Moreover, this formulation was also found to trigger apoptosis and autophagy in HeLa cells, as validated by significant increases in the expressions of cleaved poly ADP ribose polymerase, caspase-3, caspase-9, and light chain 3 (LC3)-II, as well as a decrease in mitochondrial membrane potential. The apoptosis induction was also confirmed by the rise in sub-G1 phase of cell cycle assay and apoptosis percentage of annexin V/propidium iodide assay. We found that liposomal epirubicin and hepcidin 2-3 augmented the accumulation of GFP-LC3 puncta as amplified by chloroquine, implying the involvement of autophagy. Interestingly, the partial inhibition of necroptosis and the epithelial-mesenchymal transition by this combination was also verified. Altogether, our results provide evidence that coincubation with PEGylated liposomes of hepcidin 2-3 and epirubicin caused programmed cell death in cervical cancer cells through modulation of multiple signaling pathways, including MDR transporters, apoptosis, autophagy, and/or necroptosis. Thus, this formulation may provide a new platform for the combined treatment of traditional chemotherapy and hepcidin 2-3 as a new adjuvant for effective MDR reversal.
authors.affiliation: Department and Institute of Pharmacology, National Yang-Ming University.
authors.forename: Vivian
authors.initials: V
authors.lastname: Juang
authors.affiliation: Department of Biological Sciences and Technology, National University of Tainan.
authors.forename: Hsin-Pin
authors.initials: HP
authors.lastname: Lee
authors.affiliation: Department and Institute of Pharmacology, National Yang-Ming University; Department of Medical Research, Taipei Veterans General Hospital, Taipei, Taiwan, Republic of China.
authors.forename: Anya Maan-Yuh
authors.initials: AM
authors.lastname: Lin
authors.affiliation: Department and Institute of Pharmacology, National Yang-Ming University.
authors.forename: Yu-Li
authors.initials: YL
authors.lastname: Lo
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000378:metabolism; D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002470:Cell Survival; D018432:Drug Resistance, Multiple / Q000187:drug effects*; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D015251:Epirubicin / Q000008:administration & dosage*; D058750:Epithelial-Mesenchymal Transition / Q000187:drug effects; D005260:Female; D006367:HeLa Cells / Q000187:drug effects; D064451:Hepcidins / Q000737:chemistry*; D006801:Humans; D008081:Liposomes / Q000737:chemistry*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D046529:Microscopy, Electron, Transmission; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000737:chemistry; D015398:Signal Transduction; D017210:Tilapia; D002583:Uterine Cervical Neoplasms / Q000378:metabolism / Q000473:pathology
chemical_list: D018435:ATP Binding Cassette Transporter, Subfamily B; D000276:Adjuvants, Immunologic; D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D053148:Caspase 3; D053453:Caspase 9; D015251:Epirubicin; D064451:Hepcidins; D008081:Liposomes; D000089762:Multidrug Resistance-Associated Protein 2; D027425:ATP-Binding Cassette, Sub-Family C Proteins; D017382:Reactive Oxygen Species; C513055:ABCB1 protein, human; C000715087:ABCC2 protein, human; C505407:CASP3 protein, human; C506433:CASP9 protein, human; C421109:multidrug resistance-associated protein 1
keywords: antimicrobial peptide; apoptosis; autophagy; epirubicin; liposomes; multidrug resistance