The Use of a Liposomal Formulation Incorporating an Antimicrobial Peptide from Tilapia as a New Adjuvant to Epirubicin in Human Squamous Cell Carcinoma and Pluripotent Testicular Embryonic Carcinoma Cells

Abstract

This study aims to explore the effects and mechanisms of hepcidin, a potential antimicrobial peptide from Tilapia, and epirubicin (Epi), an antineoplastic agent, on the generation of reactive oxygen species (ROS) and link the ROS levels to the reversal mechanisms of multidrug resistance (MDR) by epirubicin and hepcidin in human squamous cell carcinoma SCC15 and human embryonal carcinoma NT2D1 cells. The cells, pretreated with hepcidin, epirubicin, or a combination of these compounds in PEGylated liposomes, were used to validate the molecular mechanisms involved in inhibiting efflux transporters and inducing apoptosis as evaluated by cytotoxicity, intracellular accumulation, mRNA levels, cell cycle distribution, and caspase activity of this combination. We found that hepcidin significantly enhanced the cytotoxicity of epirubicin in liposomes. The co-incubation of epirubicin with hepcidin in liposomes intensified the ROS production, including hydrogen peroxide and superoxide free radicals. Hepcidin significantly increased epirubicin intracellular uptake into NT2D1 and SCC15 cells, as supported by the diminished mRNA expressions of MDR1, MDR-associated protein (MRP) 1, and MRP2. Hepcidin and/or epirubicin in liposomes triggered apoptosis, as verified by the reduced mitochondrial membrane potential, increased sub-G1 phase of cell cycle, incremental populations of apoptosis using annexin V/PI assay, and chromatin condensation. As far as we know, this is the first example showing that PEGylated liposomal TH1-5 and epirubicin gives rise to cell death in human squamous carcinoma and testicular embryonic carcinoma cells through the reduced epirubicin efflux via ROS-mediated suppression of P-gp and MRPs and concomitant initiation of mitochondrial apoptosis pathway. Hence, hepcidin in PEGylated liposomes may function as an adjuvant to anticancer drugs, thus demonstrating a novel strategy for reversing MDR.

Induction of apoptosis in response to improved gedunin by liposomal nano-encapsulation in human non-small-cell lung cancer (NCI-H292) cell line

“…As shown in Table 1 the encapsulation efficiency of liposomes (86.82%) was as high as those produced by other researchers [22] , but the ones used in the present study were of larger size distribution (394 nm), and had an anionic zeta potential (-54) in contrast to their cationic liposomes. Unlike anionic liposomes, cationic liposomes cause induction of cellular stress and change in gene expression [22]. Most of the phosphatidylcholines usually used for liposomal drug delivery have been proven as non-toxic [5,6].…”

Cationic PEGylated liposomes incorporating an antimicrobial peptide tilapia hepcidin 2–3: an adjuvant of epirubicin to overcome multidrug resistance in cervical cancer cells

“…5 The combined treatment of hepcidin 1-5 and epirubicin reduced cell growth in human squamous carcinoma and testicular carcinoma cells via ROS-mediated inhibition of MDR transporters and triggering of mitochondrial apoptosis pathway. 8 Resistance to apoptotic signaling and overexpression of ABC transporters in cancer cells considerably reduced the effectiveness of cancer chemotherapy. 51 Based on our current results, we suggest that PEGylated liposomal epirubicin and hepcidin 2-3 formulations might affect the levels of ROS and thus coordinately regulate transporter-, apoptosis-and/or autophagy-related mechanisms, thereby inducing MDR reversal.…”

“…4 Interestingly, our previous investigation has also verified that tilapia hepcidin 1-5 and epirubicin caused cell death in human squamous carcinoma and testicular embryonic carcinoma cells through the suppression of drug efflux pumps and the simultaneous activation of mitochondrial apoptosis pathway. 8 Nevertheless, the possibility of hepcidin 2-3 as an adjuvant to potentiate the activity of anticancer drugs has not been addressed in the aforementioned reports.…”

“…25,26 Epirubicin, an anticancer drug of anthracycline, exhibits a potent apoptosis-triggering effect via the intrinsic mitochondrial signaling in various cancer types. 8,13,27 It is also a substrate of P-gp, MRP1, and MRP2. 28,29 Due to the facts that epirubicin may trigger oxidative stress and that the produced reactive oxygen species (ROS) are potential cell death modulators, 30,31 evaluation of the role of autophagy, necroptosis, and/or apoptosis in cytotoxicity induced by cotreatment of epirubicin and hepcidin 2-3 in HeLa cells is critical.…”

Use of Tilapia Hepcidin in Oral Cancer Therapeutics: A Proposal

“…More recently, nanotechnologists have incorporated TH into liposomal formulations and enhanced the therapeutic effect of "epirubicin" on human tongue squamous cell carcinoma (SCC-15), human embryonal carcinoma cells, and Hela cells. 11,12 It led to a rise in cell death through reduced epirubicin efflux via reactive oxygen species (ROS)-mediated suppression and concomitant initiation of the mitochondrial apoptosis pathway. Thus, TH affects these cells by suppressing the multidrug resistance transporters, apoptosis, autophagy, and/or necroptosis.…”