Delivery of Melittin as a Lytic Agent via Graphene Nanoparticles as Carriers to Breast Cancer Cells

Abstract

Melittin, as an agent to lyse biological membranes, may be a promising therapeutic agent in the treatment of cancer. However, because of its nonspecific actions, there is a need to use a delivery method. The conducted research determined whether carbon nanoparticles, such as graphene and graphene oxide, could be carriers for melittin to breast cancer cells. The studies included the analysis of intracellular pH, the potential of cell membranes, the type of cellular transport, and the expression of receptor proteins. By measuring the particle size, zeta potential, and FT-IT analysis, we found that the investigated nanoparticles are connected by electrostatic interactions. The level of melittin encapsulation with graphene was 86%, while with graphene oxide it was 78%. A decrease in pHi was observed for all cell lines after administration of melittin and its complex with graphene. The decrease in membrane polarization was demonstrated for all lines treated with melittin and its complex with graphene and after exposure to the complex of melittin with graphene oxide for the MDA-MB-231 and HFFF2 lines. The results showed that the investigated melittin complexes and the melittin itself act differently on different cell lines (MDA-MB-231 and MCF-7). It has been shown that in MDA-MD-231 cells, melittin in a complex with graphene is transported to cells via caveolin-dependent endocytosis. On the other hand, the melittin–graphene oxide complex can reach breast cancer cells through various types of transport. Other differences in protein expression changes were also observed for tumor lines after exposure to melittin and complexes.

Effect of Melittin Complexes with Graphene and Graphene Oxide on Triple-Negative Breast Cancer Tumors Grown on Chicken Embryo Chorioallantoic Membrane

“…It is worth emphasizing that the effects of the studied complexes are the result of simultaneous exposure to the nanomaterial and M and their interaction as factors harmful to cancer cells. The use of GN and GO allows the liberation of M by the nanomaterial, which has been confirmed in previous studies [42], and the reduction of its spread outside the tumor when administered into the tumor. Oxidative stress can cause the production of cytokines, which in turn are used to communicate with other cells [77].…”

“…The level of encapsulation of M with GN was 86% and with GO it was 78%, which is a suitable indicator for potential therapeutic agents. It has been proven that MGN is transported to MDA-MB-231 cells through caveolin-dependent endocytosis, while MGO is transported in multiple ways [42]. In addition, the use of carbon nanoparticles reduced the rate of cell death by necrosis to a greater extent compared to the reduction resulting from M alone [22].…”

“…The use of GN nanoparticles as carriers did not reduce toxicity to healthy cells; however, these investigations were in vitro studies with a monoculture. In addition, the results of previous studies indicate the potential use of GN and GO as carriers due to their electrostatic interaction with M and the high degree of M entrapment efficiency [42]. In this research, the response of tumors cultured on chicken embryo chorioallantoic membrane (CAM) to treatment with the tested complexes and to M alone was examined.…”

The current landscape of the antimicrobial peptide melittin and its therapeutic potential

“…Melittin-DM1 significantly increased the survival rates of melanoma mice than those under DM1 management alone. Moreover, a series of studies ( 86 , 87 , 179 ) investigated several strategies, such as innocuous nonspecific toxicity and increased stability of melittin in anti-breast cancer therapy. Daniluk and colleagues showed that compared to melittin alone, using diamond, pristine graphene, graphene oxide and carbon nanoparticles as the carriers for melittin reduced the level of necrosis in MDA-MD-231 and MCF-7 cells ( 86 ).…”

“…The selectivity of melittin to malignant breast cancer cells has attracted much attention ( 85 ). The tumoricidal properties of melittin on breast cancer cells have been investigated ( 86 , 87 ). Melittin could impede primary tumor growth and inhibit the growth of the metastatic sentinel lymph node in breast cancer models in vitro and in vivo ( 88 ).…”