Evaluation of the effect of induction of cell death by doxorubicin and rosmarinic acid loaded on protein nanoparticles in MCF-7 breast cancer cell line

Abstract

Breast cancer is the second most common cancer among women and the second leading cause of death in the world. Rosmarinic acid has been recognized as an efficient candidate for suppressing several cancer cells by interfering with tumor cell proliferation, reducing inflammation, and inducing apoptosis. Doxorubicin is also a common drug that is used in the chemotherapy of all types of cancer, however, due to its less biocompatibility, it attacks healthy cells in addition to cancer cells and causes many side effects for patients. In this study, in order to improve the anticancer effects of rosmarinic acid and reduce the side effects of doxorubicin by reducing the dose used, however targeting it inside the body on the breast cancer cell line, it is suggested to use protein nanoparticles such as zein as a drug delivery system. Material and Method: The intended formulations include free doxorubicin, doxorubicin and rosmarinic acid, free doxorubicin and nanoparticulated rosmarinic acid, and nanoparticle doxorubicin and rosmarinic acid were made, and the characteristics of the nanoparticles, including their size, encapsulation rate, and zeta potential, respectively, were determined by particle optical scattering technique and Ultrafiltration was determined. Then the release of the drug was studied and the effect of drug groups, whether free or loaded, on breast cancer cells was studied by MTT test and compared with the effect of free drug. Next, the fluorescent cells treated with drug groups were measured after half and three and a half hours of treatment and photographed with a confocal microscope. Next, the cells treated with drug groups were stained with DAPI staining and photographed. Results: Nanoparticles with an average size of 217 nm and a loading percentage of 85% for rosmarinic acid and 51% for doxorubicin were prepared. Cytotoxic tests revealed that DOX and RA loaded with Zein have a lower IC50 compared to their free form. The results of the drug release test showed that drugs are better released from nanoparticles in acidic conditions of cancer cells than healthy cells with neutral pH. Flow cytometry showed that the loading of drugs with Zein increases their accumulation inside the cell and with the continuous release of drugs, they lead to increased cell death. DAPI staining also showed that nuclear fragmentation in apoptotic cells is more and more intense when the nanoformulated drug is used.