Investigating the apoptotic and antimetastatic effect of daphnetin-containing nano niosomes on MCF-7 cells

Abstract

Breast cancer is the most common cancer in the world and the second leading cause of cancer-related deaths in women worldwide. Although great progress has been made in elucidating the molecular features and underlying pathogenesis of breast tumors and various therapeutic strategies have been applied for individualized treatment, some types of breast cancer patients with aggressive features have a poor prognosis in terms of treatment. Nanotechnology is increasingly used in biology and medicine, including as a tool for diagnosing, treating and targeting tumors. The aim of this study is therefore to develop a drug delivery system based on niosomes loaded with daphnetin, a phytochemical coumarin. To confirm the synthesis of the loaded nano niosome, their physical and chemical properties were examined using SEM, FTIR and DLS. In this study, the toxic effect of daphnetin-loaded nanoparticles on the MCF-7 cell line was measured using the MTT assay. The expression level of apoptotic genes Bax and Caspase 3; and metastatic genes MMP2 and ITGA5 were quantitatively evaluated using the Real-time PCR method, and the division and metastatic potential of cancer cells were qualitatively evaluated by performing the scratch (repair) method. Finally, the effect of the investigated compounds on the amount of apoptosis and necrosis and the induced cell cycle was evaluated using the flow cytometry method.

The results of the SEM study showed that the synthesized nanoparticles had a spherical morphology and a diameter of less than 200 nm. The zeta potential was determined to be 39.1 mV using a DLS device. The results of the FTIR study also showed successful interactions between niosome and daphnetin. According to the flow cytometry results, the frequency of early apoptosis and delayed apoptosis was significantly higher in the cells treated with the IC₅₀ concentration of daphnetin-loaded nanoparticles than in the group treated with daphnetin and free niosome. The expression of apoptotic genes was also increased in the group treated with the IC₅₀ concentration of the loaded nanoparticles and the expression of antimetastatic genes was decreased. The results of the cell migration assay (scratch test) also show that treatment with the IC₅₀ concentration of the loaded nanoparticles can effectively control cell migration. Therefore, they can be considered as chemotherapeutic agents against breast cancer.