Evaluation of anti-cancer effects of carnosine and melittin-loaded niosomes in MCF-7 and MDA-MB-231 breast cancer cells

Abstract

Background: We investigated the anti-cancer effect of carnosine-loaded niosomes (Car-NIO) and melittin-loaded niosomes (Mel-NIO) with olaparib in breast cancer cell lines (MCF-7 and MDA-MB-231). Methods: The thin film method was used for preparing the niosomes and characterized in terms of morphology, size, and polydispersity index (PDI). We further evaluated the impact of these peptides on breast cancer cells viability, RT-qPCR assays, malondialdehyde (MDA) activity, and cell cycle progression, to determine if these are linked to carnosine and melittin's anti-proliferative properties. Results: Car-NIO and Mel-NIO in vitro study inhibited cancer cell viability. They have also upregulated the expression of protein 53 (P53), BCL2-Associated X Protein (Bax), caspase-9, caspase-3, programmed cell death 4 (PDCD4), and Forkhead box O3 (FOXO3), while downregulated the expression of B-cell lymphoma 2 (Bcl2), poly (ADP-ribose) polymerase (PARP 1), and MicroRNA-183 (miRNA-183). The MCF-7 cells were arrested at the G2/M phase in Car-NIO, on the other hand, the MDA-MB-231 cells were arrested at the S phase. While the Mel-NIO and olaparib arrested the MCF-7 and MDA-MB-231 cells at the G0/1 phase. Conclusion: Our study successfully declared that Mel-NIO had more anti-cancer effects than Car-NIO in both MCF-7 and MDA-MB-231 breast cancer cells.

Keywords: breast cancer cells; carnosine; cell cycle analysis; melittin; miRNA-183; niosome.

FIGURE 1

Characterization of Car-NIO and Mel-NIO. Dynamic light scattering size measurement of Car-NIO (A). TEM image of the Car-NIO (B). Mel-NIO dynamic light scattering size measurement (C). TEM image of the Mel-NIO (D).

FIGURE 2

In vitro drug release profile of carnosine and Car-NIO (A), melittin and Mel-NIO (B) from the dialysis bag in pH 5.4 at 37°C. Results are represented by mean ± SD (n = 3). *p < 0.05, **p < 0.001.

FIGURE 3

Comparing stability of optimum formulation at 4°C and 25°C of Car-NIO. Mean particle size (A), PDI (B) and EE % (C) were studied as stability parameters. Results are represented by mean ± SD (n = 3). *p < 0.05, **p < 0.001.

FIGURE 4

Comparing stability of optimum formulation at 4°C and 25°C of Mel-NIO. Mean particle size (A), PDI (B) and EE % (C) were studied as stability parameters. Results are represented by mean ± SD (n = 3). *p < 0.05, **p < 0.001.

FIGURE 5

The impact of different concentrations of Car-NIO, Mel-NIO, and olaparib on the viability of MCF-7 (A), and MDA-MB-231 (B) cells by using the MTT assay after 48 h of incubation. Data are shown as mean ± SEM, and p < 0.05.

FIGURE 6

The impact of Car-NIO, Mel-NIO and olaparib on the genes expressions levels of P53 (A), Bax (B), caspase-9 (C), and caspase-3 (D) (relative gene mRNA expressions/GADPH) in the MCF-7 and MDA-MB-231 cell lines. * Significant difference between control and treated group (p < 0.05), ** highly significant difference between control and treated group (p < 0.01).

FIGURE 7

The impact of Car-NIO, Mel-NIO and olaparib on the genes expressions levels of Bcl2 (A), PARP 1 (B), MicroRNA-183 (C), PDCD4 (D) and FOXO3 (E) (Relative gene mRNA expressions/GADPH) in the MCF-7 and MDA-MB-231 cell lines. * Significant difference between control and treated group (p < 0.05), ** highly significant difference between control and treated group (p < 0.01).

FIGURE 8

The impact of Car-NIO, Mel-NIO, and olaparib on the level of MDA (nmol/mg) in the MCF-7 and MDA-MB-231 cell lines. * Significant difference between control and treated group (p < 0.05), ** highly significant difference between control and treated group (p < 0.01).

FIGURE 9

The impact of Car-NIO, Mel-NIO, and olaparib on cell cycle progression of breast cancer MCF-7 (A, B), and MDA-MB-231 (C, D) cell lines. * Significant difference between control and treated group (p < 0.05), ** highly significant difference between control and treated group (p < 0.01).