Phytochemical profiling and anticancer potential of pistachio wastes against MCF-7 breast cancer cells: a metabolic and apoptotic pathway analysis

Abstract

Food waste poses significant environmental, economic, and social challenges worldwide. Pistachio waste is recognized for its health benefits, particularly for its anticancer properties. This study assessed the phytochemical characteristics of pistachio hull (PH) and shell (PS) using various solvents (ethyl acetate, hexane, and methanol). Methanolic extracts showed substantially higher phenolic and flavonoid contents along with enhanced antioxidant activity across all tested methods, with PH consistently outperforming PS in antioxidant properties. Cell viability tests on MCF-7 cancer cells indicated that PH-Hexane had a slightly greater effect than PH-ethyl acetate and PH-methanol. Cell cycle analysis revealed a notable arrest in the G0/G1 phase compared to the control group. Additionally, PH-Hexane treatment resulted in 21.24% apoptosis, primarily early apoptosis at 19.41%. The pathway analysis involving 46 genes revealed that PH-Hexane induces apoptosis in MCF-7 cancer cells by modulating several metabolic pathways, particularly those associated with the immune system, developmental biology, signal transduction, and cellular responses to stimuli. This induction was characterized by the substantial upregulation of the CEBPB, HIF1A, and MYC genes and the significant downregulation of ATF2, CEBPA, and NR3C1 genes. The findings of this study indicate that the hexane extract derived from high-oil-content wastes has potential as an anticancer agent. Future research on oil-rich plant-based wastes should consider utilizing hexane as a solvent for further exploration of anticancer properties.

Keywords: Anticancer; Antioxidants; Flavonoids; Food supplements; Pistachio; Waste valorization.

Fig. 1

Total phenolics (TPC), flavonoids (TFC), anthocyanins (Tant), protein (Tprotein), and carbohydrates (Tcarb) according to the tissues and solvents. The data were expressed as mean ± standard deviation

Fig. 2

DPPH scavenging activity, effective concentration 50 (EC50), Cupric Reducing Antioxidant Capacity (CUPRAC), ABTS scavenging acitvity, and Ferric Reducing Antioxidant Power (FRAP) in different pistachio tissues by the solvents. The data were expressed as mean ± standard deviation

Fig. 3

Impact of various pistachio wastes and solvents on cell viability at 48 and 96 h. PH: pistachio hull, PS: pistachio shell

Fig. 4

Gene expression levels of PH-Hexane extracts

Fig. 5

Metabolic pathways significantly influenced by PH-Hexane extract treatment, based on the expression of 46 genes

Fig. 6

Main metabolic pathways in “Developmental biology” root-pathway. The blue color indicates the significance and proportions of entities (Developemantal Biology, Reactome, R-HSA-1266738.17)

Fig. 7

The regulation of Diseases of signal transduction by growth factor receptors and second messengers and Diseases of mitotic cell cycle metabolic ways by PH-Hexane treatment to MCF-7 cancer cell line. The illustration was created in Reactome according to [103]

Fig. 8

Modulations in signal transduction metabolic pathway as a result of PH-Hexane treatments to MCF-7 cancer cells. Illustration was created via Reactome according to [112]

Fig. 9

Transcriptions of sub pathways in Cytokine Signaling in Immune system by the PH-Hexane treatment in MCF-7 cancer cells. The illustration was creatied according to [120] in the Reactome database

Fig. 10

The cell cycle analysis of MCF-7 cancer cells under control and PH-Hexane treatments

Fig. 11

Apoptosis in MCF-7 cancer cells by the treatment of control (A) and PH-Hexane (B)