Phytochemical profiling, anticancer effects on hepatocellular carcinoma and antimicrobial activities of four edible fungi: Unraveling mechanisms by molecular docking studies

Edible mushrooms, with their nutritional composition, pleasant taste, and flavor, have been used in food and traditional medicine worldwide for many years. Due to their secondary metabolites, they exhibit broad biological activities. This research aimed to analyze the phenolic contents by Liquid chromatography-mass spectrometry (LC/MS) and investigate the anticancer and antimicrobial properties of Cantharellus cibarius, Craterellus cornucopioides, Ramaria formosa, and Hydnum repandum extracts. The antimicrobial and antioxidant activity of extracts was determined by the well diffusion method and colorimetric assay kit, respectively. The extracts' ability to inhibit cell proliferation was tested on hepatocellular carcinoma (HCC) cell line HepG2 and Human umbilical vein endothelial cells (HUVECs) using the WST-1 (2-(4-iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2Htetrazolium, monosodium salt) assay. The effect of H. repandum extract on the levels of Bcl-2-Associated X (Bax), B-cell lymphoma 2 (Bcl-2), caspase-3, cyclin D1, and p53 proteins' expression was assessed through the western blot, and its influence on apoptosis was investigated using flow cytometry. Antimicrobial activity results revealed that the extracts produced inhibition zones ranging from 9.8 to 16.8 mm against bacteria and 9.8-17.6 mm against yeasts. H. repandum showed high antibacterial and antioxidant (7.45 +/- 0.17 mmol Trolox equivalent/L) activities attributable to its high chlorogenic and gallic acid content. H. repandum also showed the highest selective cytotoxicity against HepG2 cells compared to control HUVECs. H. repandum extract triggered apoptosis in HepG2 cells, markedly elevated the expression of Bax, p53, and caspase-3, and substantially reduced the levels of cyclin D1. Binding interactions between caspase-3 and the most abundant compounds in the H. repandum extract, alongside their pharmacokinetic profile, were supported by molecular docking analysis and absorption, distribution, metabolism, and excretion (ADME) studies.