Metabolic profiling and systems biology analysis of Flammulina filiformis Fv19 extract: an integrated computational and experimental approach for dual anti-cancer and immunomodulatory therapy

Abstract

BACKGROUND: The increasing demand for multi-target cancer therapies has prompted investigation into the anticancer and immunomodulatory potential of the hot water extract of Flammulina filiformis Fv19, utilizing in vitro cytotoxicity assays, in vivo murine models, and molecular docking analyses. RESULTS: The hot water extraction of mushroom mycelia yielded 11.4 ± 1.2% (w/w). Gas chromatography-mass spectrometry (GC-MS) profiling identified six major metabolites, with the most abundant being cholestan3ol, 2methylene (3α, 5α) (37.47%); spirost8en11one, 3hydroxy (23.24%); and 1Hcycloprop[e]azulen7ol, decahydro1,1,7trimethyl4methylene (12.58%). The extract demonstrated potent inhibition of MCF-7 (IC₅₀ = 5 µg/ml) and Hep-G2 (IC₅₀ = 4 µg/ml) cancer cell lines. In vivo evaluation involved male albino mice receiving oral administration of the extract at a dosage of 200 mg/kg/day for 10 days, employed in both preventive and treatment protocols against the Ehrlich solid tumor model. In vivo oral administration significantly suppressed Ehrlich carcinoma tumor growth with a 99.79% inhibition ratio, achieving complete tumor regression in 66.7% of mice. Additionally, the extract enhanced immune responses by increasing macrophage count by 166%, TNF-α production by 284%, NO release by 71%, and H₂O₂ levels by 55% compared to the tumor-bearing group, while reducing IL-17 production by splenocytes by 27%, indicating immunomodulatory effects. Molecular docking revealed strong interactions with oncogenic and immune-regulatory proteins, including Bcl-2 (-5.015 kcal/mol), EGFR (-4.567 kcal/mol), COX-2 (-4.581 kcal/mol), and iNOS (-5.855 kcal/mol). CONCLUSIONS: These findings highlight F. filiformis Fv19 extract as a promising natural anticancer and immunomodulatory agent for multi-target cancer therapy. CLINICAL TRIAL NUMBER: Not applicable.