Two novel compounds, ergosterol and ergosta-5,8-dien-3-ol, from Termitomyces heimii Natarajan demonstrate promising antihepatocarcinoma activity

摘要

Background:Mushroom-derived components have immense potential to become a safe alternative in identifying lead anti-cancer molecules.Termitomyces heimii Natarajan(T.heimii)is a traditionally used edible mushroom with no previous record of anti-hepatocarcinoma activity.Methods:The anti-proliferative efficacy of the mushroom ethyl acetate extract was screened against a panel of seven cancer cell lines,namely Hep G2,MCF-7,MDA-MB-231,MAD-MB-436,MOLT-4,Reh,and K-562,and against peripheral blood mononuclear cells isolated from normal healthy donors by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide reduction assay.The impact of the extract on nuclear morphology was examined by 40,6-diamidino-2-phenylindole staining and the apoptotic potential of the extract was evaluated through flow cytometry and Annexin V-PI dual staining,followed by an in vitro scratch assay to elucidate the anti-migratory potential of the extract.The apoptotic and antimigratory effects were further validated using in silico molecular docking with four compounds,ergosterol,ergosta-5,8-dien-3-ol,lanosterol,and eburicol,against two anti-apoptotic proteins,Bcl-2 and Bcl-XL,and two angiogenic receptors,VEGFR-1 and VEGFR-2.Results:The screening data revealed that ethyl acetate extract exhibited remarkable anti-proliferative efficacy against Hep G2 cells,with a half maximal inhibitory concentration(IC50)value of 263.53(8.09)mg/mL,followed by MCF-7 cell lines and showed a negligible effect on peripheral blood mononuclear cells.A clear alteration of the cellular and nuclear morphology was concentration-dependently observed in Hep G2 cells.The extract induced robust apoptosis and a significant concentrationdependent increase in the scratch area.The results of in silico docking revealed that compared to standard drug sunitinib,both ergosterol and ergosta-5,8-dien-3-ol displayed lower binding energy,and satisfactory drugability and absorption,distribution,metabolism,excretion,and toxicity properties.Conclusions:T.heimii is a potential source for isolating lead anticancer molecules in the future.Ergosterol and ergosta-5,8-dien-3-ol hold great promise as new drugs against hepatocarcinoma.