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The activation pathway of nuclear transcription factor kappa B (NF-κB) is a key mechanism for the progression of carcinogenesis at the molecular level. NF-κB is related to the promotion of cell proliferation, inhibition of apoptosis, and the enhancement of tumor metastasis and angiogenesis. Marasmius oreades culture liquid extract, which was previously shown to affect NF-κB activation through inhibition of the phosphorylation of the inhibitory protein kappa B (IκBα), was subjected to liquid chromatography in order to investigate the specific mechanism of action of the active moieties present in the extract. Four fractions were obtained and tested for their abilities to block NF-κB activation pathway at different molecular levels. All fractions showed an anti-proliferative potential with no apparent cytotoxicity on MCF7 breast cancer cell line. Two out of the four fractions strongly affected the phosphorylation of IκBα …