Highlights Abstract β-glucans are polysaccharides comprising β-D-glucoses with various bioactivities. Herein, we extracted three β-glucans from Lentinus edodes with different sources and assessed their antitumor activities on a mice model with intragastric, intraperitoneal and intratumoral injection. Three polysaccharides were shown to have the same chemical structure of β-(1,3)-glucan with β-(1,6) branches, and