Abstract
BackgroundLentinus edodes is a medicinal mushroom widely used in Asian countries for protecting people against some types of cancer and other diseases.ObjectiveThe objective of the present study was to investigate the direct antiproliferation activity and the antitumor mechanisms of water-extracted polysaccharide (WEP1) purified from L. edodes in H22 cells and H22-bearing mice.DesignThe extraction, isolation, purification, and structure determination of the water-soluted L. edodes polysaccharide WEP1 were performed. The growth inhibitory effects of WEP1 on H22 cells and H22-bearing mice were determined by 3-(4,5-Dimethylthiazol-2-Yl)-2,5-Diphenyltetrazolium Bromide (MTT) method and animal studies. Flow cytometry, scanning electron microscopy, and laser scanning confocal microscopy were used to observe the morphological characteristics of apoptotic cells. The levels of intracellular reactive oxygen species (ROS) were detected by flow cytometry using 2’,7’-dichlorofluorescein-3’,6’-diacetate (DCFH-DA). Western blot was used to determine the expressions of cell cycle proteins and apoptosis-related proteins.ResultsResults showed that WEP1 with a molecular weight of 662.1 kDa exhibited direct antiproliferation activity on H22 cells in a dose-dependent manner. In vivo, WEP1 significantly inhibited the growth of tumor at different doses (50, 100, and 200 mg/kg) and the inhibition rates were 28.27, 35.17, and 51.72%, respectively. Furthermore, morphological changes of apoptosis and ROS overproduction were observed in H22 cells by WEP1 treatment. Cell cycle assay and western blot analyses indicated that the apoptosis induction activity of WEP1 was associated with arresting cell cycle at G2/M phase and activating mitochondrial-apoptotic pathway. Besides, WEP1 disrupted the microtubule network accompanied by alteration of cellular morphology.ConclusionResults suggested that the antitumor mechanisms of WEP1 might be related to arresting cell cycle at G2/M phase, inhibiting tubulin polymerization and inducing mitochondrial apoptosis. Therefore, WEP1 possibly could be used as a promising functional food for preventing or treating liver cancer.
Highlights
Lentinus edodes is a medicinal mushroom widely used in Asian countries for protecting people against some types of cancer and other diseases
To elucidate the inhibitory effect of WEP1 on H22 cells, cells were treated with various concentrations of WEP1 (12.5, 25, 50, 100, 200, 400, and 800 μg/mL) for 24 h in vitro, and followed by MTT assay
Results indicated that WEP1 showed obvious antitumor properties at different concentrations, with the inhibition ratios being 28.27, 35.17, and 51.72%, respectively, and the antitumor effect of the high-dose WEP1 group was comparable to those of CTX (51.72% vs. 54.48%, Table 1), which was commonly used as a chemotherapy drug in clinical
Summary
Lentinus edodes is a medicinal mushroom widely used in Asian countries for protecting people against some types of cancer and other diseases. Objective: The objective of the present study was to investigate the direct antiproliferation activity and the antitumor mechanisms of water-extracted polysaccharide (WEP1) purified from L. edodes in H22 cells and H22-bearing mice. Design: The extraction, isolation, purification, and structure determination of the water-soluted L. edodes polysaccharide WEP1 were performed. Western blot was used to determine the expressions of cell cycle proteins and apoptosis-related proteins. Morphological changes of apoptosis and ROS overproduction were observed in H22 cells by WEP1 treatment. Cell cycle assay and western blot analyses indicated that the apoptosis induction activity of WEP1 was associated with arresting cell cycle at G2/M phase and activating mitochondrial-apoptotic pathway. Conclusion: Results suggested that the antitumor mechanisms of WEP1 might be related to arresting cell cycle at G2/M phase, inhibiting tubulin polymerization and inducing mitochondrial apoptosis. WEP1 possibly could be used as a promising functional food for preventing or treating liver cancer
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