Apoptosis induction activity of polysaccharide from Lentinus edodes in H22-bearing mice through ROS-mediated mitochondrial pathway and inhibition of tubulin polymerization

Abstract

Background 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. 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. Results Results 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. 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. Therefore, WEP1 possibly could be used as a promising functional food for preventing or treating liver cancer.

Physicochemical characterization of a polysaccharide from Rosa roxburghii Tratt fruit and its antitumor activity by activating ROS mediated pathways

“…Apoptosis is programmed cell death, and ROS is critical for promoting apoptosis in response to various apoptotic stimuli ( Khan et al, 2019 ; Yang et al, 2018 ). Induction of apoptosis and targeting ROS through polysaccharide intervention is an effective way to combat tumors ( Zhang et al, 2020 , 2021 ). In this study, RTFP-1 exhibited antiproliferative activity against HepG2 cells through inducing cell apoptosis.…”

Preparation, Characterization, and In Vitro Release of Curcumin-Loaded IRMOF-10 Nanoparticles and Investigation of Their Pro-Apoptotic Effects on Human Hepatoma HepG2 Cells

“…Many studies have proven that ROS are closely related to mitochondria. ROS can directly damage the integrity of the mitochondrial membrane by hyperpolarizing the mitochondrial membrane and releasing cytochrome c into the cytoplasm [53,54]. In addition, ROS can also damage mitochondrial DNA, which is related to the respiratory chain, thereby aggravating the production of ROS [55].…”

Inonotus obliquus Polysaccharides Inhibited Cellular Growth of NCI-H23 and A549 Lung Cancer Cells Through G0/G1 Cell Cycle Arrest and ROS Mediated Cell Death

“…Previous studies on anti-cancer agents considered generating high levels of intracellular ROS as a sign of induced apoptosis in the cancer cells (Simon et al, 2000;Kowaltowski et al, 2004;Lee et al, 2019;Li et al, 2020). Results of the present study showed that the cells incubated with the IOP at 50 and 100 μg/mL and stained with DCFH-DA exhibited significantly high concentrations of ROS in both cancerous cell lines in a dose-dependent manner in comparison to untreated cells.…”

“…Therefore, treatment with IOP could disturb the redox balance in NCI-H23 and A549 cells, and hence ROS may be a key mediator, leading to mitochondrial dysfunction, protein oxidation and DNA damage in cancer cells, followed by cell death or apoptosis induced by various anticancer drugs (Xiao et al, 2013;Sahayanathan et al, 2020). Many mushroom polysaccharides have also been shown to promotes production of ROS and induce apoptosis in human cancer cells (Zhang et al, 2020).…”

“…Annexin V binds phosphatidylserine exposed on the cell surface and is therefore considered a simple tool for detecting apoptosis (Vermes et al, 1995). Several previous studies used annexin V for the detection of apoptosis induced by β-Sitosterol, lobaplatin and morusin in A549 cells (Rajavel et al, 2018;Zhang et al, 2019;Wang et al, 2020).…”