Ganoderma lucidum Spore Polysaccharide Inhibits the Growth of Hepatocellular Carcinoma Cells by Altering Macrophage Polarity and Induction of Apoptosis

Abstract

Background. Ganoderma lucidum has certain components with known pharmacological effects, including strengthening immunity and anti-inflammatory activity. G. lucidum seeds inherit all its biological characteristics. G. lucidum spore polysaccharide (GLSP) is the main active ingredient to enhance these effects. However, its specific biological mechanisms are not exact. Our research is aimed at revealing the specific biological mechanism of GLSP to enhance immunity and inhibit the growth of H22 hepatocellular carcinoma cells. Methods. We extracted primary macrophages (Mø) from BALB/c mice and treated them with GLSP (800 μg/mL, 400 μg/mL, and 200 μg/mL) to observe its effects on macrophage polarization and cytokine secretion. We used GLSP and GLSP-intervened macrophage supernatant to treat H22 tumor cells and observed their effects using MTT and flow cytometry. Moreover, real-time fluorescent quantitative PCR and western blotting were used to observe the effect of GLSP-intervened macrophage supernatant on the PI3K/AKT and mitochondrial apoptosis pathways. Results. In this study, GLSP promoted the polarization of primary macrophages to M1 type and the upregulation of some cytokines such as TNF-α, IL-1β, IL-6, and TGF-β1. The MTT assay revealed that GLSP+Mø at 400 μg/mL and 800 μg/mL significantly inhibited H22 cell proliferation in a dose-dependent manner. Flow cytometry analysis revealed that GLSP+Mø induced apoptosis and cell cycle arrest at the G2/M phase, associated with the expression of critical genes and proteins (PI3K, p-AKT, BCL-2, BAX, and caspase-9) that regulate the PI3K/AKT pathway and apoptosis. GLSP reshapes the tumor microenvironment by activating macrophages, promotes the polarization of primary macrophages to M1 type, and promotes the secretion of various inflammatory factors and cytokines. Conclusion. Therefore, as a natural nutrient, GLSP is a potential agent in hepatocellular carcinoma cell treatment and induction of apoptosis.

Drug Resistance in Colorectal Cancer: From Mechanism to Clinic

“…TP-16 (EP4 antagonist) treatment polarized TAM from immunosuppressive M2 type to pro-inflammatory M1 type, which enhanced T-cell-mediated anti-tumor immunity and improved the efficacy of anti-PD-1 [ 58 ]. In addition, a series of studies have proved that Ganoderma lucidum spore polysaccharide (GLSP) [ 337 ], CMPB90-1 [ 338 ], natural M1 exosomes [ 339 ], conditional deletion of the microRNA (miRNA)-processing enzyme DICER [ 334 ] and TLR 7/8 agonist [ 57 ] can specifically eliminate M2-like phenotype or reset TAM from M2-like phenotype to M1-like phenotype to inhibit tumor development and reverse tumor treatment resistance.…”

The Regulatory Roles of Polysaccharides and Ferroptosis-Related Phytochemicals in Liver Diseases

“…[ 78 , 79 ]. Ganoderma lucidum spore polysaccharide (GLSP) promoted the polarization of primary macrophages into M1 type and cytokine expression (such as TNF-α, IL-1β, IL-6, and TGF-β1) [ 80 ].…”

“…APS, GLSP, fucoidan, Pleurotus ostreatus polysaccharide, ginger polysaccharide, Aconitum coreanum polysaccharide, pumpkin polysaccharide (PPPF), Rhizopus Nigrum polysaccharide, and an acid-soluble polysaccharide from Grifola frondose could inhibit hepatocellular carcinoma growth by apoptosis [ 78 , 79 , 80 , 85 , 86 , 87 , 88 , 89 , 90 ]. The JAK/STAT, PI3K/AKT, and RAS/ERKs pathways are enhanced in many HCC cells, conferring on them resistance to apoptotic stimuli [ 91 ].…”

“…The JAK/STAT, PI3K/AKT, and RAS/ERKs pathways are enhanced in many HCC cells, conferring on them resistance to apoptotic stimuli [ 91 ]. GLSP triggers HCC cell apoptosis via regulating the PI3K/AKT pathway, with increased Bax/caspases and decreased Bcl-2 [ 80 ]. PPPF treatment induces apoptosis in HepG2 cells by increasing the protein tyrosine phosphatase SHP-1 to inhibit JAK2/STAT3 signaling [ 88 ].…”

Research Progress on the Anticancer Activities and Mechanisms of Polysaccharides From Ganoderma

“…However, the exact mechanism responsible for GLP-associated reduction in CD68 + macrophages needs to be illuminated in future studies. It was reported that GLP fostered primary macrophage polarization to M1 type and increased the production of TGF-β1, TNF-α, IL-1β, and IL-6 ( Song et al, 2021 ). GLP-intervened macrophages evidently repressed HCC cell proliferation by inducing cell cycle arrest and activating the mitochondrial apoptosis pathway via the PI3K/Akt signaling cascade.…”