Asparagus Polysaccharide inhibits the Hypoxia-induced migration, invasion and angiogenesis of Hepatocellular Carcinoma Cells partly through regulating HIF1α/VEGF expression via MAPK and PI3K signaling pathway

Abstract

Aim: Although there are so many treatment strategies used for hepatocellular carcinoma (HCC), the overall survival (OS) of HCC patients still remains very low. In our previous studies, asparagus polysaccharide (ASP) has been demonstrated to suppress proliferation, migration, invasion and angiogenesis of HCC cells under normoxic conditions in vitro. However, the inhibitory effects of ASP on the hypoxia-induced migration, invasion and angiogenesis of HCC cells still remain largely unexplored. Materials and methods: Cell Counting Kit-8 (CCK-8) assay, transwell assay, and tube formation assay were used to determine the effects of ASP on hypoxia-induced proliferation, migration, invasion and angiogenesis of HCC cells. ELISA, Western blotting analysis and immunofluorescence assay were used to confirm the effects of ASP on the expressions of HIF-1α and VEGF at the protein level. Moreover, effects of ASP on signaling pathway-related proteins were investigated by Western blotting analysis. Immunohistochemistry (IHC) assay was applied to test the effects of ASP on angiogenesis-associated proteins of tumor cells. Results: We showed that ASP effectively suppressed hypoxia-induced proliferation, migration, invasion and angiogenesis of SK-Hep1 and Hep-3B cells in a dose-dependent manner. In addition, the inhibitory effect of ASP might be partly attributed to down-regulation of HIF1α and VEGF proteins in SK-Hep1 and Hep-3B cells under hypoxic conditions. Moreover, signaling pathway study indicated that ASP significantly down-regulated the hypoxia-induced expressions of p-AKT, p-mTOR and p-ERK, while it had little effects on AKT, mTOR and ERK. Besides, SK-Hep1 xenograft tumor models in nude mice further confirmed that the inhibitory effect of ASP on xenograft tumors might be exerted partly via down-regulation of HIF1α and VEGF through blocking MAPK and PI3K signaling pathways. Conclusions: Our findings suggested that ASP suppressed the hypoxia-induced migration, invasion and angiogenesis of HCC cells partly through regulating HIF-1α/VEGF expression via MAPK and PI3K signaling pathways.

Combining Network Pharmacology and Experimental Validation to Study the Action and Mechanism of Water extract of Asparagus Against Colorectal Cancer

“…The anticancer activity of polysaccharide, saponin, and flavonoid extracts from ASP has been reported in various cancers, such as lung and bladder cancer ( Bilušić et al, 2019 ), liver cancer ( Cheng et al, 2021 ), and ovarian cancer ( Xu et al, 2021 ). Some studies have found that asparagus saponin extracts have significant cytotoxic effects on colorectal cancer cells ( Jaramillo-Carmona et al, 2018 ), and the results of Bousserouel ( Bousserouel et al, 2013 ) and others have shown in more detail that methanolic extracts of ASP can induce apoptosis by activating the TRAIL death receptor pathway in SW480 and SW620 human colon adenocarcinoma cells.…”

“…ASP has nourishing, moistening, fire-clearing, cough-suppressing, and intestinal effects and exerts anti-inflammatory, antioxidant, anti-aging, immunity-enhancing, and anti-tumor pharmacological effect ( Samad et al, 2014 ; Lei et al, 2017 ; Cheng et al, 2019 ; Sun et al, 2020 ). Recent studies have reported that ASP polysaccharides can inhibit hypoxia-induced hepatocellular carcinoma cell migration, invasion, and angiogenesis by regulating hypoxia inducible factor alpha (HIF-1α) and vascular endothelial growth factor (VEGF) expression through mitogen activated protein kinase (MAPK) and phosphatidylinositol-4,5-bisphosphate 3-kinase (PI3K) signaling pathways ( Cheng et al, 2021 ). In addition, ASP-derived exosome-like nanovesicles could inhibit hepatocellular carcinoma cell proliferation, induce apoptosis, and inhibit tumor growth without significant side effects in mice ( Zhang et al, 2021 ).…”

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

“…Several polysaccharides could block HCC angiogenesis by downregulating hypoxia-inducible factor 1α (HIF-1α) and vascular endothelial growth factors (VEGFs). Moreover, asparagus and dandelion polysaccharides could inhibit MAPK and PI3K signaling pathways to block tumor angiogenesis [ 82 , 83 , 84 ].…”

Targeting mTOR Signaling by Dietary Polysaccharides in Cancer Prevention: Advances and Challenges

“…The researchers also observed a decrease in the protein levels of HIF-1α and its downstream vascular endothelial growth factor (VEGF) after ASP treatment. 133 Low molecular weight fucoidan (LMWF), extracted from brown algae, has been reported to inhibit tumor growth and capillary formation of tumors in bladder cancerbearing mice. Exposure of bladder cancer cells T24 to LMWF inhibited the accumulation of HIF-1 and the secretion of VEGF produced under hypoxia.…”