Heparanase induces necroptosis of microvascular endothelial cells to promote the metastasis of hepatocellular carcinoma

Abstract

Heparanase (HPSE) is a kind of multifunctional extracellular hydrolase, and related to metastasis of hepatocellular carcinoma (HCC). Endothelial necroptosis promotes the metastasis of cancer cells. It is not clear whether HPSE could mediate necroptosis of microvascular endothelial cells (MVECs) to promote HCC metastasis. Here we found HPSE expression was up-regulated in HCC tissues and its over-expression was correlated with multiple tumor foci, microvascular invasion, and poor outcome of HCC patients. Non-contact co-culture experiments showed high-expressed HPSE in HCC cells mediated the necroptosis of human umbilical vein endothelial cells (HUVECs) and elevated the expression levels of syndecan-1 (SDC-1) and tumor necrosis factor-α (TNF-α) in vitro. As a result of necroptosis, trans-endothelial migration (TEM) of HCC cells was increased. Conversely, both HPSE and SDC-1 knockdowns reversed necroptosis and decreased TNF-α expression level, while HPSE over-expression increased SDC-1 and TNF-α expression and aggravated necroptosis. Animal experiments found that the nude mice, intraperitoneally injected with HPSE high expressing HCC cells, had obvious necroptosis of MVECs and high intrahepatic metastasis rate, which could be relieved by inhibitor of necroptosis. Morever, HPSE elevated the expression levels of p38 mitogen-activated protein kinase (p38 MAPK) rather than nuclear factor kappa B in vitro. Our data suggest that HPSE induces necroptosis of MVECs to promote the metastasis of HCC by activating HPSE/SDC-1/TNF-α axis and p38 MAPK pathway.

The Heparanase Regulatory Network in Health and Disease

“…Transwell co-culture of heparanase-silenced hepatocellular carcinoma (HCC) cells with HUVECs protected HUVECs from MLKL mRNA and protein upregulation and necroptosis In transwell co-cultures of heparanase-overexpressing HCC cells and HUVECs, HUVECs displayed higher MLKL protein expression after co-culture compared to controls Necroptosis [191] matrix metalloprotease-2 (MMP-2)…”

“…In transwell co-cultures of heparanase-silenced HCC cells and HUVECs, HUVECs displayed lower p38 mRNA and phosphorylated protein expression after co-culture compared to controls Necroptosis [191] PDK2 Nuclear heparanase regulated the mRNA expression of PDK2 through HAT activation.…”

“…In transwell co-cultures of heparanase-overexpressing HCC cells and HUVECs, HUVECs displayed higher RIPK1 and RIPK3 protein expression after co-culture compared to controls Necroptosis [191] SDF-1 (CXCL-12) SDF-1 expression was reduced in heparanase-deficient macrophages. Heparanase regulated SDF-1 expression through Erk, p38, and JNK signaling pathway…”

“…Inhibiting heparanase with either PG545 or PI-88 in patient-derived explants of normal mammary tissue reduced syndecan-1 mRNA expression Tissue density and breast cancer [192] In transwell co-cultures of heparanase-silenced HCC cells and HUVECs, HUVECs displayed lower syndecan-1 mRNA and protein expression after co-culture compared to controls. In transwell co-cultures of heparanase-overexpressing HCC cells and HUVECs, HUVECs displayed higher syndecan-1 mRNA and protein expression after co-culture compared to controls Necroptosis [191] Tissue factor (TF) mRNA expression levels of TF were elevated in heparanase transfected breast carcinoma cells and transgenic mice over-expressing heparanase. Exogenous addition of heparanase also induced TF expression in human promyelocytic leukemia cells.…”

“…Pre-treatment with heparanase inhibitior PG545 reduced the expression of TNF-α EMT/Acute kidney injury [179] Human melanoma cells deficient in heparanase exhibited increased TNF-α expression Melanoma progression [169] Heparanase upregulated the expression of TNF-α in PMA-activated U937 macrophage cells. Treatment of cells with heparanase inhibitor SST0001 reduced TNF-α expression Acute kidney injury/M1 macrophage polarization [182] HS fragments generated by heparanase activated TLR4, MyD88, and NF-κB to upregulate TNF-α Inflammation [152] TNF-α mRNA expression was reduced in chemotherapy-treated macrophages isolated from heparanase knockout mice In transwell co-cultures of heparanase-silenced HCC cells and HUVECs, HUVECs displayed lower TNF-α mRNA and protein expression compared to controls Tumor Growth/Induction of pro-inflammatory cytokine expression by chemotherapy-treated macrophage [123] In transwell co-cultures of heparanase-overexpressing HCC cells and HUVECs, HUVECs displayed higher TNF-α mRNA and protein expression after co-culture compared to controls Necroptosis [191] Table 3. Cont.…”