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

Chaogang Tong,Zhiwen Feng,Jun Zhao,Bin Cheng,Xiangming Wang,Dafei Dai,Xiaopeng Chen,Yuhai Wu

doi:10.1038/s41420-021-00411-5

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.

Highlights

Hepatocellular carcinoma (HCC) is a kind of high malignant tumors with an extremely poor prognosis[1,2], while intrahepatic metastasis or recurrence is the main cause to be blamed for this situation
The results revealed that HPSE expression levels were markedly upregulated in HCC tissues (Fig. 1a, b)
The HPSE mRNA and protein expression levels were markedly decreased (Fig. 2a, b). It was found in the following trans-endothelial migration (TEM) experiment that the TEM rate of HCC cells in the shHPSE group was significantly lower than that in the shCtrl group (Fig. 2c). These results suggest that HPSE promotes the invasion and migration of HCC cells

Summary

Introduction

Hepatocellular carcinoma (HCC) is a kind of high malignant tumors with an extremely poor prognosis[1,2], while intrahepatic metastasis or recurrence is the main cause to be blamed for this situation. Heparanase (HPSE) is a kind of multifunctional extracellular hydrolase, which is up-regulated in almost all human malignant tumors[4]. As the hydrolytic substrate of HPSE, heparan sulfate proteoglycans (HSPGs), composed by a core protein with covalently attached heparan sulfate (HS) side chains, are widely distributed in the extracellular matrix (ECM) and cell surface[5]. The HS side chains are able to bind to a variety of proteins, including basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), and tumor necrosis factor-α (TNFα), providing a local extracellular storage depot in various tissues[5]. Based on the property of HS side chain, HPSE promotes tumor growth, metastasis, and angiogenesis by degrading HSPGs and releasing VEGF and bFGF from the extracellular cytokines depot[6]

Methods

Results

Conclusion