Heparin Inhibits Hepatocyte Growth Factor Induced Motility and Invasion of Hepatocellular Carcinoma Cells through Early Growth Response Protein 1

Abstract

The Hepatocyte Growth Factor (HGF)/c-Met signaling pathway regulates hepatocyte proliferation, and pathway aberrations are implicated in the invasive and metastatic behaviors of hepatocellular carcinoma (HCC). In addition to c-Met, heparin acts as a co-receptor to modulate pathway activity. Recently, anti-metastatic and anti-cancer effects of heparin have been reported. However, the role of heparin in the regulation of HGF signaling remains controversial and the effects of heparin on HGF-induced biological responses during hepatocarcinogenesis is not yet defined. In this study we determined the effects of heparin on HGF-induced activities of HCC cells and the underlying molecular mechanisms. Here, we report for the first time that heparin inhibits HGF-induced adhesion, motility and invasion of HCC cells. In addition, heparin reduced HGF-induced activation of c-Met and MAPK in a dose-dependent manner, as well as decreased transcriptional activation and expression of Early growth response factor 1 (Egr1). HGF-induced MMP-2 and MMP-9 activation, and MT1-MMP expression, also were inhibited by heparin. Stable knockdown of Egr1 caused a significant decrease in HGF-induced invasion, as well as the activation and expression of MMPs. Parallel to these findings, the overexpression of Egr1 increased the invasiveness of HCC cells. Our results suggest that Egr1 activates HGF-induced cell invasion through the regulation of MMPs in HCC cells and heparin inhibits HGF-induced cellular invasion via the downregulation of Egr1. Therefore, heparin treatment might be a therapeutic approach to inhibit invasion and metastasis of HCC, especially for patients with active HGF/c-Met signaling.

A Mitogen-activated Protein Kinase/Extracellular Signal-regulated Kinase Kinase (MEK)-dependent Transcriptional Program Controls Activation of the Early Growth Response 1 (EGR1) Gene during Amino Acid Limitation

“…This observation is consistent with reports that EGR1 promotes the in vivo motility and invasion of hepatocellular carcinoma cells (45). Furthermore, one mechanism by which EGR1 may aid hepatocellular carcinoma proliferation in the face of nutrient limitation may be via induction of VEGF-A.…”

Heparin treatment increases thioredoxin interacting protein expression in hepatocellular carcinoma cells

“…In our previous studies, we showed the effects of heparin on hepatocyte growth factor induced motility and invasion in hepatocellular carcinoma (HCC) cells (Ozen et al, 2012). In order to identify the molecular mechanisms of heparin mediated biological responses, we performed a gene expression array with a HCC cell line.…”

“…Germany) as described previously (Ozen et al, 2012). Briefly, 5 × 10 3 HuH-7 cells/well were plated in 16-well E-Plate and placed onto the multi-plate Real-Time Cell Analyzer located inside a tissue culture incubator.…”

Endogenous Heparinoids Detected by anti-Xa Activity are Present in Blood during Acute Variceal Bleeding in Cirrhosis. A Prospective Study

“…We did not nd any correlation between raised anti-Xa levels and tumor size, or number of nodes, but this could be a type 2 error. Experimental studies on hepatoma cells have shown that heparin inhibits cellular invasion [28], whereas clinical studies have described a positive e ect of LMWHs on cancer survival [29]. Considering that raised anti Xa levels were predictive for 6-week mortality and that approximately 57% of variceal bleeders with raised anti Xa levels had HCC, this might re ect the con rmed strong association between HCC and bleeding-related mortality in portal hypertensive bleeding [30,31].…”