WSS25 Inhibits Growth of Xenografted Hepatocellular Cancer Cells in Nude Mice by Disrupting Angiogenesis via Blocking Bone Morphogenetic Protein (BMP)/Smad/Id1 Signaling

Abstract

Hepatocellular cancer (HCC)3 is the fourth leading cause of death from cancer worldwide and the second most lethal malignant cancer in China since the 1990s (1). However, there are few treatment options aside from surgery (1). HCC is characterized by hypervascularity, which can distinguish HCC from benign lesions by angiography. The strategy of blocking angiogenesis in HCC to inhibit tumor growth has been used for more than 20 years in the clinic (2). However, there are no ideal antiangiogenesis chemotherapeutic agents developed thus far for HCC therapy.Id1 is one of the inhibitors of DNA binding proteins (Ids), which belongs to the basic helix loop helix transcriptional factor superfamily (3). There are four members, Id1, Id2, Id3, and Id4, in the Ids family. Accumulating evidence show that Id1 is overexpressed in solid tumors and their supporting vasculatures (4, 5). Although essential during embryo development, Id1 expression is extremely low in adult tissues, including the quiescent endothelial cells (6). Partial loss of Id1 by genetic manipulation in mice effectively inhibits tumor angiogenesis (6). Id1 ablation leads to the impairment of bone marrow endothelial progenitor cell recruitment and mobilization to form tumor vasculature (7,8). Id1 also has been implicated in the regulation of cell senescence, drug resistance, tumor cell invasion, and apoptosis resistance (9). Interestingly, Id1 has been demonstrated to be highly expressed in malignant HCC and to promote angiogenesis in HCC (10, 11). Thus, Id1 is a rational target for the development of antiangiogenesis therapeutics for HCC.Heparan sulfate proteoglycans are glycoconjugates composed of protein cores to which heparan sulfate (HS) chains are attached. Direct genetic evidence supports that heparan sulfate proteoglycans are necessary for tumor angiogenesis (12). Heparan sulfate proteoglycans, especially those located on the cell surface, act as co-receptors through their HS chains (13). HS chains are negatively charged polymers composed of hexaronic acid and glucosamine disaccharide repeats. Due to their negative charge, they bind multiple functional proteins, including Tables 1 and 2

Lignans from Schisandra sphenathera Rehd. et Wils. and semisynthetic schisantherin A analogues: Absolute configuration, and their estrogenic and anti-proliferative activity

“…The effect on cell proliferation was determined using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] method [44].…”

Quantitative Phosphoproteomics Analysis Reveals Broad Regulatory Role of Heparan Sulfate on Endothelial Signaling

“…Wound Healing Recovery Assay (28,29)-MLECs (2 ϫ 10 5 cells per well) were seeded into a six-well plate and reached confluency after culture for 24 h. A wound gap was made by scratching with a T-200 tip in each well. After removing debris by PBS washing, the cells were cultured in DMEM medium supplemented with 10% FBS.…”

“…Cell Proliferation Assay (28,29)-Ext1f/f or Ext1Ϫ/Ϫ MLECs (3000 cells/well) were seeded into a 96-well plate. Cell growth was assessed by colorimetric assay using the Cell Counting Kit-8 kit (Dojindo Molecular Technologies, Santa Clara, CA).…”

“…Tube Formation (28,29)-MLECs (3 ϫ 10 4 /well) in DMEM medium containing 5% FBS were seeded on 96-well plate coated with growth factor reduced matrigel. After 6 h, cell morphology in culture was photographed under the Nikon Eclipse TE2000-S inverted microscope.…”

WSS25, a sulfated polysaccharide, inhibits RANKL-induced mouse osteoclast formation by blocking SMAD/ID1 signaling

“…A previous study in our lab demonstrated that WSS25 binds to BMP-2 and that its receptors inhibited xenografted hepatocellular cancer cell growth and disrupted angiogenesis [26][27][28] . As we mentioned above, BMP-2 is also a crucial factor in bone remodeling.…”