N-terminal and C-terminal heparin-binding domain polypeptides derived from fibronectin reduce adhesion and invasion of liver cancer cells

• Published in: BMC cancer Vol. 10; no. 1; p. 552
• Main Authors: Tang, Nan-Hong, Chen, Yan-Lin, Wang, Xiao-Qian, Li, Xiu-Jin, Wu, Yong, Zou, Qi-Lian, Chen, Yuan-Zhong
• Format: Journal Article
• Language: English
• Published: England BioMed Central Ltd 13.10.2010; BioMed Central; BMC
• Subjects: Animals; Cancer; Carcinoma, Hepatocellular - metabolism; Cell Adhesion; Cell Line, Tumor; Cloning; Collagen - chemistry; Development and progression; Drug Combinations; Fibronectins; Fibronectins - chemistry; Genetic aspects; Glycoproteins - chemistry; Heparin - chemistry; Humans; Laminin - chemistry; Liver cancer; Liver Neoplasms - metabolism; Liver Neoplasms - pathology; Male; Metastasis; Mice; Mice, Nude; Neoplasm Invasiveness; Neoplasm Metastasis; Peptides - chemistry; Physiological aspects; Protein Structure, Tertiary; Proteins; Proteoglycans - chemistry
• ISSN: 1471-2407; 1471-2407
• DOI: 10.1186/1471-2407-10-552

Fibronectin (FN) is known to be a large multifunction glycoprotein with binding sites for many substances, including N-terminal and C-terminal heparin-binding domains. We investigated the effects of highly purified rhFNHN29 and rhFNHC36 polypeptides originally cloned from the two heparin-binding domains on the adhesion and invasion of highly metastatic human hepatocellular carcinoma cells (MHCC97H) and analyzed the underlying mechanism involved. The MHCC97H cells that adhered to FN in the presence of various concentrations of rhFNHN29 and rhFNHC36 polypeptides were stained with crystal violet and measured, and the effects of rhFNHN29 and rhFNHC36 on the invasion of the MHCC97H cells were then detected using the Matrigel invasion assay as well as a lung-metastasis mouse model. The expression level of integrins and focal adhesion kinase (FAK) phosphotyrosyl protein was examined by Western blot, and the activity of matrix metalloproteinases (MMPs) and activator protein 1 (AP-1) was analyzed by gelatin zymography and the electrophoretic mobility band-shift assay (EMSA), respectively. Both of the polypeptides rhFNHN29 and rhFNHC36 inhibited adhesion and invasion of MHCC97H cells; however, rhFNHC36 exhibited inhibition at a lower dose than rhFNHN29. These inhibitory effects were mediated by integrin αvβ3 and reversed by a protein tyrosine phosphatase inhibitor. Polypeptides rhFNHN29 and rhFNHC36 abrogated the tyrosine phosphorylation of focal adhesion kinase (p-FAK) and activation of activator protein 1 (AP-1), resulting in the decrease of integrin αv, β3 and β1 expression as well as the reduction of MMP-9 activity. Polypeptides rhFNHN29 and rhFNHC36 could potentially be applicable to human liver cancer as anti-adhesive and anti-invasive agents.