Epidermal growth factor and tumor necrosis factor α cooperatively promote the motility of hepatocellular carcinoma cell lines via synergistic induction of fibronectin by NF-κB/p65

• Published in: Biochimica et biophysica acta. General subjects Vol. 1861; no. 11; pp. 2568 - 2582
• Main Authors: Liu, Zong-Cai, Ning, Fen, Wang, Hai-Fang, Chen, Dan-Yang, Cai, Yan-Na, Sheng, Hui-Ying, Lash, Gendie E., Liu, Li, Du, Jun
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.11.2017
• Subjects: Carcinoma, Hepatocellular - genetics; Carcinoma, Hepatocellular - pathology; Cell Line, Tumor; cell lines; Cell Movement - genetics; chromatin; epidermal growth factor; Epidermal growth factor (EGF); Epidermal Growth Factor - genetics; extracellular matrix; Fibronectin; fibronectins; Fibronectins - biosynthesis; Fibronectins - genetics; fluorescent antibody technique; Gene Expression Regulation, Neoplastic; glycoproteins; Hepatocellular carcinoma (HCC); hepatoma; Humans; Liver Neoplasms - genetics; Liver Neoplasms - pathology; Metastasis; neoplasm cells; NF-kappa B - genetics; Phosphorylation; precipitin tests; protein kinase C; quantitative polymerase chain reaction; reverse transcriptase polymerase chain reaction; Transcription Factor RelA - genetics; Tumor microenvironment; Tumor necrosis factor α (TNFα); tumor necrosis factor-alpha; Tumor Necrosis Factor-alpha - genetics; Western blotting; SB2; Tumor microenvironment; N-cad; Hepatocellular carcinoma (HCC); Epidermal growth factor (EGF); FN; Metastasis; EGF/TNFα; LY; Fibronectin; E+T; PD; BAY; Vim; Tumor necrosis factor α (TNFα); E-cad; GF
• ISSN: 0304-4165; 1872-8006
• DOI: 10.1016/j.bbagen.2017.08.010

The interaction between hepatocellular carcinoma (HCC) cells and their microenvironment plays a fundamental role in tumor metastasis. The HCC microenvironment is rich in epidermal growth factor (EGF) and tumor necrosis factor α (TNFα), which may cooperatively, rather than individually, interact with tumor cells to influence their biological behavior. Immunohistochemistry was performed to study the expression of EGF and TNFα in HCCs. Western blotting, immunofluorescence, qRT-PCR, wound healing scratch and invasion assay, and chromatin immunoprecipitation assays were used to study the combined roles of EGF and TNFα in the motility of HCC cells in vitro. We demonstrated that both EGF and TNFα were highly expressed in HCCs, and HCCs with higher expression of both EGF and TNFα were more frequently rated as high-grade tumors. In vitro, EGF and TNFα cooperatively promoted the motility of HCC cells mainly via synergistic induction of an extracellular matrix glycoprotein fibronectin (FN). Mechanistically, EGF and TNFα jointly increased the nuclear translocation and PKC mediated phosphorylation of NF-κB/p65 which could bind to the −356bp to −259bp fragment of the FN promoter, leading to a markedly increased activity of the FN promoter in HCC cells. HCCs with higher expression of both EGF and TNFα were more frequently rated as high-grade tumors. EGF and TNFα cooperatively promoted the motility of HCC cells mainly through NF-κB/p65 mediated synergistic induction of FN in vitro. General Significance: These findings highlight the crosstalk between EGF and TNFα in promoting HCC, and provide potential targets for HCC prevention and treatment. •Both EGF and TNFα are highly expressed in hepatocellular carcinoma (HCC).•HCCs with higher level of both EGF and TNFα are more rated as high-grade tumors.•EGF and TNFα jointly promote the motility of HCC cells via fibronectin (FN).•p65 is crucial for synergistic induction of FN by EGF and TNFα.•EGF and TNFα jointly increase the nuclear localization and phosphorylation of p65.