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
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
复制标题
表皮生长因子和肿瘤坏死因子α通过NF-kappa B/p65协同诱导纤连蛋白协同促进肝癌细胞系的运动
DOI：
10.1016/j.bbagen.2017.08.010
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发表时间：
2017-11-01
影响因子：
3
通讯作者：
Du, Jun
中科院分区：
文献类型：
--
作者：
Liu, Zong-Cai;Ning, Fen;Du, Jun
背景：肝细胞癌（HCC）细胞与其微环境之间的相互作用在肿瘤转移中起着重要作用。肝癌微环境中含有丰富的表皮生长因子（EGF）和肿瘤坏死因子α（TNF α），EGF和TNF α可能协同作用而非单独作用，影响肿瘤细胞的生物学行为。采用Western blotting、免疫荧光、qRT-PCR、创伤愈合划痕和侵袭试验以及染色质免疫沉淀试验研究EGF和TNF α在HCC细胞体外运动中的联合作用。在体外，EGF和TNF α主要通过协同诱导细胞外基质糖蛋白纤连蛋白（FN）协同促进HCC细胞的运动。EGF和TNF α共同促进了NF-κ B/p65的核转位和PKC介导的磷酸化，使NF-κ B/p65与FN启动子的-356 bp ~-259 bp片段结合，从而导致HCC细胞中FN启动子的活性显著增加。EGF和TNF α主要通过NF-κ B B/p65介导的协同诱导FN而协同促进肝癌细胞的运动。
Background: 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 alpha (TNF alpha), which may cooperatively, rather than individually, interact with tumor cells to influence their biological behavior.Methods: Immunohistochemistry was performed to study the expression of EGF and TNF alpha 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 TNF alpha in the motility of HCC cells in vitro.Results: We demonstrated that both EGF and TNF alpha were highly expressed in HCCs, and HCCs with higher expression of both EGF and TNFa were more frequently rated as high-grade tumors. In vitro, EGF and TNF alpha cooperatively promoted the motility of HCC cells mainly via synergistic induction of an extracellular matrix glycoprotein fibronectin (FN). Mechanistically, EGF and TNF alpha jointly increased the nuclear translocation and PKC mediated phosphorylation of NF-kappa B/p65 which could bind to the -356 bp to -259 bp fragment of the FN promoter, leading to a markedly increased activity of the FN promoter in HCC cells.Conclusions: HCCs with higher expression of both EGF and TNF alpha were more frequently rated as high-grade tumors. EGF and TNF alpha cooperatively promoted the motility of HCC cells mainly through NF-kappa B/p65 mediated synergistic induction of FN in vitro.General Significance: These findings highlight the crosstalk between EGF and TNF alpha in promoting HCC, and provide potential targets for HCC prevention and treatment.