CCR7 mediates the TNF-α-induced lymphatic metastasis of gallbladder cancer through the "ERK1/2 - AP-1" and "JNK - AP-1" pathways.
CCR7 mediates the TNF-α-induced lymphatic metastasis of gallbladder cancer through the "ERK1/2 - AP-1" and "JNK - AP-1" pathways.
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DOI：
10.1186/s13046-016-0318-y
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发表时间：
2016-03-24
期刊：
影响因子：
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通讯作者：
Chen Y
中科院分区：
文献类型：
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作者：
Hong H;He C;Zhu S;Zhang Y;Wang X;She F;Chen Y
CC趋化因子受体7（CCR 7）在多种肿瘤中高度表达，并与淋巴结转移呈正相关，在细胞定向运动中起重要作用。炎症细胞因子肿瘤坏死因子（TNF）-α促进胆囊癌（GBC）的肿瘤进展和淋巴结转移。但CCR 7在GBC中的表达尚不清楚，其在TNF-α诱导的GBC淋巴道转移中的作用有待进一步研究。采用免疫组化法检测临床标本中CCR 7的表达，分析CCR 7与临床病理因素及胆汁中TNF-α水平的关系。用不同浓度的TNF-α处理后，通过蛋白质印迹法测定GBC细胞系中CCR 7的表达。采用相对荧光素酶报告基因分析、定点突变和染色质免疫沉淀等方法分析CCR 7的启动子活性和转录调控。MAPKs抑制剂用于探索AP-1的上游信号分子。我们建立了稳定表达慢病毒CCR 7 shRNA的NOZ细胞系，并通过transwell实验和动物实验研究TNF-α -CCR 7轴在GBC细胞向淋巴系统迁移中的作用。CCR 7在胆囊癌组织中高表达。CCR 7的高表达与美国癌症联合委员会（AJCC）分期和淋巴结转移有关。此外，我们发现胆囊癌组织中CCR 7的表达与胆汁中TNF-α的水平呈正相关，TNF-α通过“ERK 1/2-AP-1”和“JNK-AP-1”途径增强CCR 7的启动子活性和蛋白表达。最后，我们揭示了TNF-α可以通过上调CCR 7在体外和体内促进GBC细胞向淋巴管内皮细胞或淋巴结迁移。我们的研究表明，CCR 7在GBC中高表达，并通过“TNF-α -ERK 1/2 - AP-1 -CCR 7”和“TNF-α -JNK-AP-1 -CCR 7”途径介导TNF-α诱导的GBC淋巴道转移。
CC-chemokine receptor 7 (CCR7), which plays an important role in cell directional movement, is highly expressed in various cancers and positively related to lymph node metastasis. The inflammatory cytokine tumour necrosis factor (TNF)-α promotes tumour progression and lymph node metastasis in gallbladder cancer (GBC). However, the expression of CCR7 in GBC is unclear, and its role in the TNF-α-induced lymphatic metastasis of GBC requires further research. The expression of CCR7 in clinical samples was detected by immunohistochemistry, and the relationship between CCR7 and clinicopathological factors or the TNF-α level of the bile was analyzed. After treatment with various concentrations of TNF-α, CCR7 expression in GBC cell lines was measured by Western blotting. The relative luciferase reporter assay, site-directed mutagenesis and chromatin immunoprecipitation were used to analyze the promoter activity and transcriptional regulation of CCR7. MAPKs inhibitors were used to explore the upstream signalling molecules of AP-1. We established a NOZ cell line stably expressing lentiviral CCR7 shRNA that effectively silenced the expression of CCR7, and to determine the role of TNF-α - CCR7 axis in the migration of GBC cells to the lymphatic system by transwell assays and animal experiments. CCR7 was highly expressed in GBC samples. Higher expression of CCR7 was associated with American Joint Committee on Cancer (AJCC) staging and lymph node metastasis. Moreover, we found that CCR7 expression in GBC tissue was positively correlated with the levels of TNF-α in the bile, and that TNF-α enhanced the promoter activity and protein expression of CCR7 through the “ERK1/2-AP-1” and “JNK-AP-1” pathways. Finally, we revealed that TNF-α could promote GBC cell migration to lymphatic endothelial cells or lymph nodes through upregulation of CCR7 in vitro and in vivo. Our study suggests that CCR7 is highly expressed in GBC, and mediates the TNF-α-induced lymphatic metastasis of GBC through the “TNF-α - ERK1/2 - AP-1 - CCR7” and “TNF-α - JNK - AP-1 - CCR7” pathways.