It has been proven that CC chemokine receptor 7 (CCR7) is closely related with the lymph node metastasis of non-small cell lung cancer (NSCLC), but the mechanism of NSCLC metastasis is not very clear. The aim of this study is to investigate the expressions of CCR7 and MMP-9 in NSCLC and the relationship of their expressions, and to explore the mechanism of CCR7 promoting NSCLC metastasis.
The expressions of CCR7 and MMP-9 protein were detected in 90 specimens of human primary NSCLC by immunohistochemical SP method. Human large lung cell line BE1 cells were pre-incubated with CCL19 for 24 h; the changes of MMP-9 were detected by RT-PCR and Western blot.
Immunohistochemistry showed that CCR7 was distributed in cytoplasm and/or membrane of tumor cells and MMP-9 was distributed in cytoplasm of cancer cells. The expressions of CCR7 and MMP-9 protein were found to be 70% (63/90) and 65.5% (59/90) in NSCLC, respectively. The expressions of CCR7 and MMP-9 protein were closely related to the clinical stages (P=0.003, P=0.001) and lymph node metastasis (P=0.004, P=0.003) of NSCLC, but there was no correlation with age, gender, histology (P > 0.05). Furthermore, a significant correlation was found between CCR7 and MMP-9 (r=0.342, P=0.001). In addition, the expressions of MMP-9 mRNA and protein levels were increased in CCL19 pre-incubated group (P < 0.05).
The expressions of CCR7 and MMP-9 are significantly associated with NSCLC invasion and metastasis. The upregulation of MMP-9 is regulated by CCR7 in NSCLC.
[Show abstract][Hide abstract] ABSTRACT: Although ZNF746, also known as Parkin-interacting substrate (PARIS), has been reported to suppress peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) and its target gene NRF-1 leading to the neurodegeneration in Parkinson's disease, its function in tumorigenesis has yet to be investigated. Thus, in the present study, the role of ZNF746 in the invasion and epithelial to mesenchymal transition (EMT) in H460 non-small cell lung cancer (NSCLC) cells was investigated. Invasion assay showed that inhibition of ZNF746 using siRNA transfection inhibited the invasion of H460 NSCLC cells using Boyden chamber. Quantitative PCR (qPCR) analysis revealed that the silencing of ZNF746 attenuated the expression of matrix metalloproteinase (MMP)1, MMP2 and MMP9, but not MMP7, in H460 NSCLC cells. Immunoblotting assay revealed that the expression of E-cadherin and β-catenin of epithelial phenotype was upregulated, while Slug was downregulated in ZNF746 siRNA-transfected H460 NSCLC cells. Accordingly, the mRNA expression of E-cadherin was upregulated while vimentin or Slug, Twist, ZEB as EMT key transcriptional factors were suppressed in ZNF746 siRNA-transfected H460 NSCLC cells. Also, mRNA expression of transcriptional marker Nanog and Octamer-binding transcription factor 4 (OCT4), known to enhance malignancy and metastasis in lung adenocarcinoma, was suppressed in ZNF746 siRNA-transfected H460 NSCLC cells. Notably, the endogenous expression of ZNF746 was induced in parallel with Twist at the protein level during hypoxia. Overall, our findings suggest that inhibition of ZNF746 suppresses the invasion and EMT molecules in H460 NSCLC cells and ZNF746 may be an important target molecule in lung tumorigenesis.
[Show abstract][Hide abstract] ABSTRACT: The mechanisms leading to squamous cell carcinoma of head and neck (SCCHN) metastasis are not fully understood. Although evidence shows that the chemokine receptor 7 (CCR7) and its ligand CCL19 may regulate tumor dissemination, their role is not clearly defined in SCCHN. Matrix metalloproteinases break consisting of tissue barrier to the surrounding tissue invasion and metastasis by destroying the balance of matrix degradation of the basement membrane of tumor cells and extracellular matrix (ECM). We used chemotaxis and migration assays, western blotting, gelatin zymography, actin polymerization assay, immunofluorescence staining and immunohistochemical analysis to explore whether MMP-9 can be activated by CCL19 (CCR7's ligand) and its role in SCCHN. The experiments were performed in the metastatic SCCHN cell line PCI-37B after pre-incubation of the cells with CCL19 and SB-3CT (inhibitor of MMP-9). Our results demonstrated that CCR7 favors PCI-37B cell chemotaxis and migration, upregulation of MMP-9 protein and motivates the activity of MMP-9 protein, induces reorganization of the actin cytoskeleton and upregulation of MMP-9 protein. SB-3CT can block all these effects. Collectively, our data indicated that CCR7 regulates cell chemotaxis and migration via MMP-9 in metastatic SCCHN, and these results provide a basis for new strategies in preventing metastases of SCCHN.
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