epidermal-growth-factor and Esophageal-Squamous-Cell-Carcinoma

Homeobox A7 (HOXA7) plays essential roles in multiple malignancies and was reported to be overexpressed in esophageal squamous cell carcinoma (ESCC). However, its functions in the ESCC tumor microenvironment remain to be explored. In this study, we showed that HOXA7 was overexpressed in ESCC among HOXA family members and correlated with tumor-associated macrophage (TAM) infiltration both in The Cancer Genome Atlas database and ESCC clinical samples. Moreover, transactivation of C-C motif chemokine ligand 2 (CCL2) by HOXA7 was identified (real-time quantitative PCR [RT-qPCR], western blot analysis, ELISA, and ChIP-qPCR), which was detected to drive chemotaxis and M2 polarization of macrophages both in vitro (Transwell assay) and in vivo (xenograft tumors models). In addition, CCL2 triggers macrophage expression of epidermal growth factor (EGF) (RT-qPCR and ELISA), which promotes tumor proliferation and metastasis by activating its receptor EGFR. In addition, EGF-induced ESCC cell proliferation and migration can be abrogated by HOXA7 knockdown (CCK-8 proliferation assay, EdU fluorescence, and Transwell assay). These results indicate a novel mechanistic role of HOXA7 in the cross-talk between ESCC and TAMs, which could be an underlying therapeutic target for ESCC.

Topics: Cell Line, Tumor; Cell Movement; Cell Proliferation; Epidermal Growth Factor; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Gene Expression Regulation, Neoplastic; Genes, Homeobox; Humans; Ligands; Transcription Factors; Tumor Microenvironment; Tumor-Associated Macrophages

Esophageal squamous cell carcinoma (ESCC) presents with lymph node metastasis in the early stages, limiting the opportunities for curative local resection, including endoscopic submucosal dissection (ESD). ESD is regarded as the standard treatment for early-stage ESCCs. However, radical surgery is recommended when lymph node metastasis risk exists. More efforts are needed to find the markers for early prediction and clarify the molecular mechanism underlying the pathogenesis of lymph node metastasis. Recently, aberrant regulation of gene expression by histone methylation modifiers has emerged as an important mechanism for cancer metastasis. Herein, we demonstrated that mixed-lineage leukemia 2 (MLL2) positively regulates gene expression programs associated with ESCC cell migration. MLL2 interacts with RelA in the nucleus to enhance transcription of stanniocalcin-1 (STC1) and to facilitate cancer metastasis. Meanwhile, MLL2 knockdown resulted in a significant decrease in the migration of ESCC cells. Clinically, high level of MLL2 was significantly associated with early-stage ESCC lymph node metastasis. In summary, these findings discovered a previously unidentified molecular pathway underlying the coordinated regulation of metastasis-related STC-1 expression by MLL2 and RelA and highlighted the critical role of MLL2 in ESCC.

Topics: Animals; Carcinogenesis; Cell Line, Tumor; Cell Movement; DNA-Binding Proteins; Epidermal Growth Factor; Esophageal Squamous Cell Carcinoma; Extracellular Signal-Regulated MAP Kinases; Gene Silencing; Glycoproteins; Humans; Lymphatic Metastasis; Male; Mice, Nude; Neoplasm Invasiveness; Neoplasm Proteins; Promoter Regions, Genetic; Protein Binding; Transcription Factor RelA; Transcription, Genetic; Tumor Stem Cell Assay

Lymphatic metastasis is an important determinant of aggressive malignant tumors. Identification of key genes that regulate carcinoma cell metastasis will aid in understanding progression of esophageal squamous cell carcinoma (ESCC). Guanylate-binding protein 1 (GBP1) is a GTP-binding protein family member with high GTPase activity. While the role of GBP1 has been demonstrated in colorectal cancer (CRC) and ovarian cancer, such a role has not been identified in ESCC. Herein, we assessed GBP1 expression in ESCC via immunohistochemistry (IHC) analysis of 215 clinical ESCC specimens and found that the expression of GBP1 was significantly upregulated in lymph node metastatic ESCCs compared with non-metastatic cases. We further demonstrated that GBP1 expression was increased with epidermal growth factor (EGF) stimulus in ESCC cell lines and had a positive correlation with EGFR expression in ESCC tissue samples. Finally, we found that overexpression of GBP1 in ESCC cells induced more lymph node metastasis in an animal model. In summary, our results demonstrate that GBP1 promotes lymph node metastasis and has a positive correlation with EGFR expression in ESCC.

Topics: Animals; Carcinoma, Squamous Cell; Cell Line, Tumor; Disease Models, Animal; Epidermal Growth Factor; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Female; Gene Expression Regulation, Neoplastic; GTP-Binding Proteins; Humans; Lymph Nodes; Lymphatic Metastasis; Male; Mice, Inbred BALB C; Mice, Nude; Neovascularization, Pathologic; RNA, Messenger; Up-Regulation

To investigate the association between genetic polymorphisms of growth factor-related genes and prognosis in patients with advanced esophageal squamous cell carcinoma (ESCC).. A total of 334 ESCC patients with advanced tumor stages (stages IIB, III and IV) were enrolled in the study. The genotypes of 14 candidate single nucleotide polymorphisms (SNPs) involved in growth factor-related functions were analyzed using iPLEX Gold technology from the genomic DNA of peripheral leukocytes, and were correlated with the clinical outcome of patients. Serum levels of growth factors were examined by enzyme-linked immunosorbent assay (ELISA).. The genetic polymorphisms of EGF:rs4444903, EGF:rs2237051 and VEGF:rs2010963 showed significant associations with overall survival (OS) of advanced ESCC patients (A/A+ A/G vs. GG, [HR = 0.77, 95% CI = 0.60-0.99, P = 0.039 for rs4444903; A/G+ G/G vs. A/A, [HR = 0.74, 95% CI = 0.58-0.95, P = 0.019 for rs2237051; G/G+G/C vs. C/C, [HR] inves = 0.69, 95% CI = 0.50-0.95, P = 0.023 for rs2010963). EGFR:rs2227983 and 3 SNPs of PIK3CA also showed borderline significant correlation with OS of advanced ESCC patients (P = 0.058 for rs2227983; P = 0.069, 0.091 and 0.067 for rs6443624, rs7651265 and rs7621329 of PIK3CA respectively). According to cumulative effect analysis of multiple SNPs, patients carrying 4 unfavorable genotypes exhibited more than a 3-fold increased risk of mortality. Finally, both EGF and VEGF expression levels significantly associated with patient mortality.. The genetic variants and expression levels of EGF and VEGF can serve as prognostic predictors in patients with advanced ESCC, and thus provide more information for optimizing personalized therapies for patients with ESCC.

Topics: Aged; Carcinoma, Squamous Cell; Disease-Free Survival; Epidermal Growth Factor; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Female; Genetic Predisposition to Disease; Humans; Kaplan-Meier Estimate; Male; Middle Aged; Multivariate Analysis; Neoplasm Recurrence, Local; Neoplasm Staging; Polymorphism, Single Nucleotide; Prognosis; Vascular Endothelial Growth Factor A

Epithelial-mesenchymal transition (EMT) is a developmental program that is associated with esophageal squamous cell carcinoma (ESCC) progression and metastasis. Recently, C/EBPβ has been reported to be an EMT inducer in cancer. However, the detailed molecular mechanisms remain unclear. Here, we report for the first time, that the truncated CCAAT-enhancer-binding protein β (C/EBPβ) LIP isoform is abnormally overexpressed and correlated with cancer metastasis in clinical specimens of human ESCC. Furthermore, we demonstrate that C/EBPβ LIP mediates epithelial growth factor (EGF)-induced EMT and increases migration and invasion of esophageal cancer cells in a manner that is dependent on miR-203 inactivation. Finally, we identified miR-203 as a direct target of C/EBPβ LIP. Disruption of C/EBPβ LIP attenuated the EGF-mediated decrease in miR-203, whereas overexpression of C/EBPβ LIP alone markedly suppressed miR-203. In addition, we demonstrated that C/EBPβ LIP inhibited miR-203 transcription by directly interacting with a conserved distal regulatory element upstream of the miR-203 locus, and in doing so, orchestrated chromatin remodeling. In conclusion, our results have revealed a new regulatory mechanism that involves C/EBPβ-LIP-mediated downregulation of miR-203, which plays a key role in EMT and metastasis.

Topics: Carcinoma, Squamous Cell; CCAAT-Enhancer-Binding Protein-beta; Cell Line, Tumor; Epidermal Growth Factor; Epithelial Cells; Epithelial-Mesenchymal Transition; Esophageal Neoplasms; Esophageal Squamous Cell Carcinoma; Gene Expression Regulation, Neoplastic; Humans; MicroRNAs