Weidong Shen

VEGF-C promotes the development of esophageal cancer via regulating CNTN-1 expression.

Vascular endothelial growth factor C (VEGF-C) is a key regulator of angiogenesis and lymphangiogenesis. VEGF-C is also implicated in the development of esophageal cancer. We investigated the mRNA levels of VEGF-C and its receptors in 38 esophageal squamous cell carcinoma specimens (ESCCs) and matched adjacent normal esophageal tissues via real-time PCR. The mRNA levels of VEGF-C, VEGFR-2 and VEGFR-3 were significantly upregulated in ESCCs versus respective side normal tissues. To explore the influence of VEGF-C on esophageal cancer progression, the expression of VEGF-C was manipulated in esophageal cancer cell lines TE-1 and Eca-109. VEGF-C transcription, translation and secretion were significantly enhanced in cells stably transfected with a VEGF-C overexpression vector or attenuated in VEGF-C shRNA-transfected cell lines. In vitro, TE-1 cells stably transfected with a VEGF-C overexpression vector exhibited an increased rate of cell proliferation, migration and focus formation, whereas knockdown of VEGF-C inhibited cell proliferation, migration and focus formation. Similar results were obtained for Eca-109 cells. VEGF-C mediated biological function through transcription of CNTN-1, which is implicated in tumor invasion and metastasis. The expression of VEGF-C was correlated with that of CNTN-1 and cell proliferation and migration induced by VEGF-C were reversed by silencing of CNTN-1. In addition, nude mice inoculated with VEGF-C shRNA-transfected cells exhibited a significantly decreased tumor size in vivo via reduced VEGFR-2 and VEGFR-3 phosphorylation and microvessel formation. VEGF-C upregulation may be involved in esophageal tumor progression. Vector-based RNA interference (RNAi) targeting VEGF-C is a potential therapeutic method for human esophageal carcinoma.

Expression of VEGF-C Correlates with a Poor Prognosis Based on Analysis of Prognostic Factors in 73 Patients with Esophageal Squamous Cell Carcinomas

OBJECTIVE The purpose of this study was to specifically investigate the clinicopathological role of expression of vascular endothelial growth factor-C (VEGF-C) as well as the correlation with clinical outcomes in esophageal squamous cell carcinomas (ESCCs). METHODS Seventy-three patients with ESCC resected in our institute were included in this study. Formalin-fixed paraffin-embedded specimens were stained for VEGF-C and the correlation between the staining, its clinicopathological parameters and its prognostic power were analyzed statistically. RESULTS Of the 73 ESCC patients studied, 39 cases (53.4%) were strongly positive for VEGF-C. Six cases (8.2%) were negative and 28 cases (38.4%) revealed unclear weak reactions. All 34 cases were included in the negative group (46.6%). VEGF-C expression correlated with histological grade (P = 0.005), depth of tumor invasion (pT) (P = 0.021), lymph node metastasis (pN) (P = 0.002) and lymphatic invasion (P = 0.008). The median overall survival of 39 patients who had positive staining for tumor cell VEGF-C and 34 patients who had negative staining were 10.4 months (95% CI, 6.9-13.9 months) and 28.5 months (95% CI, 12.6-44.4 months), respectively (P = 0.003). In univariate analysis by log-rank test, histological grade, pN, stage, lymphatic invasion and VEGF-C were significant prognostic factors (P = 0.047, 0.007, 0.018, 0.002 and 0.003, respectively.). In multivariate analysis, high VEGF-C expression (P = 0.0451) maintained its independent prognostic influence on overall survival, as well as pN status (P = 0.0029). CONCLUSIONS Expression of VEGF-C is related to histological grade, pT, pN and lymphatic invasion, and is a prognostic indicator for ESCC.

Correlation of E-cadherin and CD44v6 expression with clinical pathology in esophageal carcinoma.

Cell adhesion, important for maintaining tissue architecture, plays a role in numerous cancers and particularly in tumor progression. In the present study, we investigated perturbations in the expression of two important adhesion proteins, epithelial (E)-cadherin and CD44v6, in esophageal carcinoma (EC). EC specimens were obtained from 42 patients undergoing resection of EC; both cancer and adjacent normal tissue were collected. Expression of E-cadherin and CD44v6 was detected by immunohistochemistry and the correlation between the expression of these two proteins and their individual relationships with pathology were determined. E-cadherin expression in EC tissue was significantly less common than in adjacent normal tissue. Furthermore, absence of E-cadherin expression was correlated with infiltration depth, lymph node metastasis, distant metastases and TNM stage (P<0.05), but not with gender, age, differentiation or tumor size. By contrast, CD44v6 expression in EC was significantly higher than that in adjacent normal tissue and was correlated with differentiation, distant metastases and TNM stage (P<0.05), but not with other clinicopathological parameters. Additionally, we observed a negative correlation between E-cadherin and CD44v6 expression in EC (P<0.05). Based on their correlations with pathology, we suggest that the expression of E-cadherin and CD44v6 is important roles in promoting the infiltration and metastasis of EC.

Contactin-1 (CNTN-1) Overexpression is Correlated with Advanced Clinical Stage and Lymph Node Metastasis in Oesophageal Squamous Cell Carcinomas

OBJECTIVE Oesophageal squamous cell carcinoma is one of the deadliest malignancies worldwide. Contactin-1, a neural adhesion molecule, is implicated in tumour invasion and metastasis. The purpose of this study was to investigate the expression of CNTN-1 in normal and cancerous oesophageal tissue, and the potential relevance to clinicopathological features. METHODS Thirty normal oesophageal tissue samples and 82 primary oesophageal squamous cell carcinoma tissue samples were included in this study. The expression levels of CNTN-1, VEGF-C and HIF-1α messenger RNA were determined using reverse transcriptase-polymerase chain reaction and quantitative real-time polymerase chain reaction. The expression of the CNTN-1 protein was measured using immunohistochemistry. RESULTS The expression of CNTN-1 messenger RNA was significantly increased in the tumour tissue compared with the normal oesophageal tissue (P=0.001). The oesophageal squamous cell carcinoma tissue consistently showed higher CNTN-1 protein levels. The CNTN-1 expression correlated with the oesophageal squamous cell carcinoma stage (P=0.006), lymph node metastasis (P=0.018) and lymphatic invasion (P=0.035). The messenger RNA level of CNTN-1 correlated significantly with those of VEGF-C and HIF-1α. CONCLUSIONS The expression of CNTN-1 is upregulated in the oesophageal squamous cell carcinoma tissue and related to stage, lymph node metastasis and lymphatic invasion. Thus, CNTN-1 may be involved in the progression and pathogenesis of oesophageal squamous cell carcinoma.

Upregulation of PAX2 promotes the metastasis of esophageal cancer through interleukin-5.

Background/Aims: This study investigated the clinical relevance and biological function of paired box gene 2 (PAX2) in esophageal squamous cell carcinoma (ESCC). Methods/Results: Results showed that PAX2 expression was significantly increased in tumor tissues and that its expression correlated with the ESCC stage (P = 0.001), lymph node metastasis (pN, P = 0.019) and lymphatic invasion (P = 0.005) in 120 ESCC tissue specimens by immunohistochemistry. Furthermore, PAX2 overexpression resulted in markedly reduced cell proliferation but increased metastasis capacity in ESCC TE-1 and Eca-109 cells. Knockdown of PAX2 expression with a short hairpin RNA confirmed a role in the promotion of metastasis in ESCC cells. mRNA microarray screening revealed that PAX2 overexpression affected multiple genes that function in multiple pathways. Interleukin-5 (IL-5), which was induced by PAX2 and has been shown to promote tumor metastasis, was further studied in greater detail. Two PAX2 binding sites were identified in the IL-5 promoter, and PAX2 was observed to stimulate IL-5 promoter activity and IL-5 expression in esophageal cancer cells. Chromatin immunoprecipitation (ChIP) confirmed the direct binding of PAX2 in the IL-5 promoter. The expression of PAX2 mRNA significantly correlated with that of IL-5 in normal esophageal and ESCC tissues. Conclusion: These findings demonstrate that PAX2 is overexpressed in esophageal carcinoma and IL-5 is identified as PAX2s effector for metastasis.

Effects of diphyllin as a novel V-ATPase inhibitor on TE-1 and ECA-109 cells

Diphyllin is a natural component of traditional Chinese medicine, which effectively inhibits V-ATPase activity and affects the progression of cancer. However, few studies have been conducted on esophageal cancer, and the mechanisms remain to be elucidated. The present study revealedthat diphyllin inhibited proliferation and induced S arrest in esophageal cancer cell lines TE-1 and ECA-109. Further experiments revealed that diphyllin inhibited V-ATPase activity and decreased the mRNA expression of mammalian target of rapamycin complex 1 (mTORC1), hypoxia-inducible factor-1α (HIF-1α), and vascular endothelial growth factor (VEGF). The present study also revealed that diphyllin inhibited proliferation and reduced the formation of new blood vessels. Diphyllin inhibited blood metastasis by regulating the mTORC1/HIF-1α-/VEGF pathway, therefore it could be considered as a new V-ATPase inhibitor to treat esophageal cancer.