The Effects And Clinical Pathological Significance Of TWSIT1 Gene On Esophageal Cancer Associated Fibroblasts

Objectives:To detect the influence of conditioned medium of esophageal cancer associated fibroblasts which express Twist1 gene on migration and invasion abilities of esophageal squamous cell carcinoma (ESCC). To further discuss the relationship of Twist1 expression and ESCC clinicopathological feature and the patient survival rate. Providing the effective target and making a good predictor of prognosis for gene therapy of esophageal cancer.Methods:Fibroblasts were isolated and cultured from normal esophageal mucosa, made them differentiate into cancer associated fibroblasts with different concentration of IL-6,The expression of Twist1 was detected by Western-blot and Fluorescence quantitative RT-PCR. To detect the influence of conditioned medium of esophageal cancer associated fibroblasts which express Twsitl gene on migration and invasion abilities of esophageal carcinoma cells by wound healing assay、Transwell migration and invasion assay;Twistl expression in ESCC interstitial tissue was detected by immunohistochemisty, to analyze the relationship of Twistl expression with cancer associated fibroblasts markers as well as the clinical pathological features and the survival rate of patients with ESCC.Results:After primary cultured normal esophageal fibroblast cells were treated with IL-6, the expression level of the Twist1、SMA、FAPa increased. Added the conditioned medium to the TE8 esophageal carcinoma cell lines, compared with the control group (Without the condition medium group or normal fibroblast conditioned medium group), the faster scratches healing.the number of cells migration and invasion from the Transwell chamber increased significantly. Twistl expression in ESCC interstitial tissue was closely correlated to the expression of cancer associated fibroblast markers, such as FAP, PDGFRa, PDGFRβ, SMA. And as well as T stage, clinical stage, Lymph node metastasis of the ESCC. In esophageal fibroblasts, Twistl positive group showed significantly lower overall survival (OS) and disease-free survival (DFS) than negative group.Conclusion:Conditioned medium of esophageal fibroblasts with expresses Twsitl gene can promote migration and invasion of esophageal carcinoma cells. Twistl expression in stromal fibroblasts of ESCC will be helpful determine and assess the prognosis of patients. Twistl may be used as the esophageal cancer associated fibroblasts marker, and provide an effective gene target for therapy of esophageal cancer.