Investigation Of WASH Promotes Esophageal Squamous Cancer Progression

Background and purpose As one of common malignancies,esophageal cancer threstens seriously the health of people.The morbidity and mortality of esophageal cancer rank first in the world,which causes killing over 150,000 people annually.In China,the 5-year survival rate for esophageal cancer patients is less than 20%.The incidence of esophageal cancer is high among male and mainly afflicts people over 40.In recent years,we found that the incidence of esophageal cancer increased significantly,so elucidating the pathogenesis and developing new drug of esophageal cancer are imminent.For the esophageal squamous cell carcinoma in China accounted for 90%of esophageal cancer,this study mainly focuses on the research of esophageal squamous cell carcinoma.More and more reports show that the cytoskeleton has far more biological significance than simple a concept of cell morphology.It has been found that the cytoskeleton is involved in almost all the activities of vital cell life,such as intracellular Spatial loci,cell movement and signal transduction.Recently,it has been founded that the variation of structure and function of the skeletal structure plays an indispensable role on the onset and the development of cancer.And many studies have shown that the malignant cellphenotype of tumor cells usually associated with abnormal expression of actin-associated protein.In this study,we are concerned about the cytoskeletal regulatory protein WASH,also known as WASHC1,which is an important cytoskeletal regulatory protein of the WASP(Wiskott-Aldrich syndrome Protein)family.And researches about the WASH cytoskeletal regulatory gene involved in the development of the tumor are still rarely reported.As an important cytoskeletal protein,the issues on whether WASH can affect the fate and the progress and metastasis of tumor cells are worthy of further study and research.The main tasks of this topic are identifying the important role of WASH on the occurrence and developmen of esophageal squamous cell carcinoma,finding the mechanism on the WASH how to affect the esophageal cancer progression,and exploring the significance further combined with the clinical.Result1.The results of fluorescence quantitative PCR showed that the expression of WASH was higher than that in adjacent tissues,and the difference was statistically significant.WASH expression and pathological staging have a significant correlation.2.The results of immunohistochemistry showed that the expression of WASH was higher than that in adjacent tissues,and the difference was statistically significant.WASH expression and pathological staging and differentiation were significantly correlated.And WASH was negatively correlated with patient prognosis.3.The results of sphere formation experiments showed that after silencing WASH,the number of sphere formation of esophageal squamous carcinoma cells and the expression level of the stemness gene were significantly decreased.After transient overexpression of WASH,the number of sphere formation and the expression level of stemness genes increased were significantly increased.4.The results of multi-cytokine test showed that the expression of cytokine CXCL8 was significantly decreased after silencing WASH.The differences were statistically significant.5.Animal experiments showed that tumor volume and quality in the group of subcutaneous injection of silencing WASH cell lines was significantly reduced compared with the control group.In the tumor tissue of mice,the expression of CXCL8 was significantly decreased compared with the control group.6.The 3'-UTR of mi R-637 targeting WASH was detected by targetscan,mi Rbase and other predicted sites and using dual fluorescence reporter assay.And after transient transfection of mi R-637,the expression of WASH was significantly reduced.And by constructing a stable overexpression system of mi R-637,the expression of WASH was found to be significantly reduced.In 45 cases of esophageal squamous cell carcinoma,the expression of mi R-637 in the adjacent tissue is higher than that in the cancer tissue,the difference was statistically significant.In clinical patients,mi R-637 expression was significantly negatively correlated with WASH expression.Conclusion1.WASH is highly expressed in esophageal squamous cell carcinoma.WASH enhances the stemnesss of esophageal squamous cell carcinoma and promots the development of esophageal squamous cell carcinoma.2.In esophageal squamous cell carcinoma,low expression of mi R-637 leads to WASH up-regulation,then the up-regulation of WASH leads to the promoting of CXCL8,and the above-mentioned factors results in enhancing the stemness of esophageal squamous cell carcinoma and promoting esophageal squamous cell carcinoma progress.