fascinser39促进食管癌细胞侵袭移动的作用机制及fascin新相互作用蛋白的鉴定word格式论文.docx

优秀毕业论文 精品参考文献资料 Abstract Fascin, an actin bundling protein, has been more and more concerned because of its roles in invassivness and metastasis in carcinoma cells. It has been researched that Fascin did not expressed in normal human epithelia tissue or low expressed in basal layer cells. Fascin overexpressed in many malignant carcinoma cells and it has been associated with poor prognosis and more metastasis. It has been reported that Ser39 of Fascin is a high conservative phosphorylation region, located in the actin bind site 1. And the phosphorylation of this site could inhibit the actin bundling activity of Fascin. The identification of protein-protein interaction of Fascin is a very important scientific issue and it will facilitate to elucidate the mocecular mechanism of initiation and progression of malignant carcinoma. But, so far, the identification of interaction with Fascin has rarely been reported. In view of this, we plan to study the role of phospharylation of Fascin/Ser39 in ESCC, and intend to identify a novel protein-protein interaction with Fascin, based on the result that Fascin promote the abilities of invasiveness and motility in ESCC cells which our group ever researched. Description as follows: First, we inserted the whole coding sequence of fascin into pEGFP vector, constructing pEGFP-fascin expression vector, and then we constructed two expression vectors containing fascin mutants at fascin/Ser39 site, pEGFP-fascin/S39A and pEGFP-fascin/S39D. Second, pEGFP, pEGFP-fascin, pEGFP-fascin/S39A and pEGFP-fascin/S39D have been transient transfected into KYSE150 cells to examine the difference of cancer cell’s abilities of motility and invasiveness, and observe the distribution of F-actin. The result shows, 1) the abilities of motility and invasiveness of the KYSE150 cells after transfected WT were increased and the spinous protrusions bolstered by F-actin also increased, compared to cells transfected vector, and F-actin colocalized with Fascin; 2)