Effect Of Calreticulin On Nasopharyngeal Carcinoma Cell EMT

[Objective] This project intends to investigate the expression of calreticulin in nasopharyngeal carcinoma(NPC), and further study its influence and significance of nasopharyngeal carcinoma cell EMT.[Methods] 1. Using immunohistochemical methods were used to detect the calreticulin in nasal Pharyngeal cancer CNE2 cells and nasopharyngeal carcinoma(NPC) expression,and analysis its clinical significance. 2. To construct pcDNA3.1(+) /Calreticulin eukaryotic expression vector, using cationic liposome method, transfected into NPC cell line CNE2(in pcDNA3.1(+) vector as a control), and use Western blot to analysis CNE2cells' expression of Calreticulin. 3. Use Optical microscope to observe the change on cell's form when Calerticulin is highly expressed. Through migration and invasion experiments observe high expression of calreticulin‘s influence on cell migration and invasion ability(Control with pcDNA3.1(+)); 4. The western blot detects expression of EMT related protein vimentin,MMP-9, TGF beta and E-cad expression(Control with pcDNA3.1(+)),when Calreticulin is highly expressed in nasopharyngeal carcinoma CNE 2 cells.[Results]1. Calreticulin positive rate in nasopharyngeal carcinoma was82.69%(43/52), and in nasopharynx benign tissue was 19.29%(11/57).Calreticulin expression was significantly increased(P <0.05) in nasopharyngeal carcinoma.And further analysis found that Calreticulinexpression was statistically significant correlated with the clinical stages of nasopharyngeal carcinoma and lymph node metastasis(r=0.424, P<0.05;r=0.340, P<0.05). 2.After transfection treatment, Calreticulin expression was significantly increased in nasopharyngeal carcinoma CNE2 cells.The cell morphology of CNE2 gradually evolved from a flat, cobblestone-like epithelial state into a cell long spindle fibroblast-like morphology,which shows a typical EMT phenomenon.And CNE2 cell's migration and invasion ability significantly increased than the control group(P < 0.05), the difference is significant in statistics. 3. Western blot analysis showed that,after Calreticulin increased expression in CNE2 cells, the E-cad expression significantly decreased compared with the blank control group(P < 0.05),and vimentin, MMP-9,TGF-? protein expression was significantly increased compared with the blank control group(P < 0.05), the difference is statistically significant. The results showed that high expression of calreticulin can promote the nasopharyngeal carcinoma cell line CNE2 cells undergoing EMT phenomenon,and promote migration and invasion of nasopharyngeal carcinoma CNE2 cells.[Conclusions] Calreticulin expression in nasopharyngeal carcinoma was significantly higher and positive correlation with nasopharyngeal carcinoma stage and lymph node metastasis.Calreticulin can enhance migration and invasion of nasopharyngeal carcinoma CNE2 cells by promoting EMT phenomenon.