Downregulation Of P53 Gene Regulates Epithelial-mesenchymal-like Transformation Of Salivary Adenoid Cystic Carcinoma Cell Line SACC-83

Adenoid cystic carcinoma(ACC) is a common oral and maxillofacial malignancy, the incidence rate of about 10% of all salivary gland tumors accounted for 22% of all salivary gland malignancies, approximately 1% share of all head and neck malignancies. Minor salivary glands and oral is the most common disease location. ACC with perineural invasion(PNI) feature, which is a difficult clinical diagnosis and treatment of ACC. Epithelial- mesenchymal transition(EMT) refers to cells transformed by the epithelial morphology for interstitial morphology is a rapid and often reversible changes in cell phenotype has been widely introduced into the study of tumor development. p53 gene is a tumor suppressor gene, a recognized International Agency for Research on Cancer(IARC) p53 database contains more than 26,000 individual cells mutant p53 ge ne. Incidence of p53 gene mutations in different tumors, the hematopoietic malignancies p53 mutation rate of approximately 10%, ovarian cancer, colorectal cancer, head and neck cancer mutation rate of 50-70%. Found in a previous study in our group, adenoid cystic carcinoma addicted nerve invasion gene expression profiles in the presence of the p53 gene is downregulated, otherwise reported 53 gene can regulate breast cancer cell EMT process. Reveal the relationship of p53, EMT and adenoid cystic carcinoma PN I is thus important in PNI research and clinical treatment in the ACC.Objective: 1. Through RNA interference experiments proved whether p53 gene regulates EMT of ACC. 2. Experimental validation of the relationship between p53 gene and salivary adenoid cystic carcinoma PNI.Methods: 1. Using Real time-PCR, Western blot technology to screen out interference fragment of p53 gene RNA interference. 2. Downregulating p53 gene expression of salivary adenoid cystic carcinoma cell line SACC-83 cells by RN A interference technology. Then apply Western blot and flow cytometry to detect the expression of EMT typical cell surface marker, thus prove whether the p53 gene can regulate EMT of salivary adenoid cystic carcinoma cell line SACC-83 cells. 3. Using the MTT cell activity detection method to prove the impact of p53 gene expression reduction on SACC-83 cells cell viability 4. Using flow cytometry and Annexin V-FITC & PI apoptosis detection kit to detect the impact of p53 gene expression reduction on SACC-83 cells anti-apoptotic ability. 5. Using Transwell chamber cell migration assay and scratches experimental to prove the impact of p53 gene expression reduction on SACC-83 cells in vitro neurotropic migration ability. 6. Using modified Transwell chamber cell invasion assay to prove the impact of p53 gene expression reduction on SACC-83 cells in vitro perineural invasion ability.Results: 1. Successfully screened out the best fragment p53-homo-270 of p53 gene RNA interference.2. RNA interference to reduce the expressio n of P53 gene induced "EMT- like" transformation of SACC-83 cells. 3. EMT- like SACC-83 cells transformed with enhanced cell viability and anti-apoptotic ability. 4. EMT- like SACC-83 cells has a stronger in vitro PNI ability.Conclusion: 1. p53 gene may induce epithelial- mesenchymal transformation of salivary adenoid cystic carcinoma cell line SACC-83 by reduced expression; 2. p53 gene reduced expression induced EMT- like SACC-83 cell phenotype with high expression of stromal cell phenotype markers of Vimentin, N-cadherin and C-Cadherin, low expression of epithelial markers E-cadherin, EMA and CK5 which also maintain the characteristics of epithelial cell polarity; 3. p53 gene can induce EMT-like process of salivary adenoid cystic carcinoma cell line SACC-83 cells and thus involved in its in vitro perineural invasion process; 4. p53 gene can promote salivary adenoid cystic carcinoma cell line SACC-83 cells in vitro perineural invasion ability through the induction of EMT; 5. EMT- like SACC-83 cells with strong in vitro perineural invasion capacity.