The Effect Of Silencing UBE3A Gene On Biological Behavior Of Salivary Adenoid Cystic Carcinoma Cell

Salivary adenoid cystic carcinoma（SACC）is a common malignant tumor that occurs in the salivary glands.Various methods have been tried in order to control this disease,but the results are not satisfied.Therefore,we urgently need to find a new effective treatment method.Ubiquitin-protein ligase E3A（UBE3A）is one of the important members of the ubiquitin-proteasome system（UPS）.UBE3 A has been confirmed to be involved in the occurrence and development of a variety of tumors.The previous studies of our research team have confirmed that the expression of UBE3 A in SACC tissue is significantly higher than that of normal salivary gland tissue.Therefore,whether UBE3 A can become a potential target of SACC gene therapy is worthy of further exploration.Objective:1.In order to explore the influence of UBE3 A on SACC cells invasion and migration,we tested their invasion and migration ability after UBE3 A silence.2.In order to explore whether UBE3 A is involved in the proliferation and apoptosis of SACC cells,we tested their proliferation and anti-apoptotic ability after UBE3 A silence.3.In order to explore the possible mechanism of UBE3 A affecting the biological behavior of SACC,we tested the changes in the expression levels of VEGF,p53,and PTEN proteins in SACC cells after UBE3 A silence.We hope that this result will lay the foundation for studying the function and mechanism of ubiquitination in SACC.Methods:A recombinant lentivirus that silences UBE3 A gene expression was constructed and stably transfected into SACC-LM cells.The highest silencing group is uesd in the following assay.We used both RT-q PCR and Western blot to detect the expression of UBE3 A in cells at the gene and the protein level.In order to verify the difference between normal and silence cells,then,we used transwell assays,wound healing,CCK-8 and flow cytometry methods to detect cell invasion,migration,proliferation and apoptosis.Western Blot was also adopted to detect the expression of VEGF,p53 and PTEN.According to the comparative analysis of the experimental results,we have discovered the effect of silencing UBE3 A gene on biological behavior of salivary adenoid cystic carcinoma cell.This study lays the foundation for UBE3 A to become an effective therapeutic target for SACC.Results:The experimental results of transwell assays,wound healing and CCK-8 respectively showed that the invasion,migration and proliferation of SACC cells were inhibited after UBE3 A silence.The results of flow cytometry showed that the apoptosis of SACC cells increased after UBE3 A silence.The experimental results of Western Blot showed that after the expression of UBE3 A was down-regulated,the expression levels of p53 and PTEN protein were up-regulated,while the expression of VEGF was down-regulated.Conclusion:The overexpression of UBE3 A in human salivary gland tissue may be involved in the occurrence of SACC,and promotes tumor cell migration,invasion,proliferation and anti-apoptotic ability during tumor development.Our research shows that preventing the occurrence and development of SACC by inhibiting the expression of UBE3 A may be a potential treatment method.