The Inhibition Of HES1 And HEY1 Effects On Salivary Adenoid Cystic Carcinoma Cell Function

Salivary adenoid cystic carcinoma(SACC), one of the common malignant tumors in oral and maxillofacial region, usually leads to postoperative recurrence and poor prognosis. Its clinical features are feeling and motor nerve dysfunction, and also easy to transfer to the lungs, because of perineural invasion and distant metastasis. Our previous study has demonstrated the close relationship between Notch pathway and SACC and its receptor gene Notch1 contributed to proliferation, invasion and metastasis of SACC. Numerous researches had veritified that HES1、HEY1 were the downstream target genes of notch signaling pathway, which might function as oncogene in many tumors development. The purpose of this study was to screen out HES1、HEY1 and explore their effects on the proliferation, invasion and metastasis of SACC cells. This research was divided into two parts as followed.1. Screen out the downstream target genes of Notch1 with real-time PCR and demonstrate its expression on the level of organizationAim: To screen out the common downstream target genes of Notch1Methods: Select several common downstream genes of Notch1 signal pathway based on literature and design the primer sequences. Design and synthesize si RNA- Notch1 and transfect SACC-83 cells to study the changes of its downstream genes with real-time PCR. Construct the sh RNA recombinant adenovirus vector and infect SACC-83 cells to study the changes of target genes with real-time PCR. Utilize immunochemistry to evaluate the results of reaction with target antibody in fresh SACC excision samples.Results: The results of real-time PCR revealed that the expression of HES1、HEY1 downregulated with Notch1 being inhibited as well as HES1、HEY1 upregulated with Notch1 being overexpressed. The expression of HES1、HEY1 in carcinoma was much higher than in para-carcinoma.Conclusion: The expression changes of HES1、HEY1 were positive correlation with Notch1.2. Using si RNA to verify the effects of HES1、HEY1 on cell function of salivary adenoid cystic carcinoma.Aim: To verify the effect of HES1、HEY1 on the proliferation, metastasis and invasion of SACC cells.Methods: At first, si RNA-HES1 and si RNA-HEY1 were synthesized and transfected into SACC-83 cells, then their expression was examined by real-time PCR. The proliferation of SACC-83 cells was tested by CCK8 and plate cloning assays. Transwell and wound healing assays were used to evaluate cell metastasis and invasion. Flow cytometry was applied to investigate the changes of cell apoptosis and cell cycle.Results: The results of real-time PCR revealed that the target genes expressed less. The growth of SACC-83 cells transfected with HES1-si RNAs and HEY1-si RNAs was significantly suppressed in CCK8 and plate cloning assays. After the expression of HES1、HEY1 decreased, the metastasis and invasion ability of SACC-83 cells were verified suppressed in transwell and wound healing assays. The data and diagram of flow cytometry showed that the early and late apoptosis rates rose but the cell cycle seemed insignificance.Conclusion: HES1、HEY1 can inhibit the proliferation, invasion and metastasis of SACC-83 cells.