Effects Of SiRNA Silencing DAD1 Gene Expression On A498 Cell Proliferation And Invasion Of Renal Carcinoma

Background:Renal cell carcinoma (also known as renal cell carcinooma RCC) is the most common malignant tumor of adult kidney, malignant tumor accounted for adult 2%-3%, respectively ranked seventh in males and ninth in females in malignant tumor. In the scope of the global, about 270000 new cases each year, including 120000 deaths, in China, the incidence of renal cancer after bladder cancer in urologic tumors, and in the past 20 years by 6% every year.The pathology of renal cell carcinoma including:clear cell carcinoma, papillary renal cell carcinoma, chromophobe cell carcinoma, collecting duct carcinoma and undifferentiated carcinoma.All of which,the clear cell carcinoma (clear cell renal cell carcinoma ccRCC) is the most common histological type, originated in the epithelial cells of the proximal tubules, and prone to metastasis, accounting for metastatic renal cell carcinoma 80%-85%.RCC are not sensitive to radiotherapy and chemotherapy, surgical operation is the best treatment method, but the recurrence rate is about 20%-40%.Because renal cancer chemotherapy and radiotherapy resistance, hence it is no effective postoperative treatment.As one of the common diseases in urological department,renal cell carcinoma, especially the early renal cell carcinoma of no special clinical manifestation, to appear in the urine, low back pain or abdominal mass when is terminal.Renal cell carcinoma are not sensitive to radiotherapy, chemotherapy, I clinical generally treated by radical nephrectomy, but once the lymph node metastasis, even for radical lymph node dissection, the survival of patients with little more than 5 years, if the occurrence of liver and lung metastasis or adjacent organ invasion is a worse prognosis, so finding out a new treatment option becomes more and more important.Currently, targeting silence target gene to treat cancer is paid attention to by more and more researchers, according to the researches of anti apoptosis gene silencing in tumor cells can promote greatly the apoptosis of tumor cells, play a role in the treatment of carcinoma. Objective:To observe the effect of RNAi-mediated silencing of the DAD1 gene on the proliferation and invasion of A498 cell.Methods:Three siRNA targets toward DAD1 gene were designed and transfected to RCC cell line A498,and universal scrambled negative control siRNA group (NC group). The gene expression of DAD1 were determined using real-time quantitative PCR and western blotting. The cell viability were determined by MTT assay. The Annexin V-FITC/PI double-labeled flow cytometry was employed to measure the apoptosis at 48h after transfection. Transwell migration assay were used to evaluate migration of trans-fected cells.Results:Real time PCR and western blotting test results showed that three DAD1-siRNAs could effectively inhibit DADl mRNA and protein expression(P<0.05), which DAD1-siRNA1 silence has the most significant effect(P<0.05); MTT assay result displayed the cell growth and proliferation were inhibited obviously when interferenced target effectivly was transfected 24,48 and 72 hours(p<0.05); Transwell test results showed that transfection DAD1-siRNA and untransfected negative control siRNA invasion irradiated group were significantly different(p<0.05);Conclusion:DAD1 siRNA could efficiently inhibit DAD1 gene expression and the cell proliferation,induce the tumor cell apoptosis and repress the invasion in RCC cells.