Expression Of SIRT3 In Pancreatic Cancer And Its Clinical Significance

ObjectiveTo observe expression of SIRT3 in the pancreatic cancer and analyze its significance.Build SIRT3 cut the stability of the protein expression of pancreatic cancer cell line,observe the apoptosis and appreciation ability and the relevant affect protein expression in pancreatic cancer cells.MethodsImmunohistochemistry and Western-blot was performed to investigate the expression of SIRT3 in pancreatic cancer tissue and its expression in normal pancreatic tissue,draw the curve of patients with pancreatic cancer growth,and analyze the expression level of SIRT3’s relationship with the clinical pathological features.Using Real Time PCR to detect SEPAC-SIRT3 mRNA expression in five kinds of pancreatic cancer cell line,and choose SIRT3 mRNA expression quantity highest as building SIRT3 protein expression by pancreatic cancer cell lines.Using SiRNA interference SIRT3 protein expression,constructing SIRT3 protein expression decreased the stability of the pancreatic cancer cell lines.Using flow cytometry instrument testing before and after the disturbance the apoptosis of pancreatic cancer cell line capacity.MTT method is used to detect pancreatic cancer cell line proliferation ability before and after the interference.The Western-blot was used for the test related affect protein expression in pancreatic cancer cells before and after the interference.Results1,Test results by Immunohistochemistry method and Western blot showed that positive expression rate of normal pancreatic tissue was significantly higher than that of pancreatic tissue(P<0.01).2,SIRT3 expression level of high or low in the pancreatic cells,had no relationship with gender,age,tumor diameter,pathology types,lymph node metastasis and TNM stages.In patients with and without vascular invasion,the difference was significant(P < 0.05).3,Patients survival time in SIRT3 positive expression were longer than patients with negative expression.4,PANC 1 was chosen as the object of cell transfection.Three stable cell line were got,and # 2,# 3 cut were more apparent,so use these two as the follow-up study.3,2 mu M staurosporine treatment of pancreatic cancer using si-Ctrl,# 2,# 3,were determined by MTT test,the results showed that the staurosporine apoptosis of pancreatic cancer si-Ctrl was significantly higher than # 2,# 3(P < 0.05).4,Pancreatic cancer cells si-Ctrl and # 3,72 h,at 0 h,48 h and 72 h were determined by MTT test,0 h cell vitality,no difference in both groups(p > 0.05),48 h and 72 h,# 3pancreatic cancer cell proliferation activity was significantly higher than that of si-Ctrl(p< 0.05).5,Using Western blot to detect the related invade protein in si-Ctrl,#2,#3,it is found that except MMP7 other level of attacks protein is higher in #2,#3 than in si-Ctrl.ConclusionSIRT3 present was high expression in normal pancreatic tissue;SIRT3 low expression in pancreatic cancer tissue.SIRT3 protein can promote apoptosis of pancreatic cancer,and the ability to inhibit pancreatic cancer cell proliferation activity.SIRT3 can lead to the decrease of the related affect protein expression in pancreatic cancer cells.