The Correlation And Mechanism Between NDRG1 Gene And The Biological Behavior Of Pancreatic Cancer

Objectives:To investigate the expression of NDRG1 in pancreatic cancer tissues and adjacent non-cancer tissues as well as its mechanism in pancreatic cancer development.Methods:1)By RT-PCR?Western Blot,we analyze mRNA and protein of NDRG1 in pancreatic cancer and adjacent non-cancer tissues.Then according to the expressions of NDRG1 in pancreatic cancer cell lines,Mia PaCa2 and PANC-1 were used to in vivtro experiments.2)NDRG1 shRNA and vector were designed to upregulate or inhibit NDRG1 in pancreatic cancer cell lines.Stable cell lines were identified by RT-PCR?Western Blot.3)The motility of pancreatic cancer cell was tested by invasion,migration and scarification experiments.The proliferation,apoptosis and cell cycle were analyzed by CCK-8,PI staining and Annexin V-FITC/PI methods respectively.The expression of STAT3?MMP-2?MMP-9?PTEN?p-AKT?Caspase3 were identified by western blot after stable cell lines.Results:1)In 14 pairs pancreatic cancer tissues and adjacent non-cancer tissues,the mRNA of NDRG1 is different:the expression of NDRG1 in 12 cancer tissues is less than adjacent non-cancer tissues.And the protein of NDRG1 in cancer samples are all less than that of adjacent samples.In 10 pancreatic cancer cell lines,NDRG1 is lowly expressed and thus selected to subsequent experiments.2)We design and construct vector of NDRG1 and acquired the stably upregulated NDRG1 as well as downregulated NDRG1 cells.3)In invasion,migration and scarification experiments,we find that overexpressing NDRG1 inhibit pancreatic cancer cells motility.In proliferation experiments,the number of upregulated NDRG1 Mia PaCa2 and PANC-1 is significantly less than control cells.Overexpressing NDRG1 also increase the number of panc1 cells in S stage and apoptosis in Mia Paca2 cells.Western blot experiments reveal that in upregulated NDRG1 pancreatic cancer cells,STAT3?MMP-2?MMP-9?p-AKT are lower than control but PTEN?N Caspase3 are higher than control and vice versa.Conclusion:1)The obersevations suggest that NDRG1 is tumor suppressor gene in pancreatic cancer,which may be associated with low expression of poor differentiation,high invasion of pancreatic cancer.2)The successful establishment of stable overexpression and silencing NDRG1 pancreatic cancer cell lines,is ready for experiments in vitro and mechanisms of NDRG1 pancreatic cancer.3)The overexpression of NDRG1 can induce apoptosis in pancreatic cancer cells,inhibition of cell proliferation and invasion and metastasis;silencing of NDRG1 can promote invasion and metastasis even more proliferation in pancreatic cancer cells.4)NDRG1 may regulate STAT3 signaling pathway,which reduced the expression of MMP-2 and MMP-9,result as inhibiton of pancreatic cancer invasion and metastasis.On the other hand,NDRG1 can regulate the expression of PTEN,p-AKT,Caspase 3 protein,lead to inhibition of cell proliferation and induce apoptosis.Its low expression might be associated low differentiation and high invasion ability.It plays a key role in the development of pancreatic cancer.