NDRG1 up-regulation influences proliferation and apoptosis of human pancreatic cancer cells
[Objective] To observe the effect of N-myc downstream regulated gene-1 (NDRG1) up-regulation on proliferation,cell cycle and apoptosis of pancreatic cancer cells and discuss the mechanism.[Methods] Western blot was used to detect the NDRG1 expression in PANC-1,BXPC-3,CAPAN-2 and SW1990 cells.Over-expression plasmid pEGFP-NDRG1-N3 was constructed and transfected into CAPAN-2 cells to up-regulate the expression of NDRG1.Interfering effect was detected by immunofluroscence,Western blot and RT-PCR.Proliferation,cell cycle and apoptosis were detected by MTT,PI method and flow cytometry respectively after transfection.[Results] Western blot showed that NDRG1 was expressed in PANC-1,BXPC-3,CAPAN-2 and SW1990 cells,the expression was higher in the poorly-differentiated cells (PANC-1) than in the moderately-differentiated (BXPC-3) and well-differentiated (CAPAN-2,SW1990)cells (P ＜ 0.05).Immunofluroscence showed that the positive fluorescence pEGFP-NDRG1-N3 cells were over 70％ and NDRG1 was up-regulated at protein and mRNA levels.Parp,Cleaved Parp,p-P53 and Cleaved-Capase 3 were not changed at mRNA level after NDRG1 up-regulation (t=1.456,1.164,2.914 and 1.075;P＞0.05).At protein level,Cleaved Parp was up-regulated,while Parp,p-P53 and Cleaved-Capase 3 were down-regulated (t=12.273,6.104,3.691 and 14.227;P＜0.05).MTT showed that compared with the pEGFP-N3 group,the CAPAN-2 cell proliferation in the pEGFP-NDRG1-N3 group was increased at 96 and 120 h (t =8.176 and 2.246,P＜ 0.05).PI method showed that compared with the pEGFP-N3 group,CAPAN-2 cells in G1 phase were significantly increased while those in G2 and S phases were significantly decreased in the pEGFP-NDRG1-N3 group (t=3.651,4.133 and 3.092;P＜0.05).Flow cytometry showed that the apoptosis rate in the pEGFP-NDRG1-N3 group was lower than that in the pEGFP-N3 group (t=9.161,P＜ 0.01).[Conclusions] NDRG1 up-regulation in pancreatic cancer cells can increase the proliferation,inhibit apoptosis and promote cells staying in G1 phrase.NDRG1 may be a targeting candidate for pancreatic cancer therapy.
国家科技期刊平台
NSTL
万方数据
维普
