The Mechanism Of Dickkopf-1 Gene In Growth And Invasion Of Human Pancreatic Cancer

Objective:1. Detect the expression of DKK-1 in human pancreatic cancer cell line PANC-1;Observe the biological characteristics of PANC-1 cell after targeted inhibiting DKK-1in PANC-1 cell.2. Detect the expression changes of c-Myc,cyclinD1 and TGF-β1 in PANC-1 cell transfected with DKK-1 siRNA disturbances and analyse the correlation of DKK-1 and c-Myc, cyclinD1 and TGF-β1 to explore the correlation of DKK-1 with c-Myc,cyclinD1 and TGF-β1 in the growth and invasion of human pancreatic cancer PANC-1cell.Methods:1. 3 pairs of siDKK-1 fragment(siDKK-1-1、siDKK-1-2 and siDKK-1-3) were designed and synthed. The expression of DKK-1 in PANC-1 cell transfected with different si DKK-1 fragment was detected by RT-PCR. And the optimum jamming fragment was selected from 3 pairs of siDKK-1 fragment. The changes about biological characteristics of PANC-1 cell after blocking DKK-1 were detected from the following three aspects.The proliferation of PANC-1 cell transfected with DKK-1 siRNA disturbances was explored by MTT assay. The apoptosis of PANC-1 cell transfected with DKK-1 siRNA disturbances was detected by flow cytometry. The motility and invasion of PANC-1 cell transfected with DKK-1 siRNA disturbances was detected by the scratch test. The effects about biological characteristics in PANC-1 cell after targeted inhibiting DKK-1were explored further through comparing the differences about the three indicators in the groups.2. The expression of DKK-1, c-Myc,cyclinD1 and TGF-β1 mRNA in PANC-1 cell transfected with DKK-1 siRNA disturbances were detected by RT-PCR. The expression of DKK-1, c-Myc,cyclinD1 and TGF-β1 protein in PANC-1 cell transfected with DKK-1 siRNA disturbances were detected by western blotting.The expression change of DKK-1,c-Myc,cyclinD1 and TGF-β1 in PANC-1 cell transfected with or without DKK-1 siRNA disturbances was analysed by statistical software to explore the correlation of DKK-1 with c-Myc, cyclinD1 and TGF-β1 in the growth and invasion ofhuman pancreatic cancer PANC-1 cell.Results:1. The expression of DKK-1 mRNA was lowest after transfected with siDKK-1-1by RT-PCR.2. The rate of cell proliferation and the migration ability of PANC-1 cell transfected with DKK-1 siRNA disturbances were lower than that of blank and NC group with significant difference(Pall<0.05). The apoptosis of PANC-1 cell transfected with DKK-1si RNA disturbances was higher than that of blank and NC group with significant difference(Pall<0.05).3.The expression of DKK-1,c-Myc,cyclinD1 and TGF-b1 mRNA in PANC-1 cell transfected with DKK-1 siRNA disturbances were lower than that of blank and NC group with significant difference(Pall<0.05).The expression of DKK-1, c-Myc,cyclinD1 and TGF-b1 protein in PANC-1 cell transfected with DKK-1 siRNA disturbances were lower than that of blank and NC group with significant difference(Pall<0.05). There is positive correlation between DKK-1,c-Myc,cyclinD1 and TGF-β1 by correlation analysis.Conclusion:1.Targeted inhibiting DKK-1 in PANC-1 cell can inhibit cell proliferation and cell invasion and promote cell apoptosis.2. Blocking DKK-1 in pancreatic cancer cell PANC-1 can down-regulate the expression of c-Myc、cyclinD1 and TGF-β1.3. DKK-1 can regulate the expression of c-Myc and cyclinD1 by affecting the expression of TGF-β1 to promote the growth and invasion of pancreatic cancer cell and inhibit the apoptosis pancreatic cancer cell.