Effect Of PHD2 Gene Silencing On Proliferation,Metastasis And Vasculogenic Mimicry Of Prostate Cancer Cell Line PC-3

Objective:To examine the role of PHD2 gene in Prostate cancer Vasculogenic mimicry process and then study its effect on PC-3 cell proliferation,migration and invasion ability.Methods:1.First,human prostate cancer line PC-3 were cultured in vitro,at the same time,4 PHD2-shRNA sequences ware transfected to silence PHD2 gene in prostate cancer PC3 cell,the silencing efficiency was deteceted by Western blot and RT-PCR,and the effective interference group was selected for the experimental group.2.The experiment was divided into PC3 Group,PC3+shRNA-NC Group and PC3+PHD2shRNA group.The protein expression of PHD2 and HIF-1α and EphA2 were delected by Western Blot of each group after transfection.3.Each group of PC3 cells was cultured in vitro with 3D condition to observe the number of vasculogenic mimicry in PC3 cells of each group.4.The changes of proliferation,migration and invasion ability of PC3 cells in each group were detected by MTT multiplication experiment,cell scratch experiment and Transwell invasion experiment.Result1.The results of the Western blot and RT-PCR experiments showed that the mRNA expression level and protein expression level of PHD2 in PC3 cells decreased significantly after four groups of shRNA plasmid were transfected to PC3 cells,and the difference was statistically significant(P<0.01).Obtained the experimental group PHD2-shRNA sequence,the silence efficiency reached more than 80%.2.The vitro three-dimensional culture experimental results show that:in vitro three-dimensional culture 48 h,compared with the blank group and negative control group,PC3 cells of PHD2 silent group can see more VM sample structure,the difference have statistically significant(P<0.05).The results suggest that silent PHD2 can promote the formation of VM.3.MTT multiplication experimental results show that the cell activity detection at different time(24h,48 h,72h),compared with the blank group and the negative control group,the activity of PC3 cells in PHD2 silent group increased time-dependent,the difference was statistically significant(P<0.05).4.Scratch experimental results showed that: to observe the migration distance of each group of cells at different time points(24h,48 h,72h),the PHD2 silence group and the negative group and the blank group found that the migration distance of PHD2 silent group cells was significantly larger than that of blank group and negative group in different time and the difference was statistically significant(p< 0.05).5.The results of the Transwell experiment showed that compared with the blank group and the negative group,the number of cells entering the lower Transwell chamber in the PHD2 silent group increased significantly,and the difference was statistically significant(P<0.05).ConclusionThe expression of PHD2 in PC3 cells of prostate cancer through RNA interference technique can promote the proliferation,migration,invasion and the formation of vasculogenic mimicry of prostate cancer PC3 cells by increasing the expression of HIF-1α and EphA2.