The Inhibitiory Effects Of Interferon-gamma On The Migration, Invasion, And Vasculogenic Mimicry Formation Of Human Prostate Cancer Cell Line PC-3

Objective:To investigate the effects and mechanism of interferon-gamma(IFN-?)on the migration,invasion,and vasculogenic mimicry(VM)formation of human prostate cancer cell line PC-3.Methods: Human prostate cancer cell line PC-3 was used as the research object.The cells were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum(FBS),the well-grown PC3 cells were divided into the control group(only added with RPMI1640 medium)and different concentration of IFN-? treatment group(10,100,1000 ug/L).The invasion and migration of PC3 cells were determined by the wound-healing assay and cell invasion assay,respectively,and the formation of VM in PC3 cells was observed by three-dimensional cell culture.The m RNA expression level of erythropoietin –producing hepatocellular carcinoma-A2(EphA2)was detected by RT-RCR,and the protein level of EphA2 was detected by Western Blot.Results:1.Wound-healing assay showed that different concentrations of IFN-?(10?100?1000?g/L)treatment shortened the migration distance in human prostate carcinoma PC3 cells at different time points(24?48?72 h),and the migration distance of human prostate carcinoma PC3 cells was shortened by IFN-? with a concentration-dependent manner,indicating that IFN-? inhibited the human prostate carcinoma PC3 cells migration ability(p < 0.05).2.Transwell invasion experiment showed that 24 h after IFN-?(10?100?1000?g/L)treatment,the number of invasive cells were significantly decreased compared with the control group(p < 0.01),indicating that IFN-? inhibited the human prostate carcinoma PC3 cells invasion ability(p < 0.05).3.Three dimensional cell cultures results showed that the human prostate carcinoma PC3 cells with more VM channels in the control group,while the IFN-? treatment significantly reduced the formation of VM channels at the concentrations of 10?g/L,and few VM channels could be found in the human prostate carcinoma PC3 cells after IFN-? treatment at the concentrations of 10?g/L and 1000?g/L,which indicates that IFN-? suppressed the formation of VM channels in the human prostate carcinoma PC3 cells.4.The RT-PCR results showed that IFN-? treatment decreased the m RNA expression level of EphA2 with a concentration-dependent manner in human prostate carcinoma PC3 cells(p < 0.05 and p < 0.01).5.Western blot results showed that IFN-? treatment the protein level of EphA2 with a concentration-dependent manner in human prostate carcinoma PC3 cells(p < 0.05 and p < 0.01).Conclusion: IFN-? inhibits the migration,invasion,and VM formation of human prostate carcinoma PC-3 cells,involving the suppression on m RNA expression and protein levels of EphA2.