Up-regulation Of MicroRNA-101 Inhibits Prostate Cancer Cell Metastasis By Modulating Expression Of CRMP4

Objectives:Investigate the correlation among mi R-101, EZH2 and CRMP4 in prostate cancer cell; make sure whether mi R-101 in prostate cancer modify histone of CRMP4 promoter by regulating EZH2, further clarify epigenetic regulatory mechanisms of CRMP4. Methods:Prostate cancer PC3 cell were transfected with pre- mi RN A-101 mimic 、 pre-mico RN A negative control,si RNA EZH2,si RNA negative control 、si RNA positive control(si RNA MAPK-1) by using liposome Lipofectamine 2000.To observe fluorescence brightness of tall group by fluorescence microscopy.The efficiency of transfction was tested by real-time RT-PCR; m RNA expression of CRMP4 and EZH2 was detected by real-time RT-PCR。PC3 cells were treated with 3-Deazaneplanocin A(DZN EP),which will inhibit the histone methyltransferase activity of EZH2 as control.Protein expression of CRMP4 、EZH2、 H3K27me3、H3A and u H2AK119 were detected by western-blot。 Results:6 hours after transfection,there is bout 85% of the cells have fluorescence in negative control.Real- time RT-PCR results showed that micro RNA-101 was significantly up-regulated after transfection for 48 hours,C RMP4 m RNA increased significantly and EZH2 m RNA decreased significantly,.Westrn blot results showed that CRMP4、H3AC protein increased significantly, EZH2 、H3K27me3、u H2AK119 protein decreased significantly。 Conclusion:Upregulate the micro RNA-101 can inhibit EZH2 m RN A and protein expression,thus down regulate H3K27me3、u H2AK119 protein expression,upregulate H3 AC protein finally upregulate CRMP4 m RNA and protein expression。...