Regulation Of Pokemon On Cyclina In Hepg2 Cell

Objects: To Investigate the role of proto-oncogenes Pokemon in regulating the cell c-ycle proteins CyclinA, which can provide the foundation for the further study.Methods: In this study, we first construct two CyclinA promoter luciferase repo- rter plasmid with different length pLuc-300 and pLuc-600, and then co-transfect into HepG2 cell with pcDNA3.1-Pokemon over-expressed plasmid and pcDNA3.1-empty plasmid. After 48 hours, detect the CyclinA promoter's transcriptional activity by usi- ng D-ual-Luciferase Reporter Assay;Finally, transfect the HepG2 cell with Pokemon siRNA and Scramber siRNA, harvest the cell at 0, 24, 48, 72 hours, detect the protein expression level of Pokemon and CyclinA by Western-Blot.Results: Dual luciferase reporter assay confirmed that both of pLuc-300 and pLu c-600 had transcriptional activity, but the activity of pLuc-300 was much more stron- ger than pLuc-600. Pokemon could repressed the transcriptional activity of pLuc-300 and pLuc -600. Down- regulating Pokemon protein expression level by siRNA could incr-ease the protein level of CyclinA with a time dependent manner.Conclusions: Pokemon can inhibite the transcriptional activity and protein expre- ssion level of the CyclinA in HepG2 cell.