A Study On The Effect And Mechanism Of The New Tumor Suppressor CYLD

Objective:To Construct the pcDNA3.1(-) /CYLD expression plasmid and investigate its stable expression in Breast cancer Bcap37 cells, which we can study the function of CYLD that palys in breast cancer Bcap 37 cells.Methods:CYLD gene was bought from the foreign,s gene bank,and The eukaryotic expression vector pcDNA3.1 (-)/CYLD was constructed by Shanghai Chemical Technology Co.Ltd. The recombinant pcDNA3.1( -)/CYLD was sequenced and transfected by lipofectamine into Breast cancer Bcap37. Through G418 filtration ,We obtained the positive Breast cancer Bcap37 cells. MTT , clone formation and western Blot were done to detect the proliferation and protein expression in vitro.Results:The eukaryotic expression vector pcDNA3.1(-)/CYLD was constructed and transfected successf lly in Breast cancerBcap37 cells. The result of MTT showed A490nm lowered,so the growing Breast cancer of Bcap37 was inhibited. The ratio of growth inhibition is about 72.5% after 7 days accoding to growth curve test.The experiment of clone formation showed the rate of clone formation reduced,and have statistics significance compared with untransfected cells (P <0.05).Western Blot demonsrated the inceased protein expression.Conclusion:The eukaryotic expression vector DNA3.1(-) /CYLD ,which can be expressed stablely in Breast cancer Bcap37 ,has been successfully transfected .CYLD Negatively regulate the biological behavior of the Breast cancer Bcap37. This study laid a foundation for the further research o f the CYLD and there will be the potential to provide new gene therapy targets for breast cancer.