Establishment of heparanase stable expression cell lines

Aim:To express human heparanase in eucaryote cells and establish stable cell lines.Methods:cDNA encoding the open reading frame of human heparanase was inserted into pcDNA3.1/myc-His C.This plasmid was then transfected into CHO-K1 before stable cell lines were established by screening with G418.RT-PCR and Western-blot were applied to detect its expression;flurescent HPLC was used to evaluate its activity.Results:The CHO-K1 cells transfected with pcDNA3.1/myc-His C produced active heparanse protein,and the stable cell lines were confirm to be established.Conclusion:Heparanase stable expression cell lines have been successfully established,which could be of great benefits to the screening and function study of its inhibitors.