Establishment of human heparanase stable expression CHO cell lines

Objective To transfect human heparanase( HPA) gene into CHO cells and establish the stably expressed cell lines. Methods The recombinant plasmid p EGFP-N1-HPA was transfected into CHO cells by lipofectmine2000 and the stably expressed cell lines was selected by G418. Fluorescence microscope was used to observe the expression of green fluorescence. RT-PCR and immunocytochemistry were applied to detect the HPA expression at transcription level and protein level,respectively. The proliferation of transfected CHO cells was measured by the MTT method. Results The transfected CHO cells expressing green fluorescence were observed by fluorescence microscope. HPA gene was also determined in the stably transfected cells.Conclusion The CHO cell line with stable expression of human heparanase has been successfully established,which may provide a great benefit for study the function of this enzyme.