【Abstrat】 Objective To stably express human erythropoietin (EPO) gene in human proximal tubular epithelial cells (HK-2). Methods Eukaryotic expression vector for human EPO gene was constructed by the standard molecular cloning method. The recombinant pcDNA3.1(-)-hEPO vector was transfected to 293 cells and HK-2 cells by the calcium phosphate coprecipitation method. EPO in cultured supernatant was detected by ELISA. Proliferation of HK-2 cells was assayed by MTT method. Results After successful construction of the recombinant plasmid and transfection of the plasmid to HK-2 cells, EPO was temporarily expressed in transfected HK-2 cells, and was constantly expressed in transfected HK-2 after selection by G418. The expression level of EPO in transfected HK-2 cells was 35.0±5.3 % of that in transfected 293 cells. HK-2 cells stably expressing transfected human EPO did not show any changes of cell growth. Conclusion Human EPO cDNA can be stably expressed in human proximal tubular epithelial cells (HK-2), suggesting a promising approach for EPO gene therapy.