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Objective To study the effect of human insulin gene modified fibroblasts on blood glucose in diabetic rats.Methods An expression vector containing human insulin gene was constructed by recombinant DNA technique and introduced into fibroblast Ltk cells by lipofectin-mediated DNA transfection. Following G418 screening, the survived cells were selected and enriched. Finally a cell line, PRI-12 was generated for the highest insulin production. Then, these cells were injected into streptozocin (STZ)-induced diabetic rats.Results insulin DNA transfected Ltk- cells were able to express insulin at a high level in a long-term culture. Furthermore, these Ltk- transfectants could decrease blood glucose significantly (P<0.01) and obviously increase the body weight (P<0.01) when they were injected into STZ-induced diabetic rats.Conclusions These results suggest that the cell lines transfected insulin gene could secrete insulin and execute effect on diabetic rats. The study provides support for the view that somatic cell gene therapy offers a potential approach to delivery insulin into diabetes mellitus.