Objective  To construct an “artificial beta cell” that possesses mature insulin secretion from a human hepatoma cell line HepG2.   Methods  The recombinant retroviral vector pL-mhPINS-SN was transfected into PA317 packaging cells and selected with G418 selective medium. Virus titer was detected with NIH3T3 cells

and PA317/mhPINS cells were assessed with PCR. HepG2 cells were infected by mhPINS virus and screened with G418. HepG2/mhPINS cells were tested with RT-PCR

Western blot and

radioimmunoassay

respectively. The insulin secretion stimulated by glucose in HepG2/mhPINS cells was also detected.    Results  HepG2/mhPINS-cells grew well. The target fragment of 286 bp was acquired in RT-PCR. A special strap of 34×10

3

was exhibited in Western blot. Insulin and C peptide were detected in the culture supernatant of HepG2/mhPINS cells by radioimmunoassay. The results of HepG2/pLXSN cells in the detections above were all negative. As shown in detection of glucose-stimulating insulin secretion in HepG2/mhPINS cells

there were no significant differences in the insulin secretion at 0.1

10

20 mmol/L glucose concentration (P0.05).    Conclusion  HepG2 cells expressing mhPINS gene were successfully constructed

and the mature insulin secretion was made in HepG2/mhPINS cells.