Abstract
AIM: To construct an artificial beta which exhibit glucose-stimulated insulin secretion from a human hepatoma cell line. METHODS: HepG2 cells were infected with recombinant retrovirus carrying glucokinase (gk) gene and mutant pro- insulin (mpin) gene, then selectively cultured with G418 to obtain the positive clones. gk gene and mpin gene tran- scription and expression were identified by radio-immunity, SDS-PAGE, Western-blot and RT-PCR techniques. At last, we tested the dose-response effect of glucose on insulin secretion from the artificial beta cell, and HepG2 cells with mpin gene transferred as control. RESULTS: HepG2 cells with gk gene and mpin gene trans-
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