重组人胰岛素基因在大鼠肝癌细胞中的调节表达

Regulated expression of recombinant human insulin gene in rat hepatoma cells

目的:探讨转染含有葡萄糖反应元件(GLRE)的重组人胰岛素基因大鼠肝癌细胞中不同葡萄糖浓度下胰岛素的分泌情况。方法:利用含有重组质粒[PLXSN-(GLRE)3-BP-1MpINS]的逆转录病毒转染大鼠肝癌CBRH7919细胞,筛选出阳性细胞克隆,调整葡萄糖浓度,测定培养液中胰岛素值。结果:转染后38 h,葡萄糖浓度为0,5,15,25 mmol/L,胰岛素分泌量分别为(5.03±0.72),(9.57±0.49),(32.60±1.96),(43.90±2.30)IU/L,各组间差别显著(P<0.05);转染后1个月,在上述葡萄糖浓度下,胰岛素分泌量分别为(3.57±0.21),(5.30±0.20),(16.27±0.87),(23.23±1.12)IU/L,各组间差别显著(P<0.05)。结论:构建的重组人胰岛素基因具有随培养液中葡萄糖浓度升高调节胰岛素分泌的能力。

Objective： To explore the insulin secretion at different glucose concentrations in rat hepatoma cell line （CBRH7919） which was transfected with recombinant human insulin gene containing glucose reaction element （GLRE）. Methods： Retrovirus containing recombinant plasmid [ PLXSN- （GLRE） 3-BP-1MpINS ] was transfected into rat hepatoma cell line （ CBRH7919 ）, and positive cell clone was selected. The insulin secretion at different glucose concentrations was determined. Results： At the glucoec concentration of 0,5,15 and 25 mmol/L, the insulin secretion was 5.03 ± 0.72, 9.57 ± 0.49, 32.60 ± 1.96, and 43.90 ± 2.30 IU/L 38 hours after transfection and 3.57 ±0.21, 5.30±0.20, 16.27±0.87, and 23.23 ±1.12 IU/L 1 month after transfection, respectively. The differences in insulin secretion were significant between any group and each of other 3 groups （P 〈 0. 05 ）. Conclusion： The recombinant human insulin gene could regulate the insulin secretion with the increase of glucose concentration in vitro.