• 细胞与分子生物学 •    下一篇

胰岛素刺激对HIT-T15细胞胰岛素原基因表达的影响

张君   

  1. 广西医科大学第一附属医院
  • 收稿日期:2013-09-22 修回日期:2013-12-12 出版日期:2014-04-15 发布日期:2014-04-15
  • 通讯作者: 张君

The Short Term Effect of Insulin on Proinsulin Gene Expression of HIT-T15 Insulinnoma Cells

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  • Received:2013-09-22 Revised:2013-12-12 Published:2014-04-15 Online:2014-04-15
  • Contact: jun ZHANG

摘要:

【摘要】目的 探讨短期外源性胰岛素刺激对HIT-T15胰岛β细胞胰岛素原(PI)基因表达的影响及机制。方法 HIT-T15仓鼠胰岛瘤细胞随机分为4组:含1.4mmol/L葡萄糖完全培养基组作为空白对照(LG组),为抑制内源性胰岛素释放干扰以低糖联合硝苯地平作为条件对照(LGC组),在LGC基础上分别加0.5U/L或5U/L的外源性胰岛素作为实验组(分别为LINS组和HINS组),分别在刺激后0、30、60、90、120min以荧光定量PCR检测各组PI mRNA,免疫组化检测各组细胞胰岛素受体底物1(IRS1)酪氨酸磷酸化水平。结果(1)LINS组、HINS组细胞PI mRNA表达均上调,在刺激后60min PI mRNA表达量达高峰。(2)实验组在刺激后30min IRS1酪氨酸磷酸化水平较对照组明显增高,高、低浓度胰岛素组细胞酪氨酸磷酸化达高峰时间不同。(3)LG组和LGC组间PI mRNA表达及IRS1酪氨酸磷酸化水平无明显差异。结论短期外源性胰岛素能上调胰岛β细胞PI基因的表达,且高浓度的胰岛素较低浓度胰岛素作用强。PI表达调控与IRS1信号传导有关。

关键词: 胰岛素原, HIT-T15细胞, 硝苯地平, 胰岛素受体底物1, 酪氨酸磷酸化

Abstract:

[Abstract] Objective To investigate the short term effect of insulin on proinsulin gene expression of HIT-T15insu? linnoma cells(pancreatic isletβ-cell). Methods The HIT-T15cells were randomly divided into four groups.Blank Con? trol Group (LG) : complete medium contain 1.4mmol/L glucose. Control group (LGC):co-cultured nifedipine with medium in order to restain endogenous insulin release. Experimental group (LINS or HINS) add0.5U/L insulin or5U/L insulin on top of LGC. After being stimulated for0, 30, 60, 90, 120mins, proinsulin (PI) mRNA level were assessed by semi-quantitative RT-PCR. Insulin receptor substrate1(IRS1) tyrosine phosphorylation was detected by immunocytochemistry. Results (1) Expression of PI was up regulated by both LINS and HINS, and peak at60mins. (2) After stimulation for 30mins, the level of IRS1tyrosine phosphorylation in the experimental group was significantly higher than control group, and the peak time be? tween LINS and HINS was different. (3) Between group of LG and LGC, the expression of PI mRNA and IRS1tyrosine phos? phorylation show no difference.Conclusion Short term exogenous insulin stimulation can promote expression of proinsulin genes,which is concentration dependent. The expression and regulation of PI were related with IRS1signal transduction.

Key words: proinsulin, tyrosine phosphorylation