CV-1细胞中固醇调节元件结合蛋白-1c对Ⅰ型葡萄糖载体mRNA稳定性及蛋白表达的影响

Effects of sterol regulatory element binding protein-1c on stability of glucose transporter Ⅰ mRNA and it's protein expression in CV-1 cells

[目的]探讨在CV-1细胞中固醇调节元件结合蛋白-1c(SREBP-1c)对Ⅰ型葡萄糖载体mRNA稳定性和蛋白表达的影响.[方法]利用DNA克隆技术,将SREBP-1c和其阴性表达体(ADDN)DNA序列重组到表达载体pSV-sport上.用SREBP-1c转染CV-1细胞,应用Northern Blot方法观察SREBP-1c对Ⅰ型葡萄糖载体mRNA的表达及其稳定性的影响,应用Western Blot方法观察SREBP-1c对Ⅰ型葡萄糖载体蛋白表达的影响.[结果]SREBP-1c可增加Ⅰ型葡萄糖载体的mRNA表达水平及稳定性,可提高Ⅰ型葡萄糖载体的蛋白表达水平.[结论]SREBP-1c可增强Ⅰ型葡萄糖载体的蛋白表达,可能通过调节Ⅰ型葡萄糖载体的表达水平而参与胰岛素对血糖的调节过程.

OBJECTIVE To understand the effective of sterol regulatory element binding protein-1c（SREBP-1c） on stability of glucose transporter Ⅰ（GLUT1） mRNA and it＇s protein expression in CV-1 cells.METHODS SREBP-1c cDNA sequence and it＇s negative form（ADDN） were cloned into pSV-sport expression vector.The recombinant SREBP-1c and ADDN were transfected into CV-1 cells,and the stability of GLUT1 mRNA expression and protein expression were detected by time dependent and dose dependent with Northern Blot and Western Blot.RESULTS SREBP-1c stabilizeed the GLUT1 mRNA and increased the GLUT1 protein expression.CONCLUSION SREBP-1c increases GLUT1 expression,and it maybe participate in regulating process of blood glucose level through regulating GLUT1 expression controlled by insulin.