猪Neuronatin基因在pTr2细胞内葡萄糖转运中的功能研究

Study on the Effect of Porcine Neuronatin Gene on Glucose Transport in pTr2 Cells

本研究旨在探索印记基因NNAT 2个转录本NNAT-α、NNAT-β在猪胎盘滋养层细胞(pTr2)中的功能。实验通过NCBI数据库公布的猪NNAT-α、NNAT-β的CDs序列信息设计引物,并构建了pcDNA3.1-NNAT-α和pcDNA3.1-NNAT-β真核表达载体,经过双酶切、连接转化、测序鉴定后,将表达载体转染至pTr2细胞,48 h后通过酶标仪检测细胞内葡萄糖含量的变化,RT-PCR检测NNAT基因、葡萄糖转运基因(GLUT1、GLUT3)和PI3K-AKT通路基因的表达。结果表明在pTr2细胞中过表达NNAT-α、NNAT-β可显著提高细胞内葡萄糖含量、葡萄糖转运基因(GLUT1、GLUT3)和PI3K-AKT通路基因mRNA的表达量。

This experiment was conducted to explore the functions of the two transcripts of the imprinted gene NNAT,NNAT-αand NNAT-β,in porcine placental trophoblast cells(pTr2).In this study,primers were designed based on the CDs sequence of pig NNAT-αand NNAT-βin the NCBI database,and the expression vectors of pcDNA3.1-NNAT-αand pcDNA3.1-NNAT-βwere constructed.After double enzyme digestion and sequencing identification,they were transfected into the pTr2 cells.After transfection for 48 hours,the changes in glucose contents in the cells were detected by a microplate reader,and the expression levels of NNAT gene,glucose transport genes(GLUT1,GLUT3)and PI3K-AKT pathway genes were detected by RT-PCR.The results showed that overexpression of NNAT-αand NNAT-βin pTr2 cells can significantly increase the intracellular glucose content,glucose transport genes(GLUT1,GLUT3)and the genes in the PI3K-AKT pathway.