Downregulation of Long Noncoding RNA Meg3 Affects Insulin Synthesis and Secretion in Mouse Pancreatic Beta Cells
Downregulation of Long Noncoding RNA Meg3 Affects Insulin Synthesis and Secretion in Mouse Pancreatic Beta Cells
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DOI：
10.1002/jcp.25175
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发表时间：
2016-04-01
影响因子：
5.6
通讯作者：
De, Wei
中科院分区：
文献类型：
--
作者：
You, Lianghui;Wang, Ning;De, Wei
越来越多的证据表明长链非编码RNA（lncRNA）参与了多种生物学过程。小鼠母体表达基因3（Meg 3）是一个印记基因，对发育至关重要。在此，我们探讨了Meg 3与小鼠β细胞功能的关系，在体外和体内。Real-time PCR分析显示Meg 3在Balb/c小鼠胰岛中的表达量高于外分泌腺。此外，在T1 DM（NOD雌性小鼠）和T2 DM（db/db小鼠）模型中胰岛中Meg 3的表达降低。葡萄糖在Min 6细胞和分离的小鼠胰岛中动态调节Meg 3表达。通过小干扰RNA（siRNA）敲低小鼠和Min 6细胞中的Meg 3基因，研究Meg 3基因在Min 6细胞中的功能。在体外抑制Meg 3表达后，胰岛素合成和分泌受损，β细胞凋亡率增加。此外，在体内敲低Meg 3导致糖耐量受损和胰岛素分泌减少，与免疫化学测定的胰岛素阳性细胞面积减少一致。值得注意的是，来自Meg 3干扰组的胰岛显示Pdx-1和MafA在mRNA和蛋白水平上的表达显著降低。这些结果表明，Meg 3可能作为一种新的调节剂，通过影响胰岛素产生和细胞凋亡来维持β细胞的身份。(C)2015 Wiley Periodicals，Inc.
Increasing evidence indicates that long noncoding RNAs (lncRNAs) are involved in diverse biological process. Mouse maternal expressed gene 3 (Meg3) is an imprinted gene and essential for development. Here, we explored the relationship between Meg3 and the function of mouse beta cells in vitro and in vivo. Real-time PCR analyses revealed that Meg3 was more abundantly expressed in Balb/cmouse islets than exocrine glands. Moreover, the expression of Meg3 in islets was decreased in T1DM (NOD female mice) and T2DM (db/db mice) models. Meg3 expression was modulated dynamically by glucose in Min6 cells and isolated mouse islets. The function role of Meg3 was investigated in Min6 cells and normal mouse by knockdown of Meg3 using small interfering RNA. After suppression of Meg3 expression in vitro, insulin synthesis and secretion were impaired and the rate of beta cells apoptosis was increased. Moreover, knockdown of Meg3 in vivo led to the impaired glucose tolerance and decreased insulin secretion, consisted with the reduction of insulin positive cells areas by immunochemistry assays. Notably, islets from Meg3 interference groups showed significant decrease of Pdx-1 and MafA expression in mRNA and protein levels. These results indicate that Meg3 may function as a new regulator of maintaining beta cells identity via affecting insulin production and cell apoptosis. (C) 2015 Wiley Periodicals, Inc.