Analysis of Foxo1-regulated genes using Foxo1-deficient pancreatic β cells
Analysis of Foxo1-regulated genes using Foxo1-deficient pancreatic β cells
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DOI：
10.1111/j.1365-2443.2012.01625.x
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发表时间：
2012-09-01
期刊：
影响因子：
2.1
通讯作者：
Miyazaki, Jun-ichi
中科院分区：
文献类型：
--
作者：
Miyazaki, Satsuki;Minamida, Rie;Miyazaki, Jun-ichi
一些报告表明，Foxo 1是分化、生长和代谢的关键调节因子，参与胰腺β细胞功能。然而，由于缺乏Foxo 1缺陷的β细胞，详细的分析受到阻碍。为了阐明Foxo 1在β细胞中的功能，我们产生了具有可诱导Foxo 1缺失的β细胞系。我们产生了一个条件性敲除小鼠品系，其中Cre重组酶删除Foxo 1基因。然后，我们建立了一个β细胞系，从胰岛素瘤诱导在这个敲除小鼠的β细胞特异性表达猿猴病毒40 T抗原。在该细胞系（命名为MIN 6-Foxo 1flox/flox）中，腺病毒介导的Cre表达消除了Foxo 1基因，产生了MIN 6-Foxo 1-KO细胞。使用这些敲除和floxed细胞系，我们发现Foxo 1消融增强了高葡萄糖浓度下葡萄糖刺激的胰岛素分泌（GSIS），并增强了β细胞增殖。我们还进行了MIN 6-Foxo 1-KO细胞的DNA微阵列分析感染的腺病毒载体表达组成型活性FOXO 1或控制载体，并确定了几个Foxo 1调节基因，包括一些已知的β细胞功能相关。这些细胞将有助于进一步研究Foxo 1在β细胞中的作用，并可能导致治疗2型糖尿病胰岛素分泌受损的新策略。
Several reports have suggested that Foxo1, a key regulator in differentiation, growth and metabolism, is involved in pancreatic beta-cell function. However, detailed analyses have been hampered by a lack of Foxo1-deficient beta cells. To elucidate Foxo1's function in beta cells, we produced a beta-cell line with inducible Foxo1 deletion. We generated a conditional knockout mouse line, in which Cre recombinase deletes the Foxo1 gene. We then established a beta-cell line from an insulinoma induced in this knockout mouse by the beta-cell-specific expression of simian virus 40 T antigen. In this cell line, designated MIN6-Foxo1flox/flox, adenovirus-mediated Cre expression ablates the Foxo1 gene, generating MIN6-Foxo1-KO cells. Using these knockout and floxed cell lines, we found that Foxo1 ablation enhanced the glucose-stimulated insulin secretion (GSIS) at high glucose concentrations and enhanced beta-cell proliferation. We also conducted DNA microarray analyses of MIN6-Foxo1-KO cells infected with either an adenovirus vector expressing a constitutively active FOXO1 or a control vector and identified several Foxo1-regulated genes, including some known to be related to beta-cell function. These cells should be useful for further studies on Foxo1's roles in beta-cells and may lead to novel strategies for treating the impaired insulin secretion in type 2 diabetes mellitus.