Lin28 regulates the expression of neuropeptide Y receptors and oocyte-specific homeobox genes in mouse embryonic stem cells.
Lin28 regulates the expression of neuropeptide Y receptors and oocyte-specific homeobox genes in mouse embryonic stem cells.
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DOI：
10.5653/cerm.2012.39.2.87
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发表时间：
2012-06
期刊：
影响因子：
--
通讯作者：
Lee KA
中科院分区：
文献类型：
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作者：
Park GT;Seo YM;Lee SY;Lee KA
已知LIN28可控制胚胎干细胞的增殖和多能性。本研究的目的是通过RNA干扰和基因芯片分析来确定LIN28在小鼠胚胎干细胞中的下游效应。将对照siRNA和Lin28siRNA(Dharamacon)分别导入小鼠胚胎干细胞。从每种类型的mESC中提取总RNA，进行逆转录-聚合酶链式反应(RT-PCR)分析，证实Lin28基因表达下调。将这些RNA标记并与Affymetrix基因芯片小鼠基因组430 2.0阵列杂交。数据分析用GenPlex 3.0软件完成。用实时荧光定量RT-PCR检测筛选出的基因的表达水平。根据基因芯片的统计分析，在LIN28下调的mESCs中，共有500个基因发生了改变(上调384个，下调116个)。经过差异表达基因筛选，筛选出31个基因作为LIN28下调调控的候选基因。其中，神经肽Y5受体和卵母细胞特异性同源盒5基因在Lin28下调的mESCs中显著上调。我们还发现，通过下调LIN28，神经肽Y受体(NPYR)和卵母细胞特异性同源框(OBox)基因家族表达上调。根据这项研究的结果，我们认为LIN28通过调节效应因子如NpyR和OBox家族来控制mESCs的特性。
Lin28 has been known to control the proliferation and pluripotency of embryonic stem cells. The purpose of this study was to determine the downstream effectors of Lin28 in mouse embryonic stem cells (mESCs) by RNA interference and microarray analysis. The control siRNA and Lin28 siRNA (Dharmacon) were transfected into mESCs. Total RNA was prepared from each type of transfected mESC and subjected to reverse transcription-polymerase chain reaction (RT-PCR) analysis to confirm the downregulation of Lin28. The RNAs were labeled and hybridized with an Affymetrix Gene-Chip Mouse Genome 430 2.0 array. The data analysis was accomplished by GenPlex 3.0 software. The expression levels of selected genes were confirmed by quantitative real-time RT-PCR. According to the statistical analysis of the cDNA microarray, a total of 500 genes were altered in Lin28-downregulated mESCs (up-regulated, 384; down-regulated, 116). After differentially expressed gene filtering, 31 genes were selected as candidate genes regulated by Lin28 downregulation. Among them, neuropeptide Y5 receptor and oocyte-specific homeobox 5 genes were significantly upregulated in Lin28-downregulated mESCs. We also showed that the families of neuropeptide Y receptor (Npyr) and oocyte-specific homeobox (Obox) genes were upregulated by downregulation of Lin28. Based on the results of this study, we suggest that Lin28 controls the characteristics of mESCs through the regulation of effectors such as the Npyr and Obox families.