Involvement of hepatocyte nuclear factor 3 in endoderm differentiation of embryonic stem cells
Involvement of hepatocyte nuclear factor 3 in endoderm differentiation of embryonic stem cells
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DOI：
10.1128/mcb.17.7.3817
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发表时间：
1997-07-01
影响因子：
5.3
通讯作者：
Benvenisty, N
中科院分区：
文献类型：
--
作者：
LevinsonDushnik, M;Benvenisty, N
肝细胞核因子3(HNF3)家族转录因子在小鼠内胚层分化早期表达，为研究其在小鼠胚胎干细胞发育中的作用，在胚胎干细胞分化前未检测到HNF3α或HNF3β基因转录产物，在ES细胞向类胚体分化的后期(诱导分化后20天)仅检测到低水平的HNF3β基因。为了研究在ES细胞中过表达hnf3α或-beta的后果，我们将这两个基因导入ES细胞，并测定了组织特异性基因在EB分化过程中的表达水平。具体地，我们检测了白蛋白、囊性纤维化跨膜传导调节因子(Cftr)、磷酸烯醇式丙酮酸羧激酶(Peck)、α1-抗胰蛋白酶、跨甲状腺维甲素、zeta-珠蛋白和神经丝68kd的表达，作为不同细胞系的标志，hnf3β的过表达(以及较少程度的hnf3α)诱导了与内胚层分化相关的基因的表达，即cftr和白蛋白的基因，但不诱导与内胚层晚期分化相关的基因的表达。在高表达HNF3的细胞中，HNF1β的表达较高，而HNF1α和HNF4在这些细胞中的表达较弱，因此，HNF3α和β可能参与了ES细胞的早期内胚层分化，并与其他发育因素一起显然需要在体内诱导内胚层分化。
The transcription factors of the hepatocyte nuclear factor 3 (HNF3) family, which are active in the liver, are expressed early during endoderm differentiation, To study their involvement in early murine development, we examined their role in embryonic stem (ES) cells, HNF3 alpha or HNF3 beta mRNA transcripts were not detected in ES cells before differentiation, and only low levels of HNF3 beta mRNA were detected at a late stage of differentiation of ES cells to embryoid bodies (EB) (20 days after induction of differentiation). To examine the consequences of overexpressing HNF3 alpha or -beta in ES cells, we transfected the two genes into these cells and determined the levels of expression of tissue-specific genes during EB differentiation, Specifically, we examined expression of albumin, cystic fibrosis transmembrane conductance regulator (CFTR), phosphoenolpyruvate carboxykinase (PEPCK), alpha 1-antitrypsin, transthyretin, zeta-globin, and neurofilament 68kd as markers for different cell lineages, Overexpression of HNF3 beta (and to a lesser extent of HNF3 alpha) induced the expression of genes associated with endodermal lineage, namely, the genes for CFTR and albumin, but did not induce the expression of genes involved in late endoderm differentiation, such as the genes for PEPCK and alpha 1-antitrypsin, Moreover, expression of HNF1 beta was highly induced in HNF3-overexpressing cells, while expression of HNF1 alpha and HNF4 was only mildly induced in these cells, Therefore, HNF3 alpha and -beta seem to be involved in early endoderm differentiation of ES cells and together with other developmental factors are apparently needed for the induction of the endodermal lineage in vivo.