Role of SF-1 and DAX-1 during differentiation of P19 cells by retinoic acid.
Role of SF-1 and DAX-1 during differentiation of P19 cells by retinoic acid.
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SF-1 和 DAX-1 在视黄酸分化 P19 细胞过程中的作用。
DOI：
10.1002/jcp.22866
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发表时间：
2012
影响因子：
5.6
通讯作者：
Soprano,DianneRobert
中科院分区：
文献类型：
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作者：
Teets,BryanW;Soprano,KennethJ;Soprano,DianneRobert
视黄酸（RA）对胚胎发育和细胞分化至关重要。我们实验室之前的研究表明，阻断P19细胞中RA依赖性的前β细胞白血病转录因子（PBX） mRNA和蛋白水平的增加可以阻止内皮层和神经元的分化。微阵列分析发现，P19细胞的剂量敏感性逆转、肾上腺发育不全临界区、X染色体上的1号基因（DAX‐1）和甾体生成因子（SF‐1）受PBX的调控。为了确定DAX‐1和SF‐1在RA依赖性分化中的作用，我们制备了诱导表达FLAG‐DAX‐1或FLAG‐SF‐1的P19细胞。出乎意料的是，DAX‐1的过表达对RA诱导的P19细胞向内胚层细胞或神经元细胞的分化没有影响。然而，SF - 1过表达阻止了RA依赖性的OCT - 4、DAX - 1的缺失以及COUP - TFI、COUP - TFII和ETS - 1 mRNA水平在内胚层和神经元分化的承诺阶段的增加。令人惊讶的是，SF‐1持续表达7天导致OCT‐4蛋白的RA非依赖性缺失和SSEA‐1表达的RA依赖性缺失。尽管缺乏多能性标记，但这些细胞并没有最终分化为内胚层细胞或神经元细胞。相反，这些细胞获得了许多类固醇生成酶的表达，其模式与肾上腺细胞一致。最后，我们发现了一个反馈回路的证据，在这个反馈回路中，PBX降低了SF - 1 mRNA水平，而SF - 1的持续表达阻止了RA依赖性的PBX水平的增加。综上所述，这些数据表明SF - 1在P19细胞分化过程中起着动态作用，并可能在早期胚胎发生过程中发挥作用。j .细胞。中国生物医学工程学报，26(2):1501-1511,2012。©2011 Wiley期刊公司
Retinoic acid (RA) is critical for embryonic development and cellular differentiation. Previous work in our laboratory has shown that blocking the RA‐dependent increase in pre‐β cell leukemia transcription factors (PBX) mRNA and protein levels in P19 cells prevents endodermal and neuronal differentiation. Dosage‐sensitive sex reversal, adrenal hypoplasia critical region, on chromosome X, gene 1 (DAX‐1) and steroidogenic factor (SF‐1) were found by microarray analysis to be regulated by PBX in P19 cells. To determine the roles of DAX‐1 and SF‐1 during RA‐dependent differentiation, P19 cells that inducibly express either FLAG‐DAX‐1 or FLAG‐SF‐1 were prepared. Unexpectedly, overexpression of DAX‐1 had no effect on the RA‐induced differentiation of P19 cells to either endodermal or neuronal cells. However, SF‐1 overexpression prevented the RA‐dependent loss of OCT‐4, DAX‐1 and the increase in COUP‐TFI, COUP‐TFII, and ETS‐1 mRNA levels during the commitment stages of both endodermal and neuronal differentiation. Surprisingly, continued expression of SF‐1 for 7 days caused the RA‐independent loss of OCT‐4 protein and RA‐dependent loss of SSEA‐1 expression. Despite the loss of well‐characterized pluripotency markers, these cells did not terminally differentiate into either endodermal or neuronal cells. Instead, the cells gained the expression of many steroidogenic enzymes with a pattern consistent with adrenal cells. Finally, we found evidence for a feedback loop in which PBX reduces SF‐1 mRNA levels while continued SF‐1 expression blocks the RA‐dependent increase in PBX levels. Taken together, these data demonstrate that SF‐1 plays a dynamic role during the differentiation of P19 cells and potentially during early embryogenesis. J. Cell. Physiol. 227: 1501–1511, 2012. © 2011 Wiley Periodicals, Inc.