Wilms tumor 1 regulates lipid accumulation in human endometrial stromal cells during decidualization
Wilms tumor 1 regulates lipid accumulation in human endometrial stromal cells during decidualization
复制标题
DOI：
10.1074/jbc.ra120.012841
复制
发表时间：
2020-02
期刊：
影响因子：
--
通讯作者：
I. Tamura;Haruka Takagi;Yumiko Doi-Tanaka;Yuichiro Shirafuta;Yumiko Mihara;Masahiro Shinagawa;R. Maekawa;T. Taketani;Shun Sato;H. Tamura;N. Sugino
中科院分区：
文献类型：
--
作者：
I. Tamura;Haruka Takagi;Yumiko Doi-Tanaka;Yuichiro Shirafuta;Yumiko Mihara;Masahiro Shinagawa;R. Maekawa;T. Taketani;Shun Sato;H. Tamura;N. Sugino
我们之前报道过转录因子Wilms瘤1（WT1）在人子宫内膜基质细胞（ESC）蜕膜化过程中调节胰岛素样生长因子结合蛋白1（IGFBP-1）和催乳素（PRL）的表达。然而，WT1 在蜕膜化中的其他作用仍有待完全阐明。在这里，我们研究了 WT1 在蜕膜化过程中如何调节人类 ESC 的生理功能。我们将从增殖期子宫内膜分离的 ESC 与 cAMP 一起孵育以诱导蜕膜化，用 siRNA 敲低 WT1，并产生三种类型的处理细胞（未处理细胞、cAMP 处理细胞和 cAMP 处理 + WT1 敲除细胞）。为了识别 WT1 调控的基因，我们使用基因微阵列并比较了这三种治疗方法中获得的转录组数据。我们观察到 WT1 在蜕膜化过程中上调 121 个基因，其中包括几个参与脂质转运的基因。 WT1 敲低抑制了 cAMP 诱导的 ESC 中的脂质积累 (LA)。为了研究 WT1 调节 LA 的机制，我们重点关注参与脂蛋白摄取的极低密度脂蛋白受体 (VLDLR)。我们发现 cAMP 上调 VLDLR，而 WT1 敲低则抑制它。 ChIP 检测结果显示，cAMP 增加了 WT1 向 VLDLR 基因启动子区域的募集，表明 WT1 调节 VLDLR 表达。此外，VLDLR 敲低抑制了 cAMP 诱导的 LA，而 VLDLR 过表达则恢复了 WT1 敲低引起的 LA 抑制。综上所述，我们的结果表明，WT1 通过在蜕膜化过程中上调人 ESC 中 VLDLR 的表达来增强脂质储存。
We previously reported that the transcription factor Wilms tumor 1 (WT1) regulates the expression of insulin-like growth factor-binding protein-1 (IGFBP-1) and prolactin (PRL) during decidualization of human endometrial stromal cells (ESCs). However, other roles of WT1 in decidualization remain to be fully clarified. Here, we investigated how WT1 regulates the physiological functions of human ESCs during decidualization. We incubated ESCs isolated from proliferative-phase endometrium with cAMP to induce decidualization, knocked down WT1 with siRNA, and generated three types of treatments (nontreated cells, cAMP-treated cells, and cAMP-treated + WT1-knockdown cells). To identify WT1-regulated genes, we used gene microarrays and compared the transcriptome data obtained among these three treatments. We observed that WT1 up-regulates 121 genes during decidualization, including several genes involved in lipid transport. The WT1 knockdown inhibited lipid accumulation (LA) in the cAMP-induced ESCs. To examine the mechanisms by which WT1 regulates LA, we focused on very low-density lipoprotein receptor (VLDLR), which is involved in lipoprotein uptake. We found that cAMP up-regulates VLDLR and that the WT1 knockdown inhibits it. Results of ChIP assays revealed that cAMP increases the recruitment of WT1 to the promoter region of the VLDLR gene, indicating that WT1 regulates VLDLR expression. Moreover, VLDLR knockdown inhibited cAMP-induced LA, and VLDLR overexpression reverted the suppression of LA caused by the WT1 knockdown. Taken together, our results indicate that WT1 enhances lipid storage by up-regulating VLDLR expression in human ESCs during decidualization.