Schlafen 3 Mediates the Differentiating Effects of Cdx2 in Rat IEC-Cdx2L1 Enterocytes.
Schlafen 3 Mediates the Differentiating Effects of Cdx2 in Rat IEC-Cdx2L1 Enterocytes.
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DOI：
10.3109/08941939.2015.1005780
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发表时间：
2015
期刊：
影响因子：
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通讯作者：
Basson MD
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文献类型：
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作者：
Walsh MF;Hermann R;Lee JH;Chaturvedi L;Basson MD
成熟、分化的肠上皮细胞对于正常的肠道功能是必不可少的，并且对于从病理状态中恢复至关重要。人们对调节成人肠道上皮细胞分化的因素知之甚少。参与肠细胞分化的转录因子Cdx 2在成体中仍然表达。由于我们已经在体内和体外的分化中涉及Slfn 3，我们检查了它是否也在分化的IEC-Cdx 2-L1细胞模型中介导分化。在异丙基-β-D-硫代半乳糖苷（IPTG）的控制下，用Cdx 2永久转染IEC Cdx 2-L1细胞，通过暴露于IPTG 16天来刺激其分化。通过定量逆转录-聚合酶链反应从IPTG处理的和对照细胞分离的mRNA的Cdx 2，Slfn 3和绒毛蛋白的转录水平测定。用特异性siRNA降低Slfn 3表达以研究Slfn 3在IPTG分化的细胞中Cdx 2驱动的绒毛蛋白表达中的作用。在IPTG处理的细胞中，Slfn 3和绒毛蛋白的表达显著更高。Slfn 3 siRNA降低了Slfn 3表达并消除了IPTG诱导的绒毛蛋白表达的升高（通过ANOVA，p<0.05）; Cdx 2表达不受Slfn 3 siRNA的影响。数据表明，Cdx 2诱导绒毛蛋白表达并因此诱导分化需要Slfn 3的存在。然而，Slfn 3也必须独立于Cdx 2促进分化，因为通常不表达Cdx 2的IEC-6细胞可以通过多种Slfn 3依赖性机制分化。
Mature, differentiated enterocytes are essential for normal gut function and critical to recovery from pathological conditions. Little is known about the factors that regulate intestinal epithelial cell differentiation in the adult intestine. The transcription factor, Cdx2, involved in enterocytic differentiation, remains expressed in the adult. Since we have implicated Slfn3 in differentiation in vivo and in vitro, we examined whether it also mediated differentiation in the IEC-Cdx2-L1 cell model of differentiation. IEC-Cdx2-L1 cells, permanently transfected with Cdx2 under the control of isopropyl-β-D-thiogalactoside (IPTG), were stimulated to differentiate by 16-day exposure to IPTG. Transcript levels of Cdx2, Slfn 3 and villin were determined by quantitative reverse transcriptase-polymerase chain reaction of mRNA isolated from IPTG-treated and control cells. Slfn3 expression was lowered with specific siRNA to investigate the role of Slfn3 in Cdx2-driven villin expression in IPTG-differentiated cells. Slfn3 and villin expression was significantly greater in IPTG-treated cells. Slfn3 siRNA lowered Slfn3 expression and abolished the IPTG-induced rise in villin expression (p<0.05 by ANOVA); Cdx2 expression was unaffected by Slfn3 siRNA. The data indicate that the presence of Slfn3 is required for Cdx2 to induce villin expression and thus, differentiation. However, Slfn3 must also promote differentiation independently of Cdx2 since IEC-6 cells that do not normally express Cdx2 can be differentiated by a variety of Slfn3-dependent mechanisms.