Induction of cell differentiation activates transcription of the Sarco/Endoplasmic Reticulum calcium-ATPase 3 gene (ATP2A3) in gastric and colon cancer cells
Induction of cell differentiation activates transcription of the Sarco/Endoplasmic Reticulum calcium-ATPase 3 gene (ATP2A3) in gastric and colon cancer cells
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DOI：
10.1002/mc.22529
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发表时间：
2017-02-01
影响因子：
4.6
通讯作者：
Zarain-Herzberg, Angel
中科院分区：
文献类型：
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作者：
Flores-Peredo, Lucia;Rodriguez, Gabriela;Zarain-Herzberg, Angel
Sarco/内质网Ca2+- atp酶（SERCAs）将Ca2+泵入内质网腔，调节胞质Ca2+浓度，调节包括细胞生长在内的各种细胞过程。先前的研究报道了胃癌和结肠癌细胞系中SERCA3蛋白表达下调，并表明体外细胞分化使其表达增加。然而，对上皮癌细胞中调节SERCA3表达的转录机制和转录因子知之甚少。在这项工作中，我们证明了SERCA3 mRNA在两种上皮癌细胞系，KATO-III和Caco-2中上调高达45倍，分别诱导组蛋白去乙酰化酶抑制剂（HDACi）和细胞融合。为了评估对分化刺激作出反应的转录元件，我们克隆了人类ATP2A3启动子，生成缺失构建体并将其转染到KATO-III细胞中。通过功能分析，在启动子区域的第一个-135bp内定位了基础和分化响应DNA元件。通过定点诱变和dna -蛋白结合实验，我们发现Sp1、Sp3和Klf-4转录因子与ATP2A3近端启动子元件结合并调节基础基因的表达。我们发现这些因子参与了癌细胞分化过程中ATP2A3表达的增加。本研究首次提供了Sp1、Sp3和Klf-4在诱导上皮癌细胞分化过程中转录调节SERCA3表达的证据。(c) 2016 Wiley Periodicals, Inc.；
The Sarco/Endoplasmic Reticulum Ca2+-ATPases (SERCAs), pump Ca2+ into the endoplasmic reticulum lumen modulating cytosolic Ca2+ concentrations to regulate various cellular processes including cell growth. Previous studies have reported a downregulation of SERCA3 protein expression in gastric and colon cancer cell lines and showed that in vitro cell differentiation increases its expression. However, little is known about the transcriptional mechanisms and transcription factors that regulate SERCA3 expression in epithelial cancer cells. In this work, we demonstrate that SERCA3 mRNA is upregulated up to 45-fold in two epithelial cancer cell lines, KATO-III and Caco-2, induced to differentiate with histone deacetylase inhibitors (HDACi) and by cell confluence, respectively. To evaluate the transcriptional elements responding to the differentiation stimuli, we cloned the human ATP2A3 promoter, generated deletion constructs and transfected them into KATO-III cells. Basal and differentiation responsive DNA elements were located by functional analysis within the first -135bp of the promoter region. Using site-directed mutagenesis and DNA-protein binding assays we found that Sp1, Sp3, and Klf-4 transcription factors bind to ATP2A3 proximal promoter elements and regulate basal gene expression. We showed that these factors participated in the increase of ATP2A3 expression during cancer cell differentiation. This study provides evidence for the first time that Sp1, Sp3, and Klf-4 transcriptionally modulate the expression of SERCA3 during induction of epithelial cancer cell differentiation. (c) 2016 Wiley Periodicals, Inc.