SOX2 interferes with the function of CDX2 in bile acid-induced gastric intestinal metaplasia
SOX2 interferes with the function of CDX2 in bile acid-induced gastric intestinal metaplasia
复制标题
SOX2 干扰 CDX2 在胆汁酸诱导的胃肠化生中的功能
DOI：
10.1186/s12935-019-0739-8
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发表时间：
2019-01
期刊：
影响因子：
--
通讯作者：
Shi Yongquan
中科院分区：
文献类型：
--
作者：
Yuan Ting;Ni Zhen;Han Chuan;Min Yali;Sun Lina;Liu Caifang;Shi Miao;Lu wenquan;Wang Na;Du Feng;Wu Qiong;Xie Ning;Shi Yongquan
背景肠上皮化生（IM）是胃癌的癌前病变。动物和临床研究都表明，胆汁酸反流和随后的慢性炎症是IM的关键致病因素。以往的研究表明，在胃分化的关键转录因子SOX 2，在IM的发展过程中下调，而CDX 2，关键的精氨酸特异性转录因子显著上调。然而，目前尚不清楚SOX 2的下调是否促进胃IM的出现或仅仅是一种伴随现象。此外，SOX 2下调IM development.MethodsGastric细胞系在一个剂量依赖性的方式与脱氧胆酸（DCA）的潜在机制尚不清楚。采用免疫组织化学和原位杂交技术检测胃组织芯片中CDX 2和miR-21的表达。进行免疫共沉淀和免疫荧光以确定SOX 2和CDX 2的相互作用。荧光素酶报告基因检测CDX 2的转录活性，并证实miR-21与SOX 2 3′-UTR结合。采用蛋白质印迹法检测SOX 2、CDX 2和下游IM特异性基因的蛋白水平。qRT-PCR检测miR-21、SOX 2、CDX 2及下游IM特异性基因的mRNA水平。此外，我们证明SOX 2过表达可显著抑制胆汁酸和外源性CDX 2诱导的IM特异性基因表达，包括KLF 4、钙粘蛋白17和HNF 4 α的表达。相反，SOX 2敲低具有相反的效果。双荧光素酶报告基因检测表明，SOX 2过表达可显著抑制HEK 293 T细胞中CDX 2的转录活性。CDX 2和SOX 2可在细胞核内形成蛋白复合物。胆汁酸诱导miR-21的表达。结论在胆汁酸诱导的肠化过程中，SOX 2可以干扰CDX 2的转录活性，miR-21可能在这一过程中发挥重要作用，这为胃癌的预防提供了新的思路。
BackgroundIntestinal metaplasia (IM) is a premalignant lesion associated with gastric cancer. Both animal and clinical studies have revealed that bile acid reflux and subsequent chronic inflammation are key causal factors of IM. Previous studies indicated that SOX2, the key transcription factor in gastric differentiation, was downregulated during IM development while CDX2, the pivotal intestine-specific transcription factor was upregulated significantly. However, it remains unclear whether the downregulation of SOX2 promotes gastric IM emergence or is merely a concomitant phenomenon. In addition, the underlying mechanisms of SOX2 downregulation during IM development are unclear.MethodsGastric cell lines were treated with deoxycholic acid (DCA) in a dose-dependent manner. The expression of CDX2 and miR-21 in gastric tissue microarray were detected by immunohistochemistry and in situ hybridization. Coimmunoprecipitation and immunofluorescence were performed to ascertain the interaction of SOX2 and CDX2. Luciferase reporter assays were used to detect the transcriptional activity of CDX2, and confirm miR-21 binding to SOX2 3′-UTR. The protein level of SOX2, CDX2 and downstream IM-specific genes were investigated using western blotting. mRNA level of miR-21, SOX2, CDX2 and downstream IM-specific genes were detected by qRT-PCR.ResultsBile acid treatment could suppress SOX2 expression and simultaneously induce expression of CDX2 in gastric cell lines. Furthermore, we demonstrated that SOX2 overexpression could significantly inhibit bile acid- and exogenous CDX2-induced IM-specific gene expression, including KLF4, cadherin 17 and HNF4α expression. In contrast, SOX2 knockdown had the opposite effect. A dual-luciferase reporter assay demonstrated that SOX2 overexpression could significantly suppress CDX2 transcriptional activity in HEK293T cells. CDX2 and SOX2 could form protein complexes in the nucleus. In addition, bile acid induced the expression of miR-21. The inhibition of SOX2 in bile acid-treated gastric cell lines was rescued by miR-21 knockdown.ConclusionsThese findings suggested that SOX2 can interfere with the transcriptional activity of CDX2 in bile acid-induced IM and that miR-21 might play a key role in this process, which shed new lights in the prevention of gastric cancer.
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影响因子：
29.4
作者：
Hinoi, T;Lucas, PC;Fearon, ER
通讯作者：
Fearon, ER
影响因子：
3.8
作者：
Tatsugami, Masana;Ito, Masanori;Chayama, Kazuaki
通讯作者：
Chayama, Kazuaki
影响因子：
11.2
作者：
Shen, Jing;Xiao, Zhangang;Cho, Chi H.
通讯作者：
Cho, Chi H.
影响因子：
8.8
作者：
Otsubo, T.;Akiyama, Y.;Yanagihara, K.;Yuasa, Y.
通讯作者：
Yuasa, Y.
DOI：
10.1016/j.diff.2010.10.002
发表时间：
2011-02
期刊：
Differentiation; research in biological diversity
影响因子：
--
作者：
H. Mutoh;M. Sashikawa;K. Sugano
通讯作者：
H. Mutoh;M. Sashikawa;K. Sugano