Schlafen 12 Interaction with SerpinB12 and Deubiquitylases Drives Human Enterocyte Differentiation
Schlafen 12 Interaction with SerpinB12 and Deubiquitylases Drives Human Enterocyte Differentiation
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DOI：
10.1159/000492019
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发表时间：
2018-01-01
影响因子：
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通讯作者：
Kiupel, Matti
中科院分区：
文献类型：
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作者：
Basson, Marc D.;Wang, Qinggang;Kiupel, Matti
背景/目的：人类肠细胞分化在发育、禁食、适应和减肥手术过程中会发生改变，但其细胞内调控尚不清楚。我们假设Schlafen 12(SLFN12)调节肠细胞分化。方法：应用激光捕获上皮解剖、qRT-PCR和免疫组织化学方法检测SLFN12在正常和禁食人十二指肠黏膜组织中的表达，以及病毒过表达和siRNA示踪人Caco-2和HIEC6肠上皮细胞中的SLFN12途径。结果：空腹人十二指肠粘膜SLFN12mRNA和蛋白表达减少，同时蔗糖酶异麦芽糖酶等肠细胞标志物减少。SLFN12的过表达增加了Caco-2蔗糖酶-异麦芽糖酶启动子的活性、m RNA和蛋白质，而不依赖于细胞的增殖，并激活了SLFN12可能的启动子。SLFN12共沉淀丝蛋白B12(SERPB12)。失活的SLFN12点突变阻止了SERPB12结合和蔗糖酶-异麦芽糖酶的诱导。SERPB12过表达也诱导蔗糖酶-异麦芽糖酶，而降低SERPB12则阻止SLFN12对蔗糖酶-异麦芽糖酶的影响。SLFN12和SERPB12对蔗糖酶-异麦芽糖酶的诱导作用可通过降低UCHL5或USP14而减弱，并可通过同时降低两者而被阻断。SERPB12刺激USP14活性，但不刺激UCHL5活性。SERPB12与USP14共沉淀，而不与UCHL5共沉淀。此外，SLFN12增加了蔗糖酶-异麦芽糖酶-启动子结合转录因子cdx2的蛋白水平，而不改变cdx2 m RNA。这是通过减少UCHL5和USP14来防止的。我们在体外和体内进一步验证了这一途径。SLFN12或SERPB12过表达诱导人非恶性HIEC-6肠上皮细胞蔗糖酶异麦芽糖酶结论：SLFN12通过SERPB12、脱泛素酶和CDX2途径调节人肠细胞分化。该途径可能被用来调控粘膜萎缩、短肠或肥胖症患者的肠道细胞分化。(C)2018年作者(S)由S.Karger AG，巴塞尔出版
Background/Aims: Human enterocytic differentiation is altered during development, fasting, adaptation, and bariatric surgery, but its intracellular control remains unclear. We hypothesized that Schlafen 12 (SLFN12) regulates enterocyte differentiation. Methods: We used laser capture dissection of epithelium, qRT-PCR, and immunohistochemistry to evaluate SLFN12 expression in biopsies of control and fasting human duodenal mucosa, and viral overexpression and siRNA to trace the SLFN12 pathway in human Caco-2 and HIEC6 intestinal epithelial cells. Results: Fasting human duodenal mucosa expressed less SLFN12 mRNA and protein, accompanied by decreases in enterocytic markers like sucrase-isomaltase. SLFN12 overexpression increased Caco-2 sucrase-isomaltase promoter activity, m RNA, and protein independently of proliferation, and activated the SLFN12 putative promoter. SLFN12 coprecipitated Serpin B12 (SERPB12). An inactivating SLFN12 point mutation prevented both SERPB12 binding and sucrase-isomaltase induction. SERPB12 overexpression also induced sucrase-isomaltase, while reducing SERPB12 prevented the SLFN12 effect on sucrase-isomaltase. Sucrase-isomaltase induction by both SLFN12 and SERPB12 was attenuated by reducing UCHL5 or USP14, and blocked by reducing both. SERPB12 stimulated USP14 but not UCHL5 activity. SERPB12 coprecipitated USP14 but not UCHL5. Moreover, SLFN12 increased protein levels of the sucrase-isomaltase-promoter binding transcription factor cdx2 without altering Cdx2 m RNA. This was prevented by reducing UCHL5 and USP14. We further validated this pathway in vitro and in vivo. SLFN12 or SERPB12 overexpression induced sucrase-isomaltase in human non-malignant HIEC-6 enterocytes. Conclusions: SLFN12 regulates human enterocytic differentiation by a pathway involving SERPB12, the deubiquitylases, and Cdx2. This pathway may be targeted to manipulate human enterocytic differentiation in mucosal atrophy, short gut or obesity. (C) 2018 The Author(s) Published by S. Karger AG, Basel