Noncoding Transcribed Ultraconserved Region (T-UCR) uc.261 Participates in Intestinal Mucosa Barrier Damage in Crohn's Disease
Noncoding Transcribed Ultraconserved Region (T-UCR) uc.261 Participates in Intestinal Mucosa Barrier Damage in Crohn's Disease
复制标题
非编码转录超保守区 (T-UCR) uc.261 参与克罗恩病的肠粘膜屏障损伤。
DOI：
10.1097/mib.0000000000000945
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发表时间：
2016-12-01
影响因子：
4.9
通讯作者：
Ran, Zhi Hua
中科院分区：
文献类型：
--
作者：
Qian, Xiao Xian;Peng, Jiang Chen;Ran, Zhi Hua
背景：迄今为止，人类基因组中已发现 481 个超保守区域（UCR）。我们的目的是研究克罗恩病 (CD) 中转录的 UCR (T-UCR) 特征，并确定 T-UCR 是否是 uc。 261 人参与了肠粘膜屏障损伤。方法：使用 Arraystar 人类 T-UCR 微阵列在活动性 CD 粘膜中筛选 T-UCR，并通过定量实时逆转录 PCR 以及紧密连接蛋白 (TJP) 进行验证，包括连接粘附分子-A、occludin、claudin-1 和 zonula occlusionn-1。通过 RNA 荧光原位杂交验证了活性 CD 粘膜中的 T-UCR uc.261。 Caco2 和 T84 细胞用于测定跨上皮电阻。通过定量实时逆转录 PCR 和蛋白质印迹评估 Cdc42、蛋白激酶 C zeta、PAR3 和 PAR6。采用细胞免疫荧光法检测TJP的组装情况。结果：4个T-UCR显着上调（uc.290-、uc.144-、uc.261-和uc.477+），4个T-UCR显着下调（uc.166-、uc.141-、uc.478+和uc.479+）。 Uc.261 与紧密连接形成过程中的跨上皮电阻呈负相关。活动性 CD 粘膜中 TJP 的水平降低。大多数uc.261位于结肠上皮细胞的细胞质中。 uc.261 的过度表达可降低跨上皮电阻、抑制 TJP 的表达和组装、激活 Cdc42 并抑制蛋白激酶 C zeta。 TNF-α处理的细胞中uc.261的沉默可逆转紧密连接损伤。结论：uc.261的过度表达参与肠粘膜屏障损伤。抑制 uc.261 可逆转炎症中紧密连接的损伤。减弱 uc.261 过度表达可能是治疗 CD 患者的合理策略。
Background: To date, 481 ultraconserved regions (UCRs) have been discovered in human genome. We aimed to investigate the transcribed UCR (T-UCR) characteristics in Crohn's disease (CD) and determine whether T-UCR uc. 261 participated in intestinal mucosa barrier damage.Methods: T-UCRs were screened in active CD mucosa using the Arraystar Human T-UCR Microarray and validated with quantitative real-time reverse transcription PCR, together with tight junction proteins (TJPs) including junctional adhesion molecule-A, occludin, claudin-1, and zonula occluden-1. T-UCR uc.261 in active CD mucosa was validated by RNA fluorescence in situ hybridization. Caco2 and T84 cells were employed to determine transepithelial electrical resistance. Cdc42, protein kinase C zeta, PAR3, and PAR6 were assessed with quantitative real-time reverse transcription PCR and Western blotting. The assembly of TJPs was detected using cell immunofluorescence assay.Results: Four T-UCRs were significantly upregulated (uc.290-, uc.144-, uc.261-, and uc.477+) and 4 T-UCRs were downregulated (uc.166-, uc.141-, uc.478+, and uc.479+). Uc.261 was inversely correlated with transepithelial electrical resistance during tight junction formation. The levels of TJPs were diminished in active CD mucosa. Most uc.261s were located in the cytoplasm of colonic epithelial cells. Overexpression of uc.261 reduced transepithelial electrical resistance, inhibited the expression and assembly of TJPs, activated Cdc42, and suppressed protein kinase C zeta. Silencing of uc.261 in TNF-alpha-treated cells reversed the tight junction damage.Conclusions: Overexpression of uc.261 participates in intestinal mucosa barrier damage. Suppression of uc.261 reverses the damage to tight junction in inflammation. Attenuation of uc.261 overexpression might be a rational strategy to manage patients with CD.