PIK3R3 regulates ZO-1 expression through the NF-kB pathway in inflammatory bowel disease
PIK3R3 regulates ZO-1 expression through the NF-kB pathway in inflammatory bowel disease
复制标题
PIK3R3 通过 NF-kB 通路调节炎症性肠病中 ZO-1 的表达
DOI：
10.1016/j.intimp.2020.106610
复制
发表时间：
2020-08-01
影响因子：
5.6
通讯作者：
Wang, Jing
中科院分区：
文献类型：
--
作者：
Ibrahim, Sidikjan;Zhu, Xu;Wang, Jing
背景和目的：炎症性肠病（IBD）是结肠炎相关癌（CAC）的主要危险因素。我们以前报道过磷酸肌醇-3-激酶调节亚基3（PIK 3R 3）在结直肠癌（CRC）中过表达，但我们不知道PIK 3R 3在IBD中的作用。方法：我们通过免疫组织化学（IHC）和基因表达综合数据库（GEO）分析来研究PIK 3R 3和ZO-1在IBD患者中的差异表达。将Caco-2细胞暴露于不同条件以评估PIK 3R 3和ZO-1的蛋白质水平变化。用PIK 3R 3/siPIK 3R 3转染Caco-2细胞单层以评估跨上皮电阻。通过免疫印迹和免疫荧光评估紧密连接蛋白的完整性。结果：PIK 3R 3在IBD患者中表达增加，并负性调控ZO-1的表达。在体外，PIK 3R 3通过激活NF-kB通路调节ZO-1。在Caco-2细胞中过表达PIK 3R 3可降低跨上皮电阻（TEER），而在siPIK 3R 3细胞中观察到相反的结果。在动物研究中，N15抑制PIK 3R 3有助于改善DSS诱导的肠通透性。用N15处理的小鼠在结肠组织中表现出较少的TJ破坏。结论：PIK 3R 3在临床IBD患者中增加，伴随ZO-1表达的破坏。PIK 3R 3的抑制减弱了小鼠模型中DSS诱导的IBD症状。这些发现表明PIK 3R 3可能是IBD的治疗靶点。
Background and aims: Inflammatory bowel disease (IBD) are the major risk factor for developing colitis associated cancer (CAC). Previously, we have reported that Phosphoinositide-3-kinase regulatory subunit 3 (PIK3R3) was overexpressed in colorectal cancer (CRC), but we don't know the role of PIK3R3 in IBD.Methods: We investigated the differential expression of PIK3R3 and ZO-1 in IBD patients by using Immunohistochemical (IHC) and Gene Expression Omnibus (GEO) database analysis. Caco-2 cells were exposed to different conditions to assess protein level changes of PIK3R3 and ZO-1. Caco-2 cell monolayers were transfected with PIK3R3/siPIK3R3 to assess transepithelial electrical resistance. Tight junction protein integrity was assessed by immunoblot and immunofluorescence. For further, intestinal permeability and tight junction protein integrity were assessed in animal study to assess the treatment role of PIK3R3 specific inhibitor TAT-N 15 (N15).Results: PIK3R3 was increased in IBD patients, and negatively controlled the expression of ZO-1. In vitro, PIK3R3 regulates ZO-1 by activating NF-kB pathway. Overexpression of PIK3R3 in Caco-2 cells decreased transepithelial electrical resistance (TEER), an opposite result was observed in siPIK3R3 cells. In animal study, inhibition of PIK3R3 by N15 contributed to amelioration of DSS-induced intestinal permeability. Mice treated with N15 exhibited less disruption of TJs in colon tissues.Conclusions: PIK3R3 was increased in clinical IBD patients with accompanying disruption of ZO-1 expression. Inhibition of PIK3R3 attenuated DSS-induced IBD symptoms in a mouse model. These findings indicated that PIK3R3 could be a therapeutic target for IBD.