Affiliation:
1. Department of Clinical Skills Experiment Center The Third Affiliated Hospital of Qiqihar Medical University Qiqihar China
2. Department of Endoscopy Center The Third Affiliated Hospital of Qiqihar Medical University Qiqihar China
3. Department of Gastroenterology The Third Affiliated Hospital of Qiqihar Medical University Qiqihar China
Abstract
AbstractBackgroundLong noncoding RNAs (lncRNAs) and microRNAs (miRNAs) have been reported to play regulatory roles in ulcerative colitis (UC). In this study, we aimed to determine the specific roles and action mechanism of the nuclear paraspeckle assembly transcript 1 (NEAT1) in UC.MethodsReverse transcription‐quantitative polymerase chain reaction (RT‐qPCR) was used to determine the lncRNA NEAT1 and miR‐493‐5p expression levels in patients with UC and healthy volunteers. We determine the forecast linkage points of NEAT1 and miR‐493‐5p using Starbase and those of miR‐493‐5p and Rab27A using TargetScan, and further verified them using a double luciferase gene reporter kit. RT‐qPCR and Western blot analysis were used to determine the lncRNA NEAT1, miR‐493‐5p, and Rab27A expression levels in lipopolysaccharide (LPS)‐induced Caco‐2 cells. Flow cytometry and cell counting kit‐8 were used to assess Caco‐2 cell viability. Tumor necrosis factor‐α, interleukin (IL)‐6, IL‐8, and IL‐1β levels were determined via an enzyme‐linked immunosorbent assay.ResultsExpression levels of NEAT1 were upregulated and those of miR‐493‐5p were downregualted in 10 ng/mL LPS‐treated Caco‐2 cells and patients with UC. Dual‐luciferase gene reporter assay revealed that miR‐493‐5p is linked to NEAT1, and Rab27A is a downstream target of miR‐493‐5p. Overexpression of miR‐493‐5p inhibited the apoptosis and inflammation in LPS‐treated Caco‐2 cells. Moreover, downregulation of lncRNA NEAT1 expression also inhibited the apoptosis and inflammation in LPS‐treated Caco‐2 cells, which was reversed by Rab27A plasmid cotransfection.ConclusionOur results revealed that NEAT1 participates in UC progression by inhibiting miR‐493‐5p expression.
Subject
Immunology,Immunology and Allergy
Cited by
3 articles.
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