Long non-coding RNA small nucleolar RNA host gene 1 alleviates the progression of epilepsy by regulating the miR-181a/BCL-2 axis in vitro

Life Sci. 2021 Feb 15:267:118935. doi: 10.1016/j.lfs.2020.118935. Epub 2020 Dec 31.

Abstract

Purpose: Long non-coding RNAs (lncRNAs) have been reported to be involved in regulating epilepsy. The purpose of this study is to investigate the possibly regulatory mechanism of small nucleolar RNA host gene 1 (SNHG1) on epilepsy.

Methods: Quantitative real-time PCR was utilized to detect the expression of SNHG1, microRNA (miR)-181a, and B-cell lymphoma-2 (BCL-2). Through an enzyme-linked immunosorbent assay, the levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, and cyclooxygenase-2 (COX-2) were determined. The viability and apoptosis of CTX-TNA2 cells were measured using MTT assay and flow cytometry analysis, respectively. Western blot assay was performed to analyze the protein levels of Bcl-2, BCL2-associated X, and Caspase-3. The relationships between miR-181a and SNHG1/BCL-2 were confirmed by the dual-luciferase reporter assay.

Results: SNHG1 expression was down-regulated in EP tissues and kainic acid (KA)-induced CTX-TNA2 cells. The apoptosis and release of inflammatory factors (TNF-α, IL-1β, IL-6, and COX-2) in KA-induced CTX-TNA2 cells were suppressed by SNHG1 overexpression and promoted by miR-181a up-regulation. In addition, we confirmed that SNHG1 targeted miR-181a, whereas BCL-2 was a target gene of miR-181a. Negative correlations between SNHG1 and miR-181a, as well as miR-181a and BCL-2 were exhibited. Both the up-regulation of miR-181a and down-regulation of BCL-2 reversed the inhibiting effects of SNHG1 on apoptosis and inflammatory response of KA-induced CTX-TNA2 cells, and the promoting effect upon cell viability.

Conclusions: SNHG1 alleviated the progression of EP by modulating the miR-181a/BCL-2 axis in vitro, thus SNHG1 could act as a possible therapeutic target for treating EP.

Keywords: BCL-2; Epilepsy; lncRNA SNHG1; miR-181a.

MeSH terms

  • Animals
  • Apoptosis / physiology
  • Cell Survival / physiology
  • Disease Models, Animal
  • Disease Progression
  • Epilepsy / genetics*
  • Epilepsy / metabolism
  • Interleukin-1beta / metabolism
  • Male
  • MicroRNAs / genetics*
  • MicroRNAs / metabolism
  • Proto-Oncogene Proteins c-bcl-2 / genetics
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • RNA, Long Noncoding / genetics*
  • RNA, Long Noncoding / metabolism
  • RNA, Small Nucleolar / pharmacology
  • Rats
  • Rats, Sprague-Dawley
  • Signal Transduction
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Interleukin-1beta
  • MIRN181 microRNA, rat
  • MicroRNAs
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Long Noncoding
  • RNA, Small Nucleolar
  • Tumor Necrosis Factor-alpha