Long non-coding RNA-non-coding RNA activated by DNA damage inhibition suppresses hepatic stellate cell activation via microRNA-495-3p/sphingosine 1-phosphate receptor 3 axis.

Abstract

Hepatic fibrosis is a damage repair response caused by multiple factors. A growing body of research suggests that long non-coding RNAs (lncRNAs) are involved in a wide range of biological processes, and thus regulate disease progression, including hepatic fibrosis. In this study, we investigated the mechanisms of the long non-coding RNA-non-coding RNA activated by DNA damage (NORAD) in modulating hepatic fibrosis development. Platelet-derived growth factor-BB (PDGF-BB) was used to activate LX-2 hepatic stellate cells (HSCs). The expression of NORAD and microRNA (miR)-495-3p was determined by quantitative real-time polymerase chain reaction (qRT-PCR) analysis. The effects of PDGF-BB on LX-2 cell viability, migration, invasion, and apoptosis were evaluated using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide), Transwell, flow cytometry, and Western blot assays. The activation of HSCs was further verified by examining the expression of the typical markers, alpha smooth muscle actin (α-SMA) and collagen I (Col1α1), using qRT-PCR and Western blot assays. StarBase and dual-luciferase reporter assays were used to assess the binding relationship between miR-495-3p and NORAD. The NORAD levels remarkably increased, whereas the miR-495-3p levels decreased, in PDGF-BB-treated LX-2 cells. miR-495-3p was a putative downstream target of NORAD. NORAD silencing played an anti-fibrotic role by targeting miR-495-3p; this was accomplished by hindering PDGF-BB-treated LX-2 cell viability, migration, and invasion, decreasing the levels of α-SMA and Col1α1, and promoting apoptosis. miR-495-3p protected against hepatic fibrosis by inhibiting sphingosine 1-phosphate receptor 3 (S1PR3) expression. In summary, NORAD silencing inhibited hepatic fibrosis by suppressing HSC activation via the miR-495-3p/S1PR3 axis.

• Authors: Lei Zou, Cuifen Shi, Dawei Wang, Juan Cheng, Qi Wang, Lei Wang, Guoya Yang
• Journal: Bioengineered (Bioengineered) Vol. 13 Issue 3 Pg. 6150-6162 (03 2022) ISSN: 2165-5987 [Electronic] United States
• PMID: 35200103 (Publication Type: Journal Article)

Chemical References

• MIRN495 microRNA, human
• MicroRNAs
• RNA, Long Noncoding
• Sphingosine-1-Phosphate Receptors
• Becaplermin

Topics

• Becaplermin (pharmacology)
• Biological Phenomena
• Cell Proliferation
• DNA Damage
• Hepatic Stellate Cells (metabolism, pathology)
• Humans
• Liver Cirrhosis (genetics, pathology)
• MicroRNAs (metabolism)
• RNA, Long Noncoding (metabolism)
• Sphingosine-1-Phosphate Receptors