CDKN2B antisense RNA 1 expression alleviates idiopathic pulmonary fibrosis by functioning as a competing endogenouse RNA through the miR-199a-5p/Sestrin-2 axis

Mei Yang; Egao Yin; Yiheng Xu; Yongjun Liu; Ting Li; Zhaoxing Dong; Wenlin Tai

doi:10.1080/21655979.2022.2044252

CDKN2B antisense RNA 1 expression alleviates idiopathic pulmonary fibrosis by functioning as a competing endogenouse RNA through the miR-199a-5p/Sestrin-2 axis

Bioengineered. 2022 Mar;13(3):7746-7759. doi: 10.1080/21655979.2022.2044252.

Authors

Mei Yang^{1

2}, Egao Yin¹, Yiheng Xu³, Yongjun Liu¹, Ting Li^{4

5}, Zhaoxing Dong^{4

5}, Wenlin Tai³

Affiliations

¹ Department of Respiration, The Sencond Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, China.
² Department of Respiratory and Critical Care, Affiliated Hospital of Yunnan University, Kunming, Yunnan, China.
³ Department of Clinical Laboratory, Yunnan Molecular Diagnostic Center, the Sencond Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, China.
⁴ Department of Respiration, Hwa Mei Hospital, University of Chinese Academy of Sciences, Ningbo, Zhejiang, China.
⁵ Department of Respiration, Ningbo Institute of Life and Health Industry, University of Chinese Academy of Sciences, Ningbo, Zhejiang, China.

Abstract

Idiopathic pulmonary fibrosis (IPF) is an idiopathic interstitial lung disease. At present, the pathogenesis of IPF has not been fully elucidated, which has affected the development of effective treatment methods. Here, we explored the function and potential mechanism of long noncoding RNA (lncRNA) CDKN2B antisense RNA 1 (CDKN2B-AS1) in IPF.Transforming growth factor-β (TGF-β) and bleomycin (BLM) were used to induce IPF in cells and animal models. Real Time quantitative Polymerase Chain Reaction (RT-qPCR) showed the expression of CDKN2B-AS1, miR-199a-5p and Sestrin-2 (SESN2) in cells and tissues. The double luciferase reporter gene assay confirmed the targeting relationship among CDKN2B-AS1, miR-199a-5p, and SESN2. Related protein levels were detected by Western blot combined with Cell Counting Kit-8 (CCK-8), wound healing, and flow cytometry to analyze cell proliferation, migration, and apoptosis. The pathological characteristics of mouse lung tissue were determined by Hematoxylin-eosin (HE) and Masson staining. We found that the expression of CDKN2B-AS1 was decreased in TGF-β-treated cells and BLM-treated mice. Overexpression of CDKN2B-AS1 inhibited cell proliferation and migration, promoted apoptosis, decreased the expression of fibrosis-related proteins and promoted autophagy. In addition, overexpression of CDKN2B-AS1 alleviated pulmonary fibrosis in BLM-treated mice. Mechanistically, CDKN2B-AS1 acts as a miR-199a-5p sponge to regulate SESN2 expression. Our results indicate the importance of the CDKN2B-AS1/miR-199a-5p/SESN2 axis.

Keywords: CDKN2B-AS1; IPF; SESN2; autophagy; miR-199a-5p.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Line, Tumor
Cell Proliferation / genetics
Idiopathic Pulmonary Fibrosis* / genetics
Mice
MicroRNAs* / genetics
MicroRNAs* / metabolism
RNA, Antisense / genetics
RNA, Long Noncoding* / genetics
RNA, Long Noncoding* / metabolism
Transforming Growth Factor beta

Substances

MicroRNAs
RNA, Antisense
RNA, Long Noncoding
Transforming Growth Factor beta

Grants and funding

This study was supported by the Scientific Research Fund project of Yunnan Education Department [2021Y340 to Mei Yang]; Basic research program of Science and Technology Project of Yunnan Science and Technology Department [202101AY070001-286 to Mei Yang]; The General Program of National Natural Science Foundation of China[8217010399 to Zhaoxing Dong]; Basic Research Program of Yunnan Province (Kunming-Medical Joint Special Project) [2019FE001(-166) to Zhaoxing Dong]; Scientific Research Fund of Yunnan Education Department [2019J1255 to Yiheng Xu].