LncRNA NEAT1 regulated diabetic retinal epithelial-mesenchymal transition through regulating miR-204/SOX4 axis

Yang Yang; Jing Zhou; Wei Hong Li; Zhi Xiong Zhou; Xiao Bo Xia

doi:10.7717/peerj.11817

LncRNA NEAT1 regulated diabetic retinal epithelial-mesenchymal transition through regulating miR-204/SOX4 axis

PeerJ. 2021 Jul 23:9:e11817. doi: 10.7717/peerj.11817. eCollection 2021.

Authors

Yang Yang^{1

2}, Jing Zhou², Wei Hong Li², Zhi Xiong Zhou², Xiao Bo Xia^{1

3}

Affiliations

¹ Eye center of Xiangya Hospital, Central South University, Changsha, Hunan Province, China.
² Department of Ophthalmology, the First People's Hospital of Yueyang, Yueyang, Hunan, China.
³ Hunan Key Laboratory of Ophthalmology, Central South University, Chang sha, Hunan, China.

Abstract

Aim: Epithelial-mesenchymal transition (EMT) of retinal pigment epithelium (RPE) cells is the key of the development of diabetic retinopathy (DR), and lncRNA NEAT1 could accelerate EMT in diabetic nephropathy. Meanwhile, as a diabetes susceptibility gene, whether sex-determining region Y-related (SRY) high-mobility group box 4 (SOX4) has relationship with lncRNA NEAT1 in DR remains unclear.

Methods: Firstly, NEAT1, SOX4 and miR-204 were evaluated by qRT-PCR (quantitative reverse-transcriptase PCR) under high glucose condition. Then, cell viability, proliferation, migration and invasion were respectively detected by MTT, BrdU staining, wound healing and transwell assay after NEAT1 knockdown or miR-204 overexpression. Also, the EMT-related proteins were examined by western blot and cell immunofluorescence assay. In order to confirm the relationship between miR-204 and NEAT1 or SOX4, dual luciferase reporter gene assay was conducted. At the same time, the protein levels of SOX4 and EMT-related proteins were investigated by immunohistochemistry in vivo.

Results: High glucose upregulated NEAT1 and SOX4 and downregulated miR-204 in ARPE19 cells. NEAT1 knockdown or miR-204 overexpression inhibited the proliferation and EMT progression of ARPE19 cells induced by high glucose. NEAT1 was identified as a molecular sponge of miR-204 to increase the level of SOX4. The effect of NEAT1 knockdown on the progression of EMT under high glucose condition in ARPE19 cells could be reversed by miR-204 inhibitor. Also, NEAT1 knockdown inhibited retinal EMT in diabetic mice.

Conclusion: NEAT1 regulated the development of EMT in DR through miR-204/SOX4 pathway, which could provide reference for clinical prevention and treatment.

Keywords: Epithelial-mesenchymal transition; NEAT1; SOX4; miR-204; Diabetic retinopathy.

Grants and funding

This work was supported by the National Natural Science Foundation of China (NSFC) Grants 81670858 (to Xiaobo Xia). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.