LINC00837/miR-671-5p/SERPINE2 functional axis promotes pathological processes of fibroblast-like synovial cells in rheumatoid arthritis

doi:10.12122/j.issn.1673-4254.2025.02.18

Abstract

Abstract:

Objective To investigate the regulatory effect of LINC00837/miR-671-5p/SERPINE2 functional axis on pathological processes of fibroblast-like synovial cells (FLS) in rheumatoid arthritis (RA). Methods RA-FLS were transfected with a LINC00837 overexpression plasmid (pcDNA3.1-LINC00837), a LINC00837 interference plasmid (siRNA-LINC00837), or their respective negative control plasmids (pcDNA3.1-NC and siRNA-NC). Dual luciferase was used to verify the targeting relationship between LINC00837 and miR-671-5p and between miR-671-5p and SERPINE2. RT-qPCR was used to detect the expression levels of LINC00837, miR-671-5p and SERPINE2 in normal FLS or the transfected cells, whose proliferation and migration abilities were assessed using Edu assay and scratch healing assay and by detecting the expression levels of Ki-67, PCNA, E-cadherin and N-cadherin with Western blotting. The changes in cellular secretion of the inflammatory cytokines (TNF‑α, IL-17, IL-4 and IL-10) were examined using ELISA. Results Dual luciferase reporter gene assay showed that LINC00837 was capable of binding to the 3'-UTR of miR-671-5p, and the latter bound to the 3-UTR of SERPINE2 at specific binding sites between them. Compared with normal FLS, RA-FLS showed significantly increased expressions of LINC00837 and SERPINE2, lowered miR-671-5p expression and enhanced proliferation and migration abilities with increased expressions of pro-inflammatory cytokines and reduced expressions of anti-inflammatory cytokines. Transfection of RA-FLS with pcDNA-LINC00837 further enhanced cell proliferation and migration and the changes in the inflammatory cytokines, while transfection with si-LINC00837 produced the opposite changes. Conclusion RA-FLS have a LINC00837/miR-671-5p/SERPINE2 functional axis, which regulates cell proliferation, migration and secretion of inflammatory factors, and interventions targeting LINC00837 may provide a potential strategy to regulate the pathological processes in RA-FLS.

Key words: rheumatoid arthritis, fibroblast-like synovial cells, LINC00837, proliferation, migration, inflammatory factors

Zhoufang CAO, Yuan WANG, Mengna WANG, Yue SUN, Feifei LIU. LINC00837/miR-671-5p/SERPINE2 functional axis promotes pathological processes of fibroblast-like synovial cells in rheumatoid arthritis[J]. Journal of Southern Medical University, 2025, 45(2): 371-378.

Figures/Tables 5

Tab.1 Primer sequences for RT-qPCR of the target genes

Gene	Amplicon size (bp)	Forward primer (5'→3')	Reverse primer (5'→3')
Hu-GAPDH	138	CGGATTTGGTCGTATTGG	GGTGGAATCATATTGGAACA
Hu-U6	94	CTCGCTTCGGCAGCACA	AACGCTTCACGAATTTGCGT
hsa-miR-671-5p		GACTATATAAGCCCTGGAGGGG	AGTGCAGGGTCCGAGGTATT
hsa-miR-671-5p RT		GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACCTCCAG
Hu-Linc00837	139	GCCCGCTTAACTGCTGAAAG	GGACCAACTGCTACAACCCA
Hu-SERPINE2	170	CTCTTTCCGGCTGTGACCCTC	GCCAGTTCATGGTTCCTTCCA

Fig.1 Targeting relationship between LINC00837 and miR-671-5p and between miR-671-5p and SERPINE2. A: Binding site between LINC00837 and miR-671-5p predicted by Targetscan. B: Binding site between miR-671-5p and SERPINE2 predicted by Targetscan. C: Fluorase activity is decreased in miR-671-5p mimic+LINC00837-WT co-transfection group. D: Fluorase activity is decreased miR-671-5p mimic+SERPINE2-WT co-transfection group. **P<0.01.

Fig.2 Expression of LINC00837, miR-671-5p and SERPINE2 in normal FLS (control) and RA-FLS after transfection. **P<0.01, ##P<0.01.

Fig.3 Effects of LINC00837 overexpression or knockdown on proliferation and migration abilities of RA-FLS. A, C: Cell proliferation detected by Edu assay (Original magnification: ×200); B, D: Cell migration detected by scratch healing assay (×100). **P<0.01,##P<0.01.

Fig.5 Effects of LINC00837 overexpression or knockdown on expressions of inflammatory factors TNF-α, IL-17, IL-4 and IL-10 in RA-FLS. **P<0.01, ##P<0.01.

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