Long noncoding RNA LERFS negatively regulates rheumatoid synovial aggression and proliferation
Yaoyao Zou, … , Song Guo Zheng, Hanshi Xu
Yaoyao Zou, … , Song Guo Zheng, Hanshi Xu
Published September 10, 2018
Citation Information: J Clin Invest. 2018;128(10):4510-4524. https://doi.org/10.1172/JCI97965.
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Research Article Immunology

Long noncoding RNA LERFS negatively regulates rheumatoid synovial aggression and proliferation

Abstract

Fibroblast-like synoviocytes (FLSs) are critical to synovial aggression and joint destruction in rheumatoid arthritis (RA). The role of long noncoding RNAs (lncRNAs) in RA is largely unknown. Here, we identified a lncRNA, LERFS (lowly expressed in rheumatoid fibroblast-like synoviocytes), that negatively regulates the migration, invasion, and proliferation of FLSs through interaction with heterogeneous nuclear ribonucleoprotein Q (hnRNP Q). Under healthy conditions, by binding to the mRNA of RhoA, Rac1, and CDC42 — the small GTPase proteins that control the motility and proliferation of FLSs — the LERFS–hnRNP Q complex decreased the stability or translation of target mRNAs and downregulated their protein levels. But in RA FLSs, decreased LERFS levels induced a reduction of the LERFS–hnRNP Q complex, which reduced the binding of hnRNP Q to target mRNA and therefore increased the stability or translation of target mRNA. These findings suggest that a decrease in synovial LERFS may contribute to synovial aggression and joint destruction in RA and that targeting the lncRNA LERFS may have therapeutic potential in patients with RA.

Authors

Yaoyao Zou, Siqi Xu, Youjun Xiao, Qian Qiu, Maohua Shi, Jingnan Wang, Liuqin Liang, Zhongping Zhan, Xiuyan Yang, Nancy Olsen, Song Guo Zheng, Hanshi Xu

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Figure 3

Effect of LERFS overexpression on the proliferation and apoptosis of RA FLSs.

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Effect of LERFS overexpression on the proliferation and apoptosis of RA ...
(A and B) An EdU incorporation assay was performed to evaluate cell proliferation. Representative images show proliferation of RA FLSs (A) and HC FLSs (C) labeled with EdU (red) and nuclei stained with Hoechst 33342 (blue) (original magnification, ×200). Graphs in A and C indicate the mean ± SEM of 5 independent experiments involving 5 different RA patients or HCs. (B) Detection of cell growth rates in vitro using an MTT assay at the indicated time points after lentivirus infection (D0 indicates the day of infection). Values are expressed relative to D0 as the mean ± SEM of 5 independent experiments. (D and E) Effects of LERFS overexpression on phases of the cell cycle. (D) Representative plots of cell-cycle distribution. (F) LERFS knockdown promotes proliferation by HC FLSs. Representative images are shown (original magnification, ×200). Data are shown as the mean ± SEM of 5 independent experiments involving 5 different RA patients (E) or HCs (F). (G) Effect of LERFS overexpression on apoptosis of RA FLSs. The cellular apoptosis rate was measured by annexin V and 7-AAD staining and detected by flow cytometry. Representative flow plots are shown. Total apoptosis represents the mean ± SEM percentage of 5 independent experiments involving 5 different RA patients. (H) Quantitative measurement of caspase 3/7 activity. Data are expressed relative to vector values and presented as the mean ± SEM of 5 independent experiments involving 5 different RA patients. (I) Effect of LERFS overexpression on FasL-induced apoptosis of RA FLSs. Cells were stimulated with or without 100 ng/ml recombinant human FasL for 24 hours. Total apoptosis represents the mean ± SEM of 3 independent experiments involving 3 different RA patients. *P < 0.05, **P < 0.01, and ***P < 0.001 versus vector, by Student’s t test.