Expression and significance of long noncoding RNA nuclear-enriched abundant transcript 1 (lncRNA NEAT1) in pregnant women with systemic lupus erythematosus
10.3760/cma.j.cn112309-20190823-00270
- VernacularTitle:长链非编码核富含丰富的转录本1(lncRNA NEAT1)在妊娠合并系统性红斑狼疮患者中的表达及意义
- Author:
Fei LIANG
1
;
Yahui XU
;
Rui CHEN
;
Jun YAN
;
Yu WANG
Author Information
1. 河南省人民医院妇产科，郑州 450000
- From:
Chinese Journal of Microbiology and Immunology
2020;40(9):684-689
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the function and mechanism of long noncoding RNA nuclear-enriched abundant transcript 1 (lncRNA NEAT1)-mediated epigenetic regulation of Th2 cell differentiation and development in pregnant women with systemic lupus erythematosus (SLE).Methods:This study involved 11 women with normal singleton pregnancy (control group) and 15 pregnant women with SLE who delivered in the Henan Provincial People′s Hospital from July 1, 2014 to July 1, 2019. Peripheral blood mononuclear cells (PBMCs) were collected and analyzed by qPCR to detect the expression of NEAT1 at mRNA level. ELISA and flow cytometry were used to detect the expression of IFN-γ and IL-4 at protein level. Na?ve CD4 + T cells were sorted out by flow cytometry. RNA binding protein immunoprecipitation (RIP) was performed to detect the binding of EZH2 to NEAT1. After knockdown of NEAT1 expression, Real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to detect the expression of itchy E3 ubiguitin protein ligase(ITCH) at mRNA and protein levels. Chromatin immunoprecipitation (ChIP) was used to detect the abundance of EZH2 at ITCH promoter in pregnant patients with SLE. ELISA was used to detect IL-4 level after overexpression of NEAT1 and ITCH. Statistical data analysis was performed with t test. Results:The expression of NEAT1 at mRNA level in peripheral blood of pregnant women with SLE was significantly higher than that in controls. IFN-γ levels were significantly reduced, while IL-4 levels were significantly increased in pregnant women with SLE than in controls. RIP analysis revealed that there was a great enrichment of NEAT1 in the na?ve CD4 + T cells using anti-EZH2 compared to the control group. After knocking down the expression of NEAT1, the mRNA and protein levels of ITCH were significantly increased. ChIP assay demonstrated that EZH2 was recruited to the promoter of ITCH in pregnant women with SLE. ITCH significantly inhibited the production of IL-4 by na?ve CD4 + T cells, while overexpression of NEAT1 upregulated the expression of IL-4 at protein level. Conclusions:LncRNA NEAT1 was significantly up-regulated in pregnant women with SLE. It recruited EZH2 to the promoter of ITCH and promoted the differentiation of na?ve CD4 + T cells to Th2 cells, resulting Th1/Th2 imbalance and affecting disease progression.
