LncRNA XIST promoted the stability of TLR9 mRNA in a FUS-dependent manner to facilitate aerobic glycolysis and proliferation in Th17/CD4+T cells
ObjectiveTo explore the expression and function of lncRNA XIST in Th17 cells in IgA nephropathy （IgAN） and examine the regulation of lncRNA XIST on TLR9 mRNA expression.MethodsCD4+T cells were isolated from peripheral blood mononuclear cells （PBMCs） and induced to differentiate into Th17 cells.XIST...
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| Main Authors: | , , , |
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| Format: | Article |
| Language: | zho |
| Published: |
Editorial Department of Journal of Clinical Nephrology
2022-05-01
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| Series: | Linchuang shenzangbing zazhi |
| Subjects: | |
| Online Access: | http://www.lcszb.com/thesisDetails#10.3969/j.issn.1671-2390.2022.05.008 |
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| Summary: | ObjectiveTo explore the expression and function of lncRNA XIST in Th17 cells in IgA nephropathy （IgAN） and examine the regulation of lncRNA XIST on TLR9 mRNA expression.MethodsCD4+T cells were isolated from peripheral blood mononuclear cells （PBMCs） and induced to differentiate into Th17 cells.XIST was over-expressed and silenced by plasmid transfection.Proliferation and apoptosis of Th17 cells with different levels of XIST expression were detected by flow cytometry and CCK-8 assay.Levels of XIST，TLR9 mRNA and protein were detected by quantitative reverse transcription-polymerase chain reaction （qRT-PCR） and Western blot.The effect of XIST on TLR9 expression was predicted by luciferase report analysis and TLR9 mRNA half-life detection.Potential RNA-binding proteins interacting with XIST were searched through Starbase database and detected by RNA immunoprecipitation and Western blot.GraphPad8.0 was utilized for statistical analysis and variables were expressed as average±SD.Student T test or one-way ANOVA was employed.ResultsXIST was distributed predominantly in cytoplasm.It hinted that XIST might be involved in post-transcriptional regulation.A down-regulation of lncRNA XIST expression promoted apoptosis and DNA damage and arrested cell proliferation by regulating TLR9.With an over-expression of lncRNA XIST，TLR9 expression spiked and XIST up-regulated the expression of TLR9 by promoting the stability of TLR9 mRNA.As an RNA-binding protein （RBP），FUS interacted with XIST and FUS antibodies could significantly enrich XIST transcripts.A down-regulation of lncRNA XIST expression had no effect on FUS protein expression.It suggested that XIST might affect FUS function rather than expression.It was further confirmed that a down-regulation of XIST significantly reduced FUS-TLR9 mRNA binding and FUS over-expression rescued a down-regulation and degradation of TLR9 mediated by si-XIST.ConclusionlncRNA XIST and TLR9 play a critical role in Th17 cell proliferation in IgAN through a FUS-mediated signaling pathway.This finding provides new therapeutic ideas for IgAN.. |
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| ISSN: | 1671-2390 |