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...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | zho |
| Published: |
Editorial Department of Journal of Clinical Nephrology
2022-05-01
|
| Series: | Linchuang shenzangbing zazhi |
| Subjects: | |
| Online Access: | http://www.lcszb.com/thesisDetails#10.3969/j.issn.1671-2390.2022.05.008 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850040168670560256 |
|---|---|
| author | Wu Yan-ping Ma Xiao Han Si-ping Wang Xiao-qin |
| author_facet | Wu Yan-ping Ma Xiao Han Si-ping Wang Xiao-qin |
| author_sort | Wu Yan-ping |
| collection | DOAJ |
| description | 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.. |
| format | Article |
| id | doaj-art-93b6e5deb77a429ca099e62fb3ea85e6 |
| institution | DOAJ |
| issn | 1671-2390 |
| language | zho |
| publishDate | 2022-05-01 |
| publisher | Editorial Department of Journal of Clinical Nephrology |
| record_format | Article |
| series | Linchuang shenzangbing zazhi |
| spelling | doaj-art-93b6e5deb77a429ca099e62fb3ea85e62025-08-20T02:56:09ZzhoEditorial Department of Journal of Clinical NephrologyLinchuang shenzangbing zazhi1671-23902022-05-012239640226617956LncRNA XIST promoted the stability of TLR9 mRNA in a FUS-dependent manner to facilitate aerobic glycolysis and proliferation in Th17/CD4+T cellsWu Yan-pingMa XiaoHan Si-pingWang Xiao-qinObjectiveTo 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..http://www.lcszb.com/thesisDetails#10.3969/j.issn.1671-2390.2022.05.008Th17 cellsProliferationlncRNA XISTRNA binding proteinGlomerulonephritis，IgA |
| spellingShingle | Wu Yan-ping Ma Xiao Han Si-ping Wang Xiao-qin LncRNA XIST promoted the stability of TLR9 mRNA in a FUS-dependent manner to facilitate aerobic glycolysis and proliferation in Th17/CD4+T cells Linchuang shenzangbing zazhi Th17 cells Proliferation lncRNA XIST RNA binding protein Glomerulonephritis，IgA |
| title | LncRNA XIST promoted the stability of TLR9 mRNA in a FUS-dependent manner to facilitate aerobic glycolysis and proliferation in Th17/CD4+T cells |
| title_full | LncRNA XIST promoted the stability of TLR9 mRNA in a FUS-dependent manner to facilitate aerobic glycolysis and proliferation in Th17/CD4+T cells |
| title_fullStr | LncRNA XIST promoted the stability of TLR9 mRNA in a FUS-dependent manner to facilitate aerobic glycolysis and proliferation in Th17/CD4+T cells |
| title_full_unstemmed | LncRNA XIST promoted the stability of TLR9 mRNA in a FUS-dependent manner to facilitate aerobic glycolysis and proliferation in Th17/CD4+T cells |
| title_short | LncRNA XIST promoted the stability of TLR9 mRNA in a FUS-dependent manner to facilitate aerobic glycolysis and proliferation in Th17/CD4+T cells |
| title_sort | lncrna xist promoted the stability of tlr9 mrna in a fus dependent manner to facilitate aerobic glycolysis and proliferation in th17 cd4 t cells |
| topic | Th17 cells Proliferation lncRNA XIST RNA binding protein Glomerulonephritis，IgA |
| url | http://www.lcszb.com/thesisDetails#10.3969/j.issn.1671-2390.2022.05.008 |
| work_keys_str_mv | AT wuyanping lncrnaxistpromotedthestabilityoftlr9mrnainafusdependentmannertofacilitateaerobicglycolysisandproliferationinth17cd4tcells AT maxiao lncrnaxistpromotedthestabilityoftlr9mrnainafusdependentmannertofacilitateaerobicglycolysisandproliferationinth17cd4tcells AT hansiping lncrnaxistpromotedthestabilityoftlr9mrnainafusdependentmannertofacilitateaerobicglycolysisandproliferationinth17cd4tcells AT wangxiaoqin lncrnaxistpromotedthestabilityoftlr9mrnainafusdependentmannertofacilitateaerobicglycolysisandproliferationinth17cd4tcells |