Background:Systemic lupus erythematosus(SLE)is a complex autoimmune disease,and the mechanism of SLE is yet to be fully elucidated.The aim of this study was to explore the role of two-pore segment channel 2(TPCN2)in S...Background:Systemic lupus erythematosus(SLE)is a complex autoimmune disease,and the mechanism of SLE is yet to be fully elucidated.The aim of this study was to explore the role of two-pore segment channel 2(TPCN2)in SLE pathogenesis.Methods:Quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to detect the expression ofTPCN2 in SLE.We performed a loss-of-function assay by lentiviral construct in Jurkat and THP-1 cell.Knockdown ofTPCN2 were confirmed at the RNA level by qRT-PCR and protein level by Western blotting.Cell Count Kit-8 and flow cytometry were used to analyze the cell proliferation,apoptosis,and cell cycle ofTPCN2-deficient cells.In addition,gene expression profile ofTPCN2-deficient cells was analyzed by RNA sequencing(RNA-seq).Results:TPCN2 knockdown with short hairpin RNA(shRNA)-mediated lentiviruses inhibited cell proliferation,and induced apoptosis and cell-cycle arrest of G2/M phase in both Jurkat and THP-1 cells.We analyzed the transcriptome of knockdown-TPCN2-Jurkat cells,and screened the differential genes,which were enriched for the G2/M checkpoint,complement,and interleukin-6-Janus kinase-signal transducer and activator of transcription pathways,as well as changes in levels of forkhead box O,phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin,and T cell receptor pathways;moreover,TPCN2 significantly influenced cellular processes and biological regulation.Conclusion:TPCN2 might be a potential protective factor against SLE.展开更多
基金This work was supported by a grant from the National Natural Science Foundation of China(No.81872516)。
文摘Background:Systemic lupus erythematosus(SLE)is a complex autoimmune disease,and the mechanism of SLE is yet to be fully elucidated.The aim of this study was to explore the role of two-pore segment channel 2(TPCN2)in SLE pathogenesis.Methods:Quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to detect the expression ofTPCN2 in SLE.We performed a loss-of-function assay by lentiviral construct in Jurkat and THP-1 cell.Knockdown ofTPCN2 were confirmed at the RNA level by qRT-PCR and protein level by Western blotting.Cell Count Kit-8 and flow cytometry were used to analyze the cell proliferation,apoptosis,and cell cycle ofTPCN2-deficient cells.In addition,gene expression profile ofTPCN2-deficient cells was analyzed by RNA sequencing(RNA-seq).Results:TPCN2 knockdown with short hairpin RNA(shRNA)-mediated lentiviruses inhibited cell proliferation,and induced apoptosis and cell-cycle arrest of G2/M phase in both Jurkat and THP-1 cells.We analyzed the transcriptome of knockdown-TPCN2-Jurkat cells,and screened the differential genes,which were enriched for the G2/M checkpoint,complement,and interleukin-6-Janus kinase-signal transducer and activator of transcription pathways,as well as changes in levels of forkhead box O,phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin,and T cell receptor pathways;moreover,TPCN2 significantly influenced cellular processes and biological regulation.Conclusion:TPCN2 might be a potential protective factor against SLE.
