Objectives:Deubiquitinase OTUB2 plays a critical role in the progression of various tumors.However,its specific role in triple-negative breast cancer(TNBC)remains unclear.This study aims to elucidate the biological fu...Objectives:Deubiquitinase OTUB2 plays a critical role in the progression of various tumors.However,its specific role in triple-negative breast cancer(TNBC)remains unclear.This study aims to elucidate the biological function of OTUB2 in TNBC and uncover the underlying mechanisms.Methods:First,we found that the expression of OTUB2 was upregulated in TNBC by bioinformatics analysis,we then validated its expression in TNBC tissues and cells using immunohistochemistry(IHC)and qPCR and plotted the survival curves by Kaplan-Meier method.Gene set enrichment analysis(GSEA)suggested that OTUB2 may be involved in tumor proliferation and metastasis.Further functional assays,including Cell Counting Kit-8(CCK-8),colony formation,Transwell,and wound healing assays,were performed to assess the effects of OTUB2 overexpression and knockdown on TNBC cell proliferation and migration.Additionally,UbiBrowser 2.0 was used to identify OTUB2 substrate proteins and western blotting was conducted to clarify the molecular mechanisms involved.Results:Our results demonstrated that OTUB2 expression was elevated in TNBC and associated with poor prognosis.Overexpression of OTUB2 enhanced the proliferation and migration of TNBC cells,while its knockdown inhibited these processes.Moreover,OTUB2 stabilized tumor necrosis factor receptor-associated factor 6(TRAF6)by deubiquitinating it,leading to activation of the protein kinase B(AKT)pathway.Conclusions:OTUB2 exerts its promoting effects on the progression of TNBC by activating the TRAF6/AKT pathway.展开更多
Objective:To investigate the regulatory effect of miR-146a overexpression on corneal inflammatory response in a mouse model of dry eye,and to analyze its relationship with the IRAK1/TRAF6/NF-кB signaling pathway.Meth...Objective:To investigate the regulatory effect of miR-146a overexpression on corneal inflammatory response in a mouse model of dry eye,and to analyze its relationship with the IRAK1/TRAF6/NF-кB signaling pathway.Methods:A total of 50 SPF-grade BALB/c mice were randomly divided into five groups,with10 mice in each group.Except for the control group,the other four groups were treated with 0.2%benzalkonium chloride(BAC)solution in both eyes to construct a dry eye model.After successful modeling,the control group and model group received NC agomir;the miR antagonist group received miR-146a antagomir;the miR agonist group received miR-146a agomir;and the pathway agonist group received miR-146a agomir+NF-κB activator 2.After four weeks of treatment,the expressions levels of miR-146a,inflammatory factors,and IRAK1/TRAF6/NF-κB signaling pathway proteins were observed and compared among the five groups.Results:After four weeks of treatment,there was a statistically significant difference in the relative expression of miR-146a in the five groups(F=61.058,P<0.001),which was significantly higher in the miR agonist group than in the other four groups.After4 weeks of treatment,there were statistically significant differences in the expression levels of IL-1β,IL-6,IL-8 and TNF-αin the five groups(F=84.757,103.658,55.477,46.762;P<0.001).After four weeks of treatment,there were statistically significant differences in the protein expression levels of IRAK1,TRAF6,NF-κB and IκBαin the five groups(F=62.975,77.173,67.108,29.381;P<0.001),except for the control group,the expression levels of IRAK1,TRAF6 and NF-κB proteins in the miR agonist group were significantly lower than those in the other three groups,and the expression levels of IκBαprotein were significantly higher than those in the other three groups.Conclusion:Overexpression of miR-146a can negatively regulate corneal inflammatory response in dry eye mice through the IRAK1/TRAF6/NF-κB signaling pathway,which can provide new insights for the clinical treatment of dry eye disease.展开更多
基金supported by the National Natural Science Foundation of China(No.82373380,Xinhua Xie).
文摘Objectives:Deubiquitinase OTUB2 plays a critical role in the progression of various tumors.However,its specific role in triple-negative breast cancer(TNBC)remains unclear.This study aims to elucidate the biological function of OTUB2 in TNBC and uncover the underlying mechanisms.Methods:First,we found that the expression of OTUB2 was upregulated in TNBC by bioinformatics analysis,we then validated its expression in TNBC tissues and cells using immunohistochemistry(IHC)and qPCR and plotted the survival curves by Kaplan-Meier method.Gene set enrichment analysis(GSEA)suggested that OTUB2 may be involved in tumor proliferation and metastasis.Further functional assays,including Cell Counting Kit-8(CCK-8),colony formation,Transwell,and wound healing assays,were performed to assess the effects of OTUB2 overexpression and knockdown on TNBC cell proliferation and migration.Additionally,UbiBrowser 2.0 was used to identify OTUB2 substrate proteins and western blotting was conducted to clarify the molecular mechanisms involved.Results:Our results demonstrated that OTUB2 expression was elevated in TNBC and associated with poor prognosis.Overexpression of OTUB2 enhanced the proliferation and migration of TNBC cells,while its knockdown inhibited these processes.Moreover,OTUB2 stabilized tumor necrosis factor receptor-associated factor 6(TRAF6)by deubiquitinating it,leading to activation of the protein kinase B(AKT)pathway.Conclusions:OTUB2 exerts its promoting effects on the progression of TNBC by activating the TRAF6/AKT pathway.
文摘Objective:To investigate the regulatory effect of miR-146a overexpression on corneal inflammatory response in a mouse model of dry eye,and to analyze its relationship with the IRAK1/TRAF6/NF-кB signaling pathway.Methods:A total of 50 SPF-grade BALB/c mice were randomly divided into five groups,with10 mice in each group.Except for the control group,the other four groups were treated with 0.2%benzalkonium chloride(BAC)solution in both eyes to construct a dry eye model.After successful modeling,the control group and model group received NC agomir;the miR antagonist group received miR-146a antagomir;the miR agonist group received miR-146a agomir;and the pathway agonist group received miR-146a agomir+NF-κB activator 2.After four weeks of treatment,the expressions levels of miR-146a,inflammatory factors,and IRAK1/TRAF6/NF-κB signaling pathway proteins were observed and compared among the five groups.Results:After four weeks of treatment,there was a statistically significant difference in the relative expression of miR-146a in the five groups(F=61.058,P<0.001),which was significantly higher in the miR agonist group than in the other four groups.After4 weeks of treatment,there were statistically significant differences in the expression levels of IL-1β,IL-6,IL-8 and TNF-αin the five groups(F=84.757,103.658,55.477,46.762;P<0.001).After four weeks of treatment,there were statistically significant differences in the protein expression levels of IRAK1,TRAF6,NF-κB and IκBαin the five groups(F=62.975,77.173,67.108,29.381;P<0.001),except for the control group,the expression levels of IRAK1,TRAF6 and NF-κB proteins in the miR agonist group were significantly lower than those in the other three groups,and the expression levels of IκBαprotein were significantly higher than those in the other three groups.Conclusion:Overexpression of miR-146a can negatively regulate corneal inflammatory response in dry eye mice through the IRAK1/TRAF6/NF-κB signaling pathway,which can provide new insights for the clinical treatment of dry eye disease.
