Objective:BYSL,which encodes the Bystin protein in humans,is upregulated in reactive astrocytes following brain damage and/or inflammation.We aimed to determine the role and mechanism of BYSL in glioma cell growth and...Objective:BYSL,which encodes the Bystin protein in humans,is upregulated in reactive astrocytes following brain damage and/or inflammation.We aimed to determine the role and mechanism of BYSL in glioma cell growth and survival.Methods:BYSL expression in glioma tissues was measured by quantitative real-time PCR,Western blot,and immunohistochemistry.In vitro assays were performed to assess the role of BYSL in cell proliferation and apoptosis.Protein interactions and co-localization were determined by co-immunoprecipitation and double immunofluorescence.The expression and activity of the AKT/m TOR signaling molecules were determined by Western blot analysis,and the role of BYSL in glioma growth was confirmed in an orthotopic xenograft model.Results:The BYSL m RNA and protein levels were elevated in glioma tissues.Silencing BYSL inhibited glioma cell proliferation,impeded cell cycle progression,and induced apoptosis,whereas overexpressing BYSL protein led to the opposite effects.We identified a complex consisting of BYSL,RIOK2,and m TOR,and observed co-localization and positive correlations between BYSL and RIOK2 in glioma cells and tissues.Overexpressing BYSL or RIOK2 increased the expression and activity of AKT/m TOR signaling molecules,whereas downregulation of BYSL or RIOK2 decreased the activity of AKT/m TOR signaling molecules.Silencing BYSL or RIOK2 decreased the growth of the tumors and prolonged the lifespan of the animals in an orthotopic xenograft model.Conclusions:High expression of BYSL in gliomas promoted tumor cell growth and survival both in vitro and in vivo.These effects could be attributed to the association of BYSL with RIOK2 and m TOR,and the subsequent activation of AKT signaling.展开更多
基金supported by the National Natural Science Foundation of China(Grant Nos.82002632 and 82072763)the Key Research&Development Plan of Jiangsu Province(Grant No.BE2017636)+3 种基金Xuzhou City(Grant No.KC20076)supported by the Six Talents Peak(Grant No.2019-SWYY-092)the Medical Youth Talent(Grant No.QNRC2016787)the Qing Lan projects of Jiangsu Province。
文摘Objective:BYSL,which encodes the Bystin protein in humans,is upregulated in reactive astrocytes following brain damage and/or inflammation.We aimed to determine the role and mechanism of BYSL in glioma cell growth and survival.Methods:BYSL expression in glioma tissues was measured by quantitative real-time PCR,Western blot,and immunohistochemistry.In vitro assays were performed to assess the role of BYSL in cell proliferation and apoptosis.Protein interactions and co-localization were determined by co-immunoprecipitation and double immunofluorescence.The expression and activity of the AKT/m TOR signaling molecules were determined by Western blot analysis,and the role of BYSL in glioma growth was confirmed in an orthotopic xenograft model.Results:The BYSL m RNA and protein levels were elevated in glioma tissues.Silencing BYSL inhibited glioma cell proliferation,impeded cell cycle progression,and induced apoptosis,whereas overexpressing BYSL protein led to the opposite effects.We identified a complex consisting of BYSL,RIOK2,and m TOR,and observed co-localization and positive correlations between BYSL and RIOK2 in glioma cells and tissues.Overexpressing BYSL or RIOK2 increased the expression and activity of AKT/m TOR signaling molecules,whereas downregulation of BYSL or RIOK2 decreased the activity of AKT/m TOR signaling molecules.Silencing BYSL or RIOK2 decreased the growth of the tumors and prolonged the lifespan of the animals in an orthotopic xenograft model.Conclusions:High expression of BYSL in gliomas promoted tumor cell growth and survival both in vitro and in vivo.These effects could be attributed to the association of BYSL with RIOK2 and m TOR,and the subsequent activation of AKT signaling.
