FK506-binding protein 9(FKBP9)is involved in tumor malignancy by resistance to endoplasmic reticulum(ER)stress,and the up-regulation of FKBP9 is associated with patients'poor prognosis.The current knowledge of the...FK506-binding protein 9(FKBP9)is involved in tumor malignancy by resistance to endoplasmic reticulum(ER)stress,and the up-regulation of FKBP9 is associated with patients'poor prognosis.The current knowledge of the molecular mechanisms is still limited.One pre-vious study showed that FKBP9 could confer glioblastoma cell resistance to ER stress through ASK1-p38 signaling.However,the upstream regulatory mechanism of FKBP9 expression is still indistinct.In this study,we identified the FKBP9 binding proteins using co-immunoprecipitation followed by mass spectrometry.Results showed that FKBP9 interacted with the binding immu-noglobulin protein(BiP).BiP bound directly to FKBP9 with high affinity.BiP prolonged the half-life of the FKBP9 protein and stabilized the FKBP9 protein.BiP and FKBP9 protein levels were positively correlated in patients with glioma,and patients with high expression of BiP and FKBP9 showed a worse prognosis.Further studies showed that FKBP9 knockout in genetically engineered mice inhibited intracranial glioblastoma formation and prolonged survival by decreasing cellular proliferation and ER stress-induced CHOP-related apoptosis.Moreover,normal cells may depend less on FKBP9,as shown by the absence of apoptosis upon FKBP9 knockdown in a non-transformed human cell line and overall normal development in homozygous knockout mice.These findings suggest an important role of BiP-regulated FKBP9-associated signaling in glioma progression and the BiP-FKBP9 axis may be a potential therapeutic target forglioma.展开更多
Background:Long non-coding RNAs(lncRNAs)play key roles in human cancers.In our previous study,we demonstrated that lncRNA FKBP prolyl isomerase 9 pseudogene 1(FKBP9P1)was highly expressed in head and neck squamous cel...Background:Long non-coding RNAs(lncRNAs)play key roles in human cancers.In our previous study,we demonstrated that lncRNA FKBP prolyl isomerase 9 pseudogene 1(FKBP9P1)was highly expressed in head and neck squamous cell cancer(HNSCC)tissues.However,its functional significance remains poorly understood.In the present study,we identify the role and potential molecular biologic mechanisms of FKBP9P1 in HNSCC.Methods:Quantitative real-time polymerase chain reaction was used to detect the expression of FKBP9P1 in HNSCC tissues,matched adjacent normal tissues,human HNSCC cells(FaDu,Cal-27,SCC4,and SCC9),and human immortalized keratinocytes cell HaCaT(normal control).Cal-27 and SCC9 cells were transfected with sh-FKBP9P1-1,sh-FKBP9P1-2,and normal control(sh-NC)lentivirus.Cell counting kit-8 assay,colony formation assay,wound healing assay,and trans-well assay were used to explore the biologic function of FKBP9P1 in HNSCC cells.Furthermore,western blotting was used to determine the mechanism of FKBP9P1 in HNSCC progression.Chi-squared test was performed to assess the clinical significance among FKBP9P1 high-expression and low-expression groups.Survival analyses were performed using the Kaplan-Meier method and assessed using the log-rank test.The comparison between two groups was analyzed by Student t test,and comparisons among multiple samples were performed by one-way analysis of variance and a Bonferroni post hoc test.Results:FKBP9P1 expression was significantly up-regulated in HNSCC tissues(tumor vs.normal,1.914 vs.0.957,t=7.746,P<0.001)and cell lines(P<0.01 in all HNSCC cell lines).Besides,the median FKBP9P1 expression of HNSCC tissues(1.677)was considered as the threshold.High FKBP9P1 level was correlated with advanced T stage(P=0.022),advanced N stage(P=0.036),advanced clinical stage(P=0.018),and poor prognosis of HNSCC patients(overall survival,P=0.002 and disease-free survival,P<0.001).Knockdown of FKBP9P1 led to marked repression in proliferation,migration,and invasion of HNSCC cells in vitro(P all<0.01).Mechanistically,silencing FKBP9P1 was observed to restrain the PI3K/AKT signaling pathway.Conclusions:Silencing lncRNA FKBP9P1 represses HNSCC progression and inhibits PI3K/AKT(phosphatidylinositol 3 kinase/AKT Serine/Threonine Kinase)signaling in vitro.Therefore,FKBP9P1 could be a potential new target for the diagnosis and treatment of HNSCC patients.展开更多
基金supported by the National Natural Science Foundation of China (No.82103107 to B.S.)the open project from the State Key Laboratory of Genetic Resources and Evolution of China (No.GREKF19-06 to H.Y.)the 1.3.5 project for disciplines of excellence,West China Hospital,Sichuan University (No.ZYYC20002 to X.D.Z.).
文摘FK506-binding protein 9(FKBP9)is involved in tumor malignancy by resistance to endoplasmic reticulum(ER)stress,and the up-regulation of FKBP9 is associated with patients'poor prognosis.The current knowledge of the molecular mechanisms is still limited.One pre-vious study showed that FKBP9 could confer glioblastoma cell resistance to ER stress through ASK1-p38 signaling.However,the upstream regulatory mechanism of FKBP9 expression is still indistinct.In this study,we identified the FKBP9 binding proteins using co-immunoprecipitation followed by mass spectrometry.Results showed that FKBP9 interacted with the binding immu-noglobulin protein(BiP).BiP bound directly to FKBP9 with high affinity.BiP prolonged the half-life of the FKBP9 protein and stabilized the FKBP9 protein.BiP and FKBP9 protein levels were positively correlated in patients with glioma,and patients with high expression of BiP and FKBP9 showed a worse prognosis.Further studies showed that FKBP9 knockout in genetically engineered mice inhibited intracranial glioblastoma formation and prolonged survival by decreasing cellular proliferation and ER stress-induced CHOP-related apoptosis.Moreover,normal cells may depend less on FKBP9,as shown by the absence of apoptosis upon FKBP9 knockdown in a non-transformed human cell line and overall normal development in homozygous knockout mice.These findings suggest an important role of BiP-regulated FKBP9-associated signaling in glioma progression and the BiP-FKBP9 axis may be a potential therapeutic target forglioma.
基金This work was supported by grants from Beijing Municipal Natural Science Foundation(No.7184196)Beijing Municipal Administration of Hospitals’Ascent Plan(No.DFL20180202)+1 种基金Capital Health Development Research Project(No.Shoufa-2018-2-2054)Beijing Natural Science Foundation Program and Scientific Research Key Program of Beijing Municipal Commission of Education(No.KZ201910025034)。
文摘Background:Long non-coding RNAs(lncRNAs)play key roles in human cancers.In our previous study,we demonstrated that lncRNA FKBP prolyl isomerase 9 pseudogene 1(FKBP9P1)was highly expressed in head and neck squamous cell cancer(HNSCC)tissues.However,its functional significance remains poorly understood.In the present study,we identify the role and potential molecular biologic mechanisms of FKBP9P1 in HNSCC.Methods:Quantitative real-time polymerase chain reaction was used to detect the expression of FKBP9P1 in HNSCC tissues,matched adjacent normal tissues,human HNSCC cells(FaDu,Cal-27,SCC4,and SCC9),and human immortalized keratinocytes cell HaCaT(normal control).Cal-27 and SCC9 cells were transfected with sh-FKBP9P1-1,sh-FKBP9P1-2,and normal control(sh-NC)lentivirus.Cell counting kit-8 assay,colony formation assay,wound healing assay,and trans-well assay were used to explore the biologic function of FKBP9P1 in HNSCC cells.Furthermore,western blotting was used to determine the mechanism of FKBP9P1 in HNSCC progression.Chi-squared test was performed to assess the clinical significance among FKBP9P1 high-expression and low-expression groups.Survival analyses were performed using the Kaplan-Meier method and assessed using the log-rank test.The comparison between two groups was analyzed by Student t test,and comparisons among multiple samples were performed by one-way analysis of variance and a Bonferroni post hoc test.Results:FKBP9P1 expression was significantly up-regulated in HNSCC tissues(tumor vs.normal,1.914 vs.0.957,t=7.746,P<0.001)and cell lines(P<0.01 in all HNSCC cell lines).Besides,the median FKBP9P1 expression of HNSCC tissues(1.677)was considered as the threshold.High FKBP9P1 level was correlated with advanced T stage(P=0.022),advanced N stage(P=0.036),advanced clinical stage(P=0.018),and poor prognosis of HNSCC patients(overall survival,P=0.002 and disease-free survival,P<0.001).Knockdown of FKBP9P1 led to marked repression in proliferation,migration,and invasion of HNSCC cells in vitro(P all<0.01).Mechanistically,silencing FKBP9P1 was observed to restrain the PI3K/AKT signaling pathway.Conclusions:Silencing lncRNA FKBP9P1 represses HNSCC progression and inhibits PI3K/AKT(phosphatidylinositol 3 kinase/AKT Serine/Threonine Kinase)signaling in vitro.Therefore,FKBP9P1 could be a potential new target for the diagnosis and treatment of HNSCC patients.
