Transcription Factor ELF1 Activates MEIS1 Transcription and Then Regulates the GFI1/FBW7 Axis to Promote the Development of Glioma

Abstract

Glioma is the most common malignancy in the central nervous system with no immediate prospect of a cure. Comprehensive understanding on the pathogenesis of the disorder contributes to a better outcome. Herein, we aimed to investigate whether transcription factors erythroblast transformation-specific (ETS) transcription factor (ELF1), myeloid ecotropic viral integration site 1 (MEIS1), and growth factor independence 1 (GFI1)/F-box/WD repeat-containing protein 7 (FBW7) mediate progression of glioma. ELF1, MEIS1, and GFI1 were upregulated in glioma cells and tissues, as ELF1 was correlated with poor prognosis. Bioinformatics analysis identified the binding between ELF1 and MEIS1 as well as between GFI1 and FBW7, confirmed by chromatin immunoprecipitation (ChIP) experiments. Functional experiment indicated that silencing of ELT1 decreased MEIS1 expression and that overexpression of MEIS1 increased GFI1 expression by activating GFI1 enhancer but decreased FBW7 expression. Importantly, silencing of ELF1 decreased the capacities of proliferation, migration, and invasion of glioma cells whereas it increased apoptosis, supported by increased capase-3 and decreased matrix metalloproteinase-9 (MMP-9) and proliferating cell nuclear antigen (PCNA) expression. Moreover, an in vivo experiment confirmed the inhibitory role of silenced ELF1 in tumor growth, with a decreased level of MEIS1 and GFI1. Taken together, our study elucidated a potential mechanism that ELF1 promoted cell progression by increasing GFI1 and METS1 as well as decreasing FBW7 expression in glioma. Glioma is the most common malignancy in the central nervous system with no immediate prospect of a cure. Comprehensive understanding on the pathogenesis of the disorder contributes to a better outcome. Herein, we aimed to investigate whether transcription factors erythroblast transformation-specific (ETS) transcription factor (ELF1), myeloid ecotropic viral integration site 1 (MEIS1), and growth factor independence 1 (GFI1)/F-box/WD repeat-containing protein 7 (FBW7) mediate progression of glioma. ELF1, MEIS1, and GFI1 were upregulated in glioma cells and tissues, as ELF1 was correlated with poor prognosis. Bioinformatics analysis identified the binding between ELF1 and MEIS1 as well as between GFI1 and FBW7, confirmed by chromatin immunoprecipitation (ChIP) experiments. Functional experiment indicated that silencing of ELT1 decreased MEIS1 expression and that overexpression of MEIS1 increased GFI1 expression by activating GFI1 enhancer but decreased FBW7 expression. Importantly, silencing of ELF1 decreased the capacities of proliferation, migration, and invasion of glioma cells whereas it increased apoptosis, supported by increased capase-3 and decreased matrix metalloproteinase-9 (MMP-9) and proliferating cell nuclear antigen (PCNA) expression. Moreover, an in vivo experiment confirmed the inhibitory role of silenced ELF1 in tumor growth, with a decreased level of MEIS1 and GFI1. Taken together, our study elucidated a potential mechanism that ELF1 promoted cell progression by increasing GFI1 and METS1 as well as decreasing FBW7 expression in glioma. IntroductionMalignant gliomas are among the most common primary brain tumors in adults.1Ostrom Q.T. Gittleman H. Truitt G. Boscia A. Kruchko C. Barnholtz-Sloan J.S. CBTRUS statistical report: primary brain and other central nervous system tumors diagnosed in the united states in 2011–2015.Neuro-oncol. 2018; 20: iv1-iv86Crossref PubMed Scopus (1156) Google Scholar Due to the infiltrative nature of this disease and the localization close to eloquent brain areas, surgical resection fails to cure the disease. Patients diagnosed with a malignant glioma such as glioblastoma have to undergo a fierce clinical course with a survival time of less than 2 years for most patients.2Oike T. Suzuki Y. Sugawara K. Shirai K. Noda S.E. Tamaki T. Nagaishi M. Yokoo H. Nakazato Y. Nakano T. Radiotherapy plus concomitant adjuvant temozolomide for glioblastoma: Japanese mono-institutional results.PLoS ONE. 2013; 8: e78943Crossref PubMed Scopus (65) Google Scholar,3Lesueur P. Lequesne J. Grellard J.M. Dugué A. Coquan E. Brachet P.E. Geffrelot J. Kao W. Emery E. Berro D.H. et al.Phase I/IIa study of concomitant radiotherapy with olaparib and temozolomide in unresectable or partially resectable glioblastoma: OLA-TMZ-RTE-01 trial protocol.BMC Cancer. 2019; 19: 198Crossref PubMed Scopus (58) Google Scholar Based on advances in the molecular characterization of these tumors, disease-associated targets, including epidermal growth factor receptor (EGFR) or vascular EGFR (VEGFR), were identified, which led to the development of new approaches using traditional routes of drug development.4Chinot O.L. Wick W. Mason W. Henriksson R. Saran F. Nishikawa R. Carpentier A.F. Hoang-Xuan K. Kavan P. Cernea D. et al.Bevacizumab plus radiotherapy-temozolomide for newly diagnosed glioblastoma.N. Engl. J. Med. 2014; 370: 709-722Crossref PubMed Scopus (1645) Google Scholar, 5Gilbert M.R. Dignam J.J. Armstrong T.S. Wefel J.S. Blumenthal D.T. Vogelbaum M.A. Colman H. Chakravarti A. Pugh S. Won M. et al.A randomized trial of bevacizumab for newly diagnosed glioblastoma.N. Engl. J. Med. 2014; 370: 699-708Crossref PubMed Scopus (1779) Google Scholar, 6Boxerman J.L. Zhang Z. Safriel Y. Rogg J.M. Wolf R.L. Mohan S. Marques H. Sorensen A.G. Gilbert M.R. Barboriak D.P. Prognostic value of contrast enhancement and FLAIR for survival in newly diagnosed glioblastoma treated with and without bevacizumab: results from ACRIN 6686.Neuro-oncol. 2018; 20: 1400-1410Crossref PubMed Scopus (18) Google Scholar According to the World Health Organization (WHO) classification of tumors of the central nervous system, gliomas can be categorized into four grades (grades I–IV), among which grade IV is also called glioblastoma or glioblastoma multiforme (GBM).7Rogers T.W. Toor G. Drummond K. Love C. Field K. Asher R. Tsui A. Buckland M. Gonzales M. The 2016 revision of the WHO Classification of Central Nervous System Tumours: retrospective application to a cohort of diffuse gliomas.J. Neurooncol. 2018; 137: 181-189Crossref PubMed Scopus (30) Google Scholar Moreover, a gene expression-based molecular classification of glioblastoma has been presented, including proneural, neural, classical, and mesenchymal subtypes.8Brat T. of the WHO Classification of of the Central Nervous and PubMed Scopus Google Scholar the of these no for gliomas has been in to these the of the disease was and factors such as the of to the or to the tumor were is by the that of in gene expression a tumor a of D.H. P. P. S. G. et and of and and and 2018; PubMed Scopus Google transformation-specific (ETS) transcription factors in with as and as tumor factors for binding in expression of can have that transcription factor 1 as a factor for tumor E. A. W. C. P. A. S. A. analysis of 2013; PubMed Scopus Google Scholar in cell indicated that ELF1 has tumor by ELF1 and transcription factors to also a in cell M. M. T. D. K. J. K. T. H. K. et and of transcription factors in PubMed Scopus Google Scholar of ELF1 increased that the in tumors of in glioma and can to the transcription factor myeloid ecotropic viral integration site 1 (MEIS1), and the of MEIS1 to growth factor independence 1 in of proliferation, migration, and invasion and of cell in glioma ELF1 in and Cancer. 2018; PubMed Scopus Google Scholar MEIS1, a transcription in cell development and cell C. H. M. J.M. G. J. et and expression and a role for binding in 2018; PubMed Scopus Google Scholar GFI1 is in the and as a GFI1 as a by or with other K. F. in 2019; PubMed Scopus Google Scholar The tumor of these factors and with factors are well our we aimed to investigate the mechanism ELF1 the progression of glioma. results that of ELF1 in glioma to the transcription factor MEIS1 and the of MEIS1 to GFI1 enhancer in glioma an was also to the of on tumor in Glioma and with WHO and of and were by and we and that were in these four using the transcription factors were from and including ELF1, and among which ELF1 has been as with M. C. J. Y. S. D. Z. H. Y. of glioma Med. PubMed Scopus Google Scholar The expression from and were in a ELF1 was indicated as in analysis of from also identified the expression of ELF1 to whether ELF1 was in the and development of the expression of ELF1 in brain of glioma and brain was by with the the expression of ELF1 in glioma was increased The expression of ELF1 increased with the of the WHO grade of glioma we the ELF1 expression and the between the clinical According to the ELF1 expression in gliomas it was into a and The results that the expression of ELF1 an with the WHO classification and in was no between expression of ELF1 and tumor and tumor the of survival that expression of ELF1 was correlated with survival time and of between ELF1 and the of Glioma in a new ELF1 the and of Glioma and of the of ELF1 in glioma tissues, in to it the proliferation, migration, and invasion of glioma we on glioma to ELF1 and the most on ELF1 to the results from with the expression of ELF1 was decreased in glioma cells and to results of and analysis the results of a and glioma cell of proliferation, migration, and invasion were and cell was of ELF1 factor proliferating cell nuclear antigen factor matrix metalloproteinase-9 and factor expression was by and results that with the of and in the were of were increased ELF1 and and of and Glioma of ELF1 in and glioma cells or was by and of of and cells with or of of and cells with or of of and cells with or of and cells was by analysis was to the expression of factor factor and factor of and cells with or The are as with the was between the and the of time were The cell experiment was ELF1 to the MEIS1 to and the of the we have identified that ELF1 was in glioma and can the proliferation, migration, and invasion of glioma cells we our of and that ELF1, a transcription can be into MEIS1 the P. C. A. S. D. of factor 1 as a of the PubMed Scopus Google Scholar The of ELF1 and MEIS1 of from and identified a between ELF1 and MEIS1 analysis confirmed the analysis of from The The analysis of the four also that MEIS1 was in glioma in the and it was in and but Based on this we that ELF1 glioma development by the transcription of MEIS1 expression in glioma the and analysis and identified the expression of MEIS1 in glioma to brain The binding site between ELF1 and MEIS1 by the was by a chromatin immunoprecipitation (ChIP) protein to and that with the the of binding MEIS1 in the ELF1 was increased were into glioma and protein of MEIS1 were decreased to the MEIS1 to the of MEIS1 to in the of expression of ELF1 and MEIS1 from and of ELF1 and MEIS1 analysis of from of MEIS1 expression from and as well as from and analysis of MEIS1 expression in clinical glioma and brain site of ELF1 and METS1 the experiment of METS1 level in the and analysis of MEIS1 protein expression in and of of and cells with and of of and cells with and of invasion of and cells with and of and cells with and analysis of and capase-3 protein expression in and cells with and with or with or The between were by an The among were by with time among were by by a The cell experiment was to whether ELF1 was in the development of glioma the transcription of MEIS1, we overexpression and into glioma cells and by and as well as was that with decreased cell proliferation, migration, and invasion whereas it increased apoptosis, in with that of the Based on of the of ELF1 on glioma the growth, migration, and invasion of glioma and cell analysis was to factor protein and factor The expression of and in the was decreased and the expression of was overexpression of MEIS1 increased the expression of and and decreased the expression of these factors results that the transcription factor ELF1 be in glioma development by MEIS1 transcription in Glioma by the of the GFI1 a gene expression and by in the study to a between MEIS1 and GFI1 this in tissues, we GFI1 expression in the glioma and by and and that expression of GFI1 in glioma was glioma cells were with overexpression of MEIS1, as by GFI1 expression in cells was increased the experiment was to the between MEIS1 and GFI1. of MEIS1 in and cells led to the of and MEIS1 in the GFI1 and enhancer with results in the cell results indicated that MEIS1 promoted the expression of GFI1 by activating the enhancer of GFI1 and and GFI1 in and Glioma analysis of between MEIS1 and GFI1. analysis of GFI1 expression in glioma and brain analysis of GFI1 expression in and cells with or of of and MEIS1 in the GFI1 enhancer with or the of and MEIS1 in the of GFI1 with or of and cell with or of of and cells with or of invasion of and cells with or of of and cells with or analysis of and capase-3 protein expression in and cells with or of GFI1 and expression of and of expression from and as well as from and analysis of between GFI1 and analysis of FBW7 protein expression in glioma cells are as The between were by an The among were by with time among were by by a The cell experiment was the between MEIS1 and GFI1 on we treated cells with MEIS1 or GFI1 by a and with the cell proliferation, migration, and invasion were increased but the was decreased in the The of GFI1 were and with of was to to cells the of MEIS1 on cell progression and results were supported by the of and in the cells in and with GFI1 the of overexpression of MEIS1 on the expression of and and the expression of the of as GFI1 can the expression of repeat-containing protein 7 (FBW7) with H. Zhang F. Z. of of FBW7 2018; PubMed Scopus Google Scholar the the expression for FBW7 in we the FBW7 from the other to the between GFI1 and that was a between GFI1 and FBW7 from the and indicated expression of FBW7 in with the results from and by GFI1 and FBW7 a as by analysis we to the expression of FBW7 in glioma cells by analysis with MEIS1 or GFI1. in and with the FBW7 expression was in the but with expression of elucidated a mechanism that MEIS1 FBW7 expression the of GFI1 and proliferation, migration, and invasion and of glioma with ELF1 Glioma by the the in vivo of ELF1, we a we silenced ELF1 expression in cells and the cells into the we the and of the The results that the and of tumor in treated with were than in the analysis that the expression of ELF1, MEIS1, and with expression of FBW7 results indicated that with ELF1 expression of FBW7 and tumor growth in in of tumor with or of tumor growth in with or of with or analysis of ELF1, MEIS1, and FBW7 with or with the are as The between were by an time among were by by a The cell experiment was the most common of primary malignant brain and for and cell are by a and survival of years and the survival is with of and T. of the WHO Classification of of the Central Nervous and PubMed Scopus Google Scholar the we that of ELF1 in glioma can to the transcription factor MEIS1 and the of MEIS1 to GFI1 in the of proliferation, migration, and invasion and of cell in glioma was identified as a potential of the and cells with a ELF1 were to cell by the J.J. Y. J.M. of the protein in PubMed Scopus Google Scholar, binding between tumor transcription factors and 2018; PubMed Scopus Google Scholar, S. A. J. Z. Y. et and analysis protein to PubMed Scopus Google Scholar the of whether ELF1 was in the and development of we the expression of ELF1 in brain of glioma and that ELF1 is in glioma and with WHO and the of we that ELF1 as a factor of tumor to our we silenced the ELF1 in and cell and that the of cells was silencing of ELF1 in and cell of and invasion of glioma cells were by silencing of The factor and factor were factor was upregulated of ELF1 in glioma The results that silencing ELF1 the proliferation, migration, and invasion and cell of glioma is a transcription factor that in cell development and cell C. H. M. J.M. G. J. et and expression and a role for binding in 2018; PubMed Scopus Google Scholar MEIS1 has a role in the of the of cells and the transcription of as well as in cell development and an role in A. M.R. MEIS1 of cell Med. 2019; PubMed Scopus Google Scholar has been that ELF1 can as an of the P. C. A. S. D. of factor 1 as a of the PubMed Scopus Google Scholar we that ELF1 glioma development by the transcription of Based on the results from the expression of MEIS1 in glioma was upregulated in to overexpression of MEIS1 the of ELF1 on the growth, migration, and invasion of glioma and it cell in glioma can be that the transcription factor ELF1 be in glioma progression by MEIS1 GFI1 which is a transcription factor for development of the and was in the system, it as a of cell as well as development of the and G. A. A. S. of by and and ONE. 2013; 8: PubMed Scopus Google J. K. expression is by with and as in PubMed Scopus Google Scholar study that GFI1 expression is by with and MEIS1 as in J. K. expression is by with and as in PubMed Scopus Google Scholar the mechanism between GFI1 and MEIS1 in glioma analysis was to that a of between MEIS1 and GFI1 of MEIS1 in glioma cells the of and MEIS1 in the GFI1 enhancer as well as the of and that MEIS1 promoted the expression of GFI1 by activating the enhancer of GFI1. of MEIS1 the enhancer of and and and in glioma Moreover, that the tumor FBW7, an that and of J. M. G. Y. and cell progression by Cancer. 2019; PubMed Scopus Google Scholar in and promoted in the of in glioma MEIS1 the of the GFI1 by of the expression of FBW7 and of the proliferation, migration, and invasion and of of glioma a tumor of cells of was to the of ELF1 in The results that the growth and of tumors in treated with were than in the with of ELF1, MEIS1, and and upregulated expression of FBW7, were in tumor treated with The results that with ELF1 expression of FBW7 and glioma growth in the study that upregulated expression of ELF1 in glioma promoted tumor progression by the that ELF1 as a for the of ELF1 in expression of ELF1 in glioma tumor progression by the that ELF1 as a for and were using the and from the The D. Y. W. expression for and PubMed Scopus Google Scholar for was for expression analysis with the as and value The of the was to a of the and was to the of the with a and the The expression a of including and 7 glioma 7 and glioma 2 and glioma and and glioma in these expression were by analysis R. S. P. J. for gene integration and PubMed Scopus Google Scholar and were to transcription The the of and transcription factors were as transcription The were the and the gene of transcription factor was from the of The binding site of transcription factor and gene was the gene expression of ELF1, MEIS1, and GFI1 from the of the and by with and a to the The of the was by analysis and ELF1, MEIS1, and GFI1 expression in glioma was using of and brain of the with glioma confirmed by were from from to According to the 2016 WHO classification of central nervous system A. G. A. D. H. P. The 2016 World Health Organization Classification of of the Central Nervous a PubMed Scopus Google Scholar gliomas were into four grades and the in this study of of grade and of grade with of and of were as with other malignant tumor clinical and of brain by to were as the with to by or the of were to and the was The survival of was in this study an and were by our to with the of and cell and by of were in and in were to the the cell and was in the growth cells were with into cell and on for were using gene overexpression and the expression gene silencing and were by was in which were in and other the and cells were in the growth were by and and cell were on and the was to the cells for and cells with were and for experiments. The of was and the of was was to the of and the was into using with a system of the was to on the for the The were to using a system as an the was to the gene expression. were to for in a new and 1 cells were treated with 1 of cell for and for and to the the protein of was using a the were to a the was in for 2 and with with time for The was with the primary ELF1 MEIS1 GFI1 and The the was with time for was and for 1 The was with for in for 1 and to in the and the protein were was as and the of value of and was as the expression of and cells were and The cell was to 1 and the cells were into a with and were treated to the of the and cell was by and of was in the for and was with an the of the growth the invasion the was into a of was to of which of was to and for the were and and the cell was with of cell was to the of and of cell was to the were for a was in for and with The cells were with for and the cells on the were with on the were and by an was for the and the time was from four were for the cells were with and in the and the was and were into a of to the of 1 cells were in of and the cells were and for 1 of was to the for and and were by of and the of to cell was for According to the the cells were with and with for to The cells were with a cell and a nuclear and treated with to of chromatin of the as were by protein with or was to the protein was with the was by were from or was into the cell was in the treated with with were and with a The of for tumor is a a is of tumor and is of were to and tumors of were and of the were by the and and of the in this study are in with the and for tumor were to and and were in for with for and with antigen for and were with for and with primary were with for were to with for and by an and were from and cells were in The of cells was and the of cells was The results were by in a of the were using and as of experiments. The between were by an The among were by analysis of with time among were by or survival was by the and the between was by was for was IntroductionMalignant gliomas are among the most common primary brain tumors in adults.1Ostrom Q.T. Gittleman H. Truitt G. Boscia A. Kruchko C. Barnholtz-Sloan J.S. CBTRUS statistical report: primary brain and other central nervous system tumors diagnosed in the united states in 2011–2015.Neuro-oncol. 2018; 20: iv1-iv86Crossref PubMed Scopus (1156) Google Scholar Due to the infiltrative nature of this disease and the localization close to eloquent brain areas, surgical resection fails to cure the disease. Patients diagnosed with a malignant glioma such as glioblastoma have to undergo a fierce clinical course with a survival time of less than 2 years for most patients.2Oike T. Suzuki Y. Sugawara K. Shirai K. Noda S.E. Tamaki T. Nagaishi M. Yokoo H. Nakazato Y. Nakano T. Radiotherapy plus concomitant adjuvant temozolomide for glioblastoma: Japanese mono-institutional results.PLoS ONE. 2013; 8: e78943Crossref PubMed Scopus (65) Google Scholar,3Lesueur P. Lequesne J. Grellard J.M. Dugué A. Coquan E. Brachet P.E. Geffrelot J. Kao W. Emery E. Berro D.H. et al.Phase I/IIa study of concomitant radiotherapy with olaparib and temozolomide in unresectable or partially resectable glioblastoma: OLA-TMZ-RTE-01 trial protocol.BMC Cancer. 2019; 19: 198Crossref PubMed Scopus (58) Google Scholar Based on advances in the molecular characterization of these tumors, disease-associated targets, including epidermal growth factor receptor (EGFR) or vascular EGFR (VEGFR), were identified, which led to the development of new approaches using traditional routes of drug development.4Chinot O.L. Wick W. Mason W. Henriksson R. Saran F. Nishikawa R. Carpentier A.F. Hoang-Xuan K. Kavan P. Cernea D. et al.Bevacizumab plus radiotherapy-temozolomide for newly diagnosed glioblastoma.N. Engl. J. Med. 2014; 370: 709-722Crossref PubMed Scopus (1645) Google Scholar, 5Gilbert M.R. Dignam J.J. Armstrong T.S. Wefel J.S. Blumenthal D.T. Vogelbaum M.A. Colman H. Chakravarti A. Pugh S. Won M. et al.A randomized trial of bevacizumab for newly diagnosed glioblastoma.N. Engl. J. Med. 2014; 370: 699-708Crossref PubMed Scopus (1779) Google Scholar, 6Boxerman J.L. Zhang Z. Safriel Y. Rogg J.M. Wolf R.L. Mohan S. Marques H. Sorensen A.G. Gilbert M.R. Barboriak D.P. Prognostic value of contrast enhancement and FLAIR for survival in newly diagnosed glioblastoma treated with and without bevacizumab: results from ACRIN 6686.Neuro-oncol. 2018; 20: 1400-1410Crossref PubMed Scopus (18) Google Scholar According to the World Health Organization (WHO) classification of tumors of the central nervous system, gliomas can be categorized into four grades (grades I–IV), among which grade IV is also called glioblastoma or glioblastoma multiforme (GBM).7Rogers T.W. Toor G. Drummond K. Love C. Field K. Asher R. Tsui A. Buckland M. Gonzales M. The 2016 revision of the WHO Classification of Central Nervous System Tumours: retrospective application to a cohort of diffuse gliomas.J. Neurooncol. 2018; 137: 181-189Crossref PubMed Scopus (30) Google Scholar Moreover, a gene expression-based molecular classification of glioblastoma has been presented, including proneural, neural, classical, and mesenchymal subtypes.8Brat T. of the WHO Classification of of the Central Nervous and PubMed Scopus Google Scholar the of these no for gliomas has been in to these the of the disease was and factors such as the of to the or to the tumor were is by the that of in gene expression a tumor a of D.H. P. P. S. G. et and of and and and 2018; PubMed Scopus Google transformation-specific (ETS) transcription factors in with as and as tumor factors for binding in expression of can have that transcription factor 1 as a factor for tumor E. A. W. C. P. A. S. A. analysis of 2013; PubMed Scopus Google Scholar in cell indicated that ELF1 has tumor by ELF1 and transcription factors to also a in cell M. M. T. D. K. J. K. T. H. K. et and of transcription factors in PubMed Scopus Google Scholar of ELF1 increased that the in tumors of in glioma and can to the transcription factor myeloid ecotropic viral integration site 1 (MEIS1), and the of MEIS1 to growth factor independence 1 in of proliferation, migration, and invasion and of cell in glioma ELF1 in and Cancer. 2018; PubMed Scopus Google Scholar MEIS1, a transcription in cell development and cell C. H. M. J.M. G. J. et and expression and a role for binding in 2018; PubMed Scopus Google Scholar GFI1 is in the and as a GFI1 as a by or with other K. F. in 2019; PubMed Scopus Google Scholar The tumor of these factors and with factors are well our we aimed to investigate the mechanism ELF1 the progression of glioma. results that of ELF1 in glioma to the transcription factor MEIS1 and the of MEIS1 to GFI1 enhancer in glioma an was also to the of on tumor

Topics & Concepts