title: Inhibition of REST Suppresses Proliferation and Migration in Glioblastoma Cells.
abstract: Glioblastoma (GBM) is the most common primary brain tumor, with poor prognosis and a lack of effective therapeutic options. The aberrant expression of transcription factor REST (repressor element 1-silencing transcription factor) had been reported in different kinds of tumors. However, the function of REST and its mechanisms in GBM remain elusive. Here, REST expression was inhibited by siRNA silencing in U-87 and U-251 GBM cells. Then CCK-8 assay showed significantly decreased cell proliferation, and the inhibition of migration was verified by scratch wound healing assay and transwell assay. Using cell cycle analysis and Annexin V/PI straining assay, G1 phase cell cycle arrest was found to be a reason for the suppression of cell proliferation and migration upon REST silencing, while apoptosis was not affected by REST silencing. Further, the detection of REST-downstream genes involved in cytostasis and migration inhibition demonstrated that CCND1 and CCNE1 were reduced; CDK5R1, BBC3, EGR1, SLC25A4, PDCD7, MAPK11, MAPK12, FADD and DAXX were enhanced, among which BBC3 and DAXX were direct targets of REST, as verified by ChIP (chromatin immunoprecipitation) and Western blotting. These data suggested that REST is a master regulator that maintains GBM cells proliferation and migration, partly through regulating cell cycle by repressing downstream genes, which might represent a potential target for GBM therapy.
authors.affiliation: Department of Stem Cells and Regenerative Medicine, Shenyang Key Laboratory for Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology, Ministry of Public Health, and Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical University, Shenyang 110122, China. zhangdianbao@gmail.com.
authors.forename: Dianbao
authors.initials: D
authors.lastname: Zhang
authors.affiliation: Department of Stem Cells and Regenerative Medicine, Shenyang Key Laboratory for Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology, Ministry of Public Health, and Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical University, Shenyang 110122, China. liy9012@126.com.
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Stem Cells and Regenerative Medicine, Shenyang Key Laboratory for Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology, Ministry of Public Health, and Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical University, Shenyang 110122, China. wangruicmu@163.com.
authors.forename: Rui
authors.initials: R
authors.lastname: Wang
authors.affiliation: Pharmaceutical Preparation Section, Shenyang Children's Hospital, Shenyang 110032, China. yunnali@sina.cn.
authors.forename: Yunna
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of General Practice, the First Affiliated Hospital of China Medical University, Shenyang 110015, China. 13998309428@163.com.
authors.forename: Ping
authors.initials: P
authors.lastname: Shi
authors.affiliation: Department of Pharmacy, the First Affiliated Hospital of China Medical University, Shenyang 110015, China. zhoumi1018@sina.com.
authors.forename: Zhoumi
authors.initials: Z
authors.lastname: Kan
authors.affiliation: Department of Stem Cells and Regenerative Medicine, Shenyang Key Laboratory for Stem Cells and Regenerative Medicine, Key Laboratory of Cell Biology, Ministry of Public Health, and Key Laboratory of Medical Cell Biology, Ministry of Education, China Medical University, Shenyang 110122, China. pangxining@126.com.|Science Experiment Center, China Medical University, Shenyang 110122, China. pangxining@126.com.
authors.forename: Xining
authors.initials: X
authors.lastname: Pang
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D056970:Co-Repressor Proteins; D019938:Cyclin D1 / Q000235:genetics / Q000378:metabolism; D019927:Cyclin E / Q000235:genetics / Q000378:metabolism; D051766:Early Growth Response Protein 1 / Q000235:genetics / Q000378:metabolism; D053200:Fas-Associated Death Domain Protein / Q000235:genetics / Q000378:metabolism; D016193:G1 Phase; D005909:Glioblastoma / Q000378:metabolism*; D006801:Humans; D026901:Membrane Transport Proteins; D020928:Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D015513:Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D000099229:RE1-Silencing Transcription Factor
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; D002352:Carrier Proteins; D056970:Co-Repressor Proteins; D019938:Cyclin D1; D019927:Cyclin E; D051766:Early Growth Response Protein 1; D053200:Fas-Associated Death Domain Protein; D026901:Membrane Transport Proteins; D020928:Mitogen-Activated Protein Kinases; D018832:Molecular Chaperones; D009419:Nerve Tissue Proteins; D009687:Nuclear Proteins; D015513:Oncogene Proteins; D011518:Proto-Oncogene Proteins; D012097:Repressor Proteins; D000099229:RE1-Silencing Transcription Factor; C438378:BBC3 protein, human; C530104:CCND1 protein, human; C475820:CCNE1 protein, human; C106985:DAXX protein, human; C497325:EGR1 protein, human; C505533:FADD protein, human; C491359:SLC15A4 protein, human; C089495:neuronal Cdk5 activator (p25-p35)
keywords: NRSF; REST; glioblastoma; migration; proliferation