Abstract
CD133 (Prominin-1/AC133) is generally treated as a cell surface marker found on multipotent stem cells and tumor stem-like cells, and its biological function remains debated. Genetically modified rat glioma cell lines were generated by lentiviral gene delivery of human CD133 into rat C6 glioma cells (hCD133(+) -C6) or by infection of C6 cells with control lentivirus (mock-C6). Stable hCD133 expression promoted the self-renewal ability of C6-formed spheres with an increase in the expression of the stemness markers, Bmi-1 and SOX2. Akt phosphorylation, Notch-1 activation, and Notch-1 target gene expression (Hes-1, Hey1 and Hey2) were increased in hCD133(+) -C6 when compared to mock-C6. The inhibition of Akt phosphorylation, Notch-1 activation, and Hes-1 in hCD133(+) -C6 cells effectively suppressed their clonogenic ability, indicating that these factors are involved in expanding the growth of hCD133(+) -C6. An elevated expression of GTPase-activating protein 27 (Arhgap27) was detected in hCD133(+) -C6. A decline in the invasion of hCD133(+) -C6 by knockdown of Arhgap27 expression indicated the critical role of Arhgap27 in promoting cell migration of hCD133(+) -C6. In vivo study further showed that hCD133(+) -C6 formed aggressive tumors in vivo compared to mock-C6. Exposure of hCD133(+) -C6 to arsenic trioxide not only reduced Akt phosphorylation, Notch-1 activation and Hes-1 expression in vitro, but also inhibited their tumorigenicity in vivo. The results show that C6 glioma cells with stable hCD133 expression enhanced their stemness properties with increased Notch-1/Hes-1 signaling, Akt activation, and Arhgap27 action, which contribute to increased cell proliferation and migration of hCD133(+) -C6 in vitro, as well as progressive tumor formation in vivo.
Keywords:
Akt; Arhgap27; Hes-1; Notch; arsenic trioxide; glioma.
Copyright © 2013 Wiley Periodicals, Inc.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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AC133 Antigen
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Animals
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Antigens, CD / genetics
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Antigens, CD / metabolism*
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Antineoplastic Agents / pharmacology
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Antineoplastic Agents / therapeutic use
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Arsenic Trioxide
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Arsenicals / pharmacology
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Basic Helix-Loop-Helix Transcription Factors / genetics
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Basic Helix-Loop-Helix Transcription Factors / metabolism
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Brain Neoplasms / drug therapy
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Brain Neoplasms / metabolism*
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Brain Neoplasms / pathology
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Cell Line, Tumor
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Cell Movement / drug effects
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Cell Movement / genetics
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Cell Proliferation* / drug effects
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Cell Transformation, Neoplastic / genetics
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Cell Transformation, Neoplastic / metabolism*
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Cell Transformation, Neoplastic / pathology
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Cerebral Cortex / pathology
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Disease Models, Animal
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Dose-Response Relationship, Drug
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Enzyme Inhibitors / pharmacology
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Female
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Formazans
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GTPase-Activating Proteins / genetics
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GTPase-Activating Proteins / metabolism
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Gene Expression Regulation, Neoplastic / drug effects
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Gene Expression Regulation, Neoplastic / genetics
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Glioma / drug therapy
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Glioma / metabolism*
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Glioma / pathology
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Glycoproteins / genetics
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Glycoproteins / metabolism*
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Homeodomain Proteins / genetics
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Homeodomain Proteins / metabolism
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Humans
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Intercellular Signaling Peptides and Proteins / pharmacology
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Lentivirus / genetics
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Oncogene Protein v-akt / genetics
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Oncogene Protein v-akt / metabolism
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Oxides / pharmacology
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Peptides / genetics
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Peptides / metabolism*
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RNA, Messenger / metabolism
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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Rats
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Rats, Sprague-Dawley
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Receptor, Notch1 / genetics
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Receptor, Notch1 / metabolism
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Tetrazolium Salts
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Time Factors
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Transcription Factor HES-1
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Transfection
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Tumor Stem Cell Assay
Substances
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AC133 Antigen
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ARHGAP27 protein, human
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Antigens, CD
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Antineoplastic Agents
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Arsenicals
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Basic Helix-Loop-Helix Transcription Factors
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Enzyme Inhibitors
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Formazans
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GTPase-Activating Proteins
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Glycoproteins
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Hes1 protein, rat
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Homeodomain Proteins
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Intercellular Signaling Peptides and Proteins
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Oxides
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PROM1 protein, human
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Peptides
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Prom1 protein, rat
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RNA, Messenger
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RNA, Small Interfering
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Receptor, Notch1
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Tetrazolium Salts
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Transcription Factor HES-1
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MTT formazan
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Oncogene Protein v-akt
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Arsenic Trioxide