Abstract
Purpose:
The evidence is increasing that cancer stem cells (CSCs) expressing embryonic and neuronal stem cell markers are present in human retinoblastoma (Rb). This study was conducted to determine whether stem-like cancer cells (SLCCs) in Rb express retinal stem cell-related genes and whether SLCCs can directly differentiate into retinal neurons.
Methods:
The cancer stem cell characteristics in WERI-Rb1 cells were determined with Hoechst 33,342 staining, clone formation assay, and CD133 flow cytometry. The expression of embryonic stem cell and retinal stem cell-related genes was analyzed with real-time PCR and immunofluorescence. The SLCCs were induced to differentiate into retinal neurons by the addition of Dickkopf-related protein 1 and Lefty-A.
Results:
A small but persistent population of cells excluding Hoechst dye in a verapamil-sensitive manner exhibited a cancer stem cell-like phenotype. The SLCCs displayed highly clonogenic abilities and increased CD133 expression with isolation and expansion in culture in serum-free medium. By comparing the expression of stem cell markers, we found Oct3/4 was more highly expressed in the SLCCs than in human embryonic stem cells. Together with the properties of intrinsic retinal stem cell-related gene expression, we found SLCCs can be induced into neuron-like cells that express glial fibrillary acidic protein and rhodopsin (a photoreceptor cell marker).
Conclusions:
These findings provide new insight into cancer stem cells and used a strategy of an artificial change of cancer stem cell fate with transcription factors.
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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Antigens, CD / genetics
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Antigens, CD / metabolism
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Biomarkers / metabolism
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Cell Differentiation / drug effects
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Cell Proliferation / drug effects
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Culture Media, Serum-Free
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Fluorescent Antibody Technique
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Fluorescent Dyes
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Gene Expression / drug effects
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Glial Fibrillary Acidic Protein / genetics
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Glial Fibrillary Acidic Protein / metabolism
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Glycoproteins / genetics
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Glycoproteins / metabolism
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Humans
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Intercellular Signaling Peptides and Proteins / pharmacology
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Left-Right Determination Factors / pharmacology
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Neoplastic Stem Cells / drug effects
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Neoplastic Stem Cells / metabolism
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Neoplastic Stem Cells / pathology*
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Octamer Transcription Factor-3 / genetics
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Octamer Transcription Factor-3 / metabolism
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Organic Cation Transport Proteins / genetics
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Organic Cation Transport Proteins / metabolism
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Peptides / genetics
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Peptides / metabolism
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Real-Time Polymerase Chain Reaction
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Retina / drug effects
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Retina / metabolism
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Retina / pathology*
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Retinal Neurons / drug effects
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Retinal Neurons / metabolism
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Retinal Neurons / pathology*
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Retinoblastoma / metabolism
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Retinoblastoma / pathology*
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Rhodopsin / genetics
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Rhodopsin / metabolism
Substances
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AC133 Antigen
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Antigens, CD
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Biomarkers
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Culture Media, Serum-Free
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DKK1 protein, human
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Fluorescent Dyes
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Glial Fibrillary Acidic Protein
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Glycoproteins
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Intercellular Signaling Peptides and Proteins
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LEFTY2 protein, human
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Left-Right Determination Factors
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Octamer Transcription Factor-3
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Organic Cation Transport Proteins
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POU5F1 protein, human
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PROM1 protein, human
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Peptides
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solute carrier family 22 (organic cation transporter), member 3
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Rhodopsin