Differentiation of human retinal pigment epithelial cells into neuronal phenotype by N-(4-hydroxyphenyl)retinamide.
ARPE-19, a human retinal pigment epithelial (RPE) cell line, has been widely used in studies of RPE function as well as gene expression. Here, we report the novel finding that N-(4-hydroxyphenyl)retinamide (fenretinide), a synthetic retinoic acid derivative and a potential chemopreventive agent against cancer, induced the differentiation of ARPE-19 cells into a neuronal phenotype. The treated cells lost their epithelial phenotype and exhibited a typical neuronal shape with long processes (four to five times longer than the cell body). The onset of fenretinide-induced neuronal differentiation was dose and time dependent, started within 1-2 days, and lasted at least 4 weeks. Immunohistochemical studies indicated that the expression of neurofilament proteins (NF160 and NF200), calretinin and neural cell adhesion molecule was increased in these differentiated cells. Western blot analysis indicated that cellular retinaldehyde-binding protein, which is normally expressed in RPE cells, was decreased in treated cells. Protein analysis on a two-dimensional gel followed by matrix-assisted laser desorption ionization-time of flight mass spectrometric analysis demonstrated that heat-shock protein 70 was increased after fenretinide treatment. Thus, fenretinide, a synthetic retinoid, is able to induce neuronal differentiation of human RPE cells in culture.
Chen
Shanyi
S
Biochemistry Section, Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, National Institutes of Health, 6 Center Drive, MSC 2740, Bethesda, MD 20892, USA.
Samuel
William
W
Fariss
Robert N
RN
Duncan
Todd
T
Kutty
R Krishnan
RK
Wiggert
Barbara
B
D000595:Amino Acid Sequence; D016588:Anticarcinogenic Agents / Q000494:pharmacology*; D000943:Antigens, Differentiation / Q000096:biosynthesis; D064032:Calbindin 2; D002352:Carrier Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D017313:Fenretinide / Q000494:pharmacology*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D019006:Neural Cell Adhesion Molecules / Q000378:metabolism; D016900:Neurofilament Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010449:Peptide Mapping; D010857:Pigment Epithelium of Eye / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D064030:S100 Calcium Binding Protein G / Q000378:metabolism; D020539:Sequence Analysis, Protein; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors
C020932:11-cis-retinal-binding protein; D016588:Anticarcinogenic Agents; D000943:Antigens, Differentiation; C576764:CALB2 protein, human; D064032:Calbindin 2; D002352:Carrier Proteins; D018840:HSP70 Heat-Shock Proteins; D019006:Neural Cell Adhesion Molecules; D016900:Neurofilament Proteins; D064030:S100 Calcium Binding Protein G; D017313:Fenretinide