In vitro properties of a newly established medulloblastoma cell line, MCD-1.
Medulloblastomas are poorly differentiated brain tumors believed to arise from primitive pleuripotential stem cells, and tend to express mixed neuronal and glial properties. In the present study, we examined immunohistochemical and neurotransmitter phenotypic properties in a newly established medulloblastoma cell line, MCD-1. MCD-1 cells were immortal, not contact-inhibited, but did not grow in soft agar. Immunohistochemical studies showed positive staining for neurofilament protein (NF), neuron-specific enolase (NSE), synaptophysin, MAP 2, tau, NCAM 180, vimentin, and S-100 protein. The cells expressed specific uptake of glutamate, serotonin, and choline, but not GABA or dopamine. A significant increase in process extension was seen in response to agents that enhance intracellular cyclic AMP, especially 3-isobutyl-1-methylxanthine (IBMX). Process formation induced by IBMX was associated with a decrease in cell proliferation as evidenced by a reduction in numbers of cells incorporating 5-bromo-2-deoxyuridine (BrdU). No increase in process extension was observed following exposure to NGF or retinoic acid. MCD-1 cells were shown to produce transforming growth factor beta (TGF beta), and were immunopositive for mutant p53. Transfection assays with the PG13-Luc reporter plasmid, which contains a p53-responsive enhancer element and a luciferase reporter gene, suggested MCD-1 cells are deficient in wild-type p53 and do not activate p53 on treatment with the anticancer agent adriamycin. The MCD-1 cell line is suggested to represent an abnormally differentiated cell type, which has some properties consistent with a multipotent neuronal phenotype while retaining some properties of immature cells of a glial lineage. The MCD-1 cell line can be used to provide a model of a medulloblastoma cell line that is resistant to growth-controlling and anticancer agents.
Moore
K D
KD
Section on Preclinical Neuroscience, Neuropsychiatry Branch, NIMH, Neuroscience Center, St. Elizabeths, Washington, DC 20032, USA.
Dillon-Carter
O
O
Conejero
C
C
Poltorak
M
M
Chedid
M
M
Tornatore
C
C
Freed
W J
WJ
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000293:Adolescent; D001973:Bromodeoxyuridine; D003994:Bucladesine / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002528:Cerebellar Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D005576:Colforsin / Q000494:pharmacology; D046508:Culture Techniques / Q000379:methods; D004298:Dopamine / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D008527:Medulloblastoma / Q000378:metabolism / Q000473:pathology* / Q000601:surgery; D009414:Nerve Growth Factors / Q000494:pharmacology; D009419:Nerve Tissue Proteins / Q000032:analysis; D011994:Recombinant Proteins / Q000096:biosynthesis; D012701:Serotonin / Q000378:metabolism; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000096:biosynthesis; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000096:biosynthesis; D005680:gamma-Aminobutyric Acid / Q000378:metabolism
D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; D016212:Transforming Growth Factor beta; D016159:Tumor Suppressor Protein p53; D005576:Colforsin; D012701:Serotonin; D018698:Glutamic Acid; D005680:gamma-Aminobutyric Acid; D003994:Bucladesine; D001973:Bromodeoxyuridine; D015056:1-Methyl-3-isobutylxanthine; D004298:Dopamine