Comprehensive characterization of neuroblastoma cell line subtypes reveals bilineage potential similar to neural crest stem cells.
BACKGROUND
Neuroblastic tumors (NBT) derive from neural crest stem cells (NCSC). Histologically, NBT are composed by neuroblasts and Schwannian cells. In culture, neuroblastic (N-), substrate-adherent (S-) and intermediate phenotype (I-) cell subtypes arise spontaneously.


METHODS
Here, neuroblastoma (NB) cell line subtypes were characterized according to embryonic peripheral nervous system development markers (GAP43, Phox2b, Sox10, c-kit, GD2, NF68, vimentin, S100beta, calcyclin and ABCG2), morphological features, gene expression and differentiation potential. I-type cells were investigated as a bipotential (neuronal and glial) differentiation stage.


RESULTS
Positive immunostaining of NCSC (GAP43, c-kit, NF68, vimentin and Phox2b) and undifferentiated cell (ABCG2) markers was observed in all NB subtypes. N- and I-type cells displayed cytoplasmic membrane GD2 staining, while nuclear calcyclin was restricted to S-type. N- and I-type cells showed similar phenotype and immunoreactivity pattern. Differential gene expression was associated with each cell subtype. N- and I-type cells displayed similar differentiation capacity towards neuronal and glial lineage fates. S-type cells, upon induction, did not show a neuronal-like phenotype, despite gene expression changes.


CONCLUSIONS
Results suggest that N- and I-type NB cell subtypes represent an immature bilineage stage, able to progress towards neuronal and glial fates upon induction of differentiation. S-type cells appear irreversibly committed to a glial lineage fate.
Acosta
Sandra
S
Developmental tumor biology laboratory, Hospital Sant Joan de Déu, Barcelona, Spain. sacosta@fsjd.org
Lavarino
Cinzia
C
Paris
Raquel
R
Garcia
Idoia
I
de Torres
Carmen
C
Rodríguez
Eva
E
Beleta
Helena
H
Mora
Jaume
J
D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D001973:Bromodeoxyuridine / Q000494:pharmacology; D018797:Cell Cycle Proteins / Q000235:genetics; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor; D019070:Cell Lineage / Q000235:genetics / Q000502:physiology*; D019922:GAP-43 Protein / Q000378:metabolism; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D017350:N-Acetylgalactosaminyltransferases / Q000235:genetics; D009363:Neoplasm Proteins / Q000378:metabolism; D009432:Neural Crest / Q000166:cytology*; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology*; D016900:Neurofilament Proteins / Q000378:metabolism; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins / Q000235:genetics; D055757:SOXE Transcription Factors / Q000235:genetics; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology; D014746:Vimentin / Q000378:metabolism
C116614:ABCG2 protein, human; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D018797:Cell Cycle Proteins; D019922:GAP-43 Protein; D018398:Homeodomain Proteins; C109220:NBPhox protein; D009363:Neoplasm Proteins; D016900:Neurofilament Proteins; D000075383:S100 Calcium Binding Protein A6; D009418:S100 Proteins; C527923:SOX10 protein, human; D055757:SOXE Transcription Factors; D014157:Transcription Factors; D014746:Vimentin; C044882:neurofilament protein L; C055351:S100A6 protein, human; D014212:Tretinoin; D017350:N-Acetylgalactosaminyltransferases; C023220:(N-acetylneuraminyl)-galactosylglucosylceramide N-acetylgalactosaminyltransferase; D019009:Proto-Oncogene Proteins c-kit; D001973:Bromodeoxyuridine