Establishment of a new cell line with neuronal differentiation derived from small cell neuroendocrine carcinoma of the maxillary sinus.
OBJECTIVE
It is well known that small cell neuroendocrine carcinoma (SNEC) arising at extrapulmonary sites has a poor prognosis and an interesting biological characterization. To understand biological characterization and elucidation of the origin of the histogenesis of SNEC, we report the establishment of a new SNEC cell line and characteristics of neuroendocrine properties including neuronal differentiation by treatment with dibutyryl cyclic AMP (db-cAMP).


METHODS
We established a new cell line (SNEC-MI) derived from SNEC of the maxillary sinus by a modified spill-out method, and verified neuroendocrine properties including neuronal differentiation by immunocytochemical and immunoblotting methods.


RESULTS
The established cell line showed spherical or spindle shape in monolayer culture and was positive for neuron-specific enolase (NSE), neuronal cell adhesion protein (N-CAM, CD56) and gastrin-releasing peptide. NSE was also demonstrated in the cultured medium and dense-core neuroendocrine granules were detected ultrastructurally in the cytoplasm. Treatment of cells with db-cAMP markedly induced the development and elongation of neuronal processes, which formed a netlike arrangement. Characterization of these elongated neuronal processes revealed them immunoreacting intensely with high molecular-weight neurofilament, and a time-dependent increase of microtubule-associated protein-2 in cell lysates.


CONCLUSIONS
These findings indicated that this cell line possesses the capability to differentiate into neuronal cells, and supported the hypothesis that extrapulmonary SNEC might be derived from a pluripotent stem cell.
Noguchi
Kazuma
K
Department of Oral and Maxillofacial Surgery, Hyogo College of Medicine, Nishinomiya, Hyogo, Japan. knoguchi@hyo-med.ac.jp
Urade
Masahiro
M
Kishimoto
Hiromitsu
H
Kuroda
Junko
J
Moridera
Kuniyasu
K
Sakurai
Kazunari
K
D000818:Animals; D015153:Blotting, Western; D003994:Bucladesine / Q000494:pharmacology; D018278:Carcinoma, Neuroendocrine; D018288:Carcinoma, Small Cell / Q000737:chemistry; D015815:Cell Adhesion Molecules / Q000032:analysis; D002454:Cell Differentiation / Q000187:drug effects; D045744:Cell Line, Tumor / Q000187:drug effects; D002869:Chromosome Aberrations; D004273:DNA, Neoplasm / Q000032:analysis; D019886:Gastrin-Releasing Peptide / Q000032:analysis; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007150:Immunohistochemistry; D008207:Lymphatic Metastasis; D008444:Maxillary Sinus Neoplasms / Q000737:chemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009474:Neurons / Q000737:chemistry / Q000187:drug effects*; D010641:Phenotype; D010751:Phosphopyruvate Hydratase / Q000032:analysis; D016133:Polymerase Chain Reaction
D015815:Cell Adhesion Molecules; D004273:DNA, Neoplasm; C491152:NRCAM protein, human; C491151:Nrcam protein, mouse; D003994:Bucladesine; D019886:Gastrin-Releasing Peptide; D010751:Phosphopyruvate Hydratase