Establishment of a new cell line with neuronal differentiation derived from small cell neuroendocrine carcinoma of the maxillary sinus

Oncology. 2004;66(3):234-43. doi: 10.1159/000078000.

Abstract

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.

MeSH terms

  • Animals
  • Blotting, Western
  • Bucladesine / pharmacology
  • Carcinoma, Neuroendocrine
  • Carcinoma, Small Cell* / chemistry
  • Cell Adhesion Molecules / analysis
  • Cell Differentiation / drug effects
  • Cell Line, Tumor* / drug effects
  • Chromosome Aberrations
  • DNA, Neoplasm / analysis
  • Gastrin-Releasing Peptide / analysis
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Immunohistochemistry
  • Lymphatic Metastasis
  • Maxillary Sinus Neoplasms* / chemistry
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Neurons / chemistry
  • Neurons / drug effects*
  • Phenotype
  • Phosphopyruvate Hydratase / analysis
  • Polymerase Chain Reaction

Substances

  • Cell Adhesion Molecules
  • DNA, Neoplasm
  • NRCAM protein, human
  • Nrcam protein, mouse
  • Bucladesine
  • Gastrin-Releasing Peptide
  • Phosphopyruvate Hydratase