Establishment and characterization of a novel cell line of medullary thyroid carcinoma from a dog

In Vitro Cell Dev Biol Anim. 2019 Aug;55(7):559-566. doi: 10.1007/s11626-019-00339-2. Epub 2019 May 29.

Abstract

A novel cell line of canine medullary thyroid carcinoma (MTC) was established from the neck mass, diagnosed histopathologically and immunohistochemically as ectopic MTC. The neoplastic cells arranging trabecular structures were characterized as pleomorphic cells with eosinophilic cytoplasm and nucleus, containing often clear nucleolus. These tumor cells were immuno-positive for calcitonin gene-related protein (CGRP), somatostatin, and chromogranin A. In addition, 8th passaged cultured cells were also immuno-positive for CGRP, somatostatin, and chromogranin A. The cloned tumor cells showed logarithmic cell growth with a doubling time of 33.3 h. From the results of DNA sequencing of rearranged during transfection (RET) proto-oncogene, the cloned tumor cells had four single base substitution, including exon 5 codon 82, exon 16 codon 750, exon 17 codon 777, and exon 24 codon 1085, all of which were single nucleotide polymorphism reported in RET gene of dogs. After the xenotransplantation into severe combined immunodeficiency (SCID) mice, the cloned cells showed tumorigenicity potentials. The morphological and immunohistochemical features of the xenotransplanted tumor were almost in conformity with those of the original tumor, including positive immunoreactivity for calcitonin, CGRP, and chromogranin A. To our knowledge, this is the first report of canine MTC cell line, which provides useful in vitro tool for understanding oncogenic mechanism and pathophysiological state of MTC in dogs.

Keywords: Cell line; Characteristic; Dog; Medullary thyroid carcinoma; RET gene.

MeSH terms

  • Animals
  • Base Sequence
  • Calcitonin Gene-Related Peptide / metabolism
  • Carcinoma, Neuroendocrine / genetics*
  • Carcinoma, Neuroendocrine / pathology*
  • Cell Culture Techniques
  • Cell Line, Tumor
  • Cell Proliferation
  • Chromogranin A / metabolism
  • Dogs
  • Female
  • Mice
  • Mice, SCID
  • Neoplasm Transplantation
  • Polymorphism, Single Nucleotide / genetics
  • Proto-Oncogene Proteins c-ret / genetics*
  • Sequence Analysis, DNA
  • Somatostatin / metabolism
  • Thyroid Neoplasms / genetics*
  • Thyroid Neoplasms / pathology*
  • Transplantation, Heterologous

Substances

  • Chromogranin A
  • Somatostatin
  • Proto-Oncogene Proteins c-ret
  • Calcitonin Gene-Related Peptide

Supplementary concepts

  • Thyroid cancer, medullary