Establishment and characterization of a novel uterine carcinosarcoma cell line, TU-ECS-1, with mutations of TP53 and KRAS

Hum Cell. 2017 Apr;30(2):140-148. doi: 10.1007/s13577-016-0154-6. Epub 2016 Nov 26.

Abstract

A new human uterine carcinosarcoma (UCS) cell line, TU-ECS-1, was established and characterized. The morphological appearance of the cultured cells was an insular of epithelial-like cells arranged in the form of a jigsaw puzzle and mesenchymal-like cells with a spindle-shaped or fibroblast-like morphology. A relatively high proliferation rate was observed with a doubling time of 18.2 h. The chromosome number ranged from 44 to 49 and had an extra chromosome 12 (trisomy 12). The respective half-maximal inhibitory concentrations of cisplatin, paclitaxel, and doxorubicin were 2.9 µM, 154 nM, and 219 ng/mL, respectively. Mutational analysis revealed that TU-ECS-1 cells have mutations of TP53 in exons 4, 6, and 8 and of KRAS at codon 12 (G12D) in exon 2, which is a mutation hot spot on this gene. Western blot analysis showed that p53 protein was overexpressed in TU-ECS-1 cells. Immunostaining of the cultured cells and in vivo tumors showed that the TU-ECS-1 cells and xenografts were positive for epithelial marker cytokeratin AE1/3 and mesenchymal marker vimentin. These results suggested that TU-ECS-1 cells might have both epithelial and mesenchymal characteristics. This cell line may be useful to study the carcinogenesis of UCS and contribute to the development of novel treatment strategies.

Keywords: Establishment; KRAS; Molecular-targeted therapy; TP53; Uterine carcinosarcoma.

MeSH terms

  • Animals
  • Carcinosarcoma / drug therapy
  • Carcinosarcoma / genetics*
  • Carcinosarcoma / pathology*
  • Cell Line, Tumor
  • Cell Proliferation / genetics*
  • Cell Separation
  • Cell Transformation, Neoplastic / genetics*
  • Female
  • Humans
  • Mice, Inbred BALB C
  • Molecular Targeted Therapy
  • Mutation*
  • Neoplasm Transplantation
  • Proto-Oncogene Proteins p21(ras) / genetics*
  • Tumor Suppressor Protein p53 / genetics*
  • Uterine Neoplasms / drug therapy
  • Uterine Neoplasms / genetics*
  • Uterine Neoplasms / pathology*

Substances

  • KRAS protein, human
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • Proto-Oncogene Proteins p21(ras)