Establishment and Characterization of FusionRed Stable Transfected Canine Prostate Adenocarcinoma and Transitional Cell Carcinoma Cells

In Vivo. 2022 Jan-Feb;36(1):170-179. doi: 10.21873/invivo.12688.

Abstract

Background/aim: Cancer cell inoculation is routinely used to evaluate novel therapeutic approaches in vivo. However, without reporter genes enabling deep tissue imaging, study of early tumor progression and therapeutic responses is often limited. We describe the establishment and characterization of two canine cancer cell lines stably expressing red fluorescence proteins as tools for later in vivo imaging.

Materials and methods: Two red fluorescence cell lines were generated by plasmid transfection. Fluorescence protein expression was confirmed by flow cytometry and microscopy. Deep tissue imaging was demonstrated in mice using a NightOWL LB 983. Gene expression changes after transfection were analyzed by RNAseq.

Results: Both cell lines were detectable in vivo by subcutaneous injection of 1×106 cells. RNAseq revealed up to 2005 transfection-induced differentially expressed genes but no significant changes in cellular key pathways.

Conclusion: The fluorescent cell lines provide a solid basis for future in vivo studies on canine cancer.

Keywords: Prostate cancer; in vitro and in vivo imaging; red fluorescent protein.

MeSH terms

  • Adenocarcinoma* / genetics
  • Animals
  • Carcinoma, Transitional Cell*
  • Cell Line, Tumor
  • Dogs
  • Humans
  • Male
  • Mice
  • Prostate
  • Prostatic Neoplasms* / genetics
  • Transfection