Establishment and characterization of a VX2 carcinoma derived rabbit cell line as a model system for the study of papilloma virus associated head and neck cancer

Introduction:

For nearly 20 years, the auricular VX2 carcinoma of the New Zealand White (NZW) rabbit served as an animal model for human head and neck squamous cell carcinomas (HNSCC). Similarly as HPV+ HNSCC, the VX2 tumor is the consequence of a papillomavirus (cottontail rabbit papilloma virus, CRPV) infection. Against this background it is of utmost importance to develop a well-characterized VX2 carcinoma derived cell line for in vitro studies. So far, there are only single reports about already established VX2 cell lines referring either to discontinued or not reliably available cell lines.

Methods:

VX2 tissue was obtained from VX2 tumor bearing NZW rabbits and propagated for more than 150 passages in DMEM:F12 (1: 1) media. Validation and characterization of the resulting VX2 carcinoma derived cell line was performed by flow cytometry, fluorescence microscopy, Western blot analysis, qRT-PCR, in situ hybridization and Sanger sequencing.

Results:

High passages (> 150) of VX2 carcinoma derived cells mainly excluded the presence of non-tumor cells and resulted in a highly proliferative cell line. Western blot analysis demonstrated expression of EGFR (epidermal growth factor receptor) in VX2 carcinoma derived cells demonstrating their epithelial origin. qRT-PCR demonstrated detectable transcript levels of CRPV E6 and E7 oncogenes.

Conclusions:

Having a matching VX2 tumor/cell line model system for papillomavirus associated HNSCC will allow to evaluate diagnostic and therapeutic procedures in vitro that otherwise would have been done in vivo, thereby helping to reduce the required number of animal experiments.

Publication History

Publication Date:
23 April 2019 (online)