Abstract
The development of genetically marked animal tumour xenografts is an area of ongoing research to enable easier and more reliable testing of cancer therapies. Genetically marked tumour models have a number of advantages over conventional tumour models, including the easy longitudinal monitoring of therapies and the reduced number of animals needed for trials. Several different methods have been used in previous studies to mark tumours genetically, however all have limitations, such as genotoxicity and other artifacts related to the usage of integrating viral vectors. Recently, we have generated an episomally maintained plasmid DNA (pDNA) expression system based on Scaffold/Matrix Attachment Region (S/MAR), which permits long-term luciferase transgene expression in the mouse liver. Here we describe a further usage of this pDNA vector with the human Ubiquitin C promoter to create stably transfected human hepatoma (Huh7) and human Pancreatic Carcinoma (MIA-PaCa2) cell lines, which were delivered into “immune deficient” mice and monitored longitudinally over time using a bioluminometer. Both cell lines revealed sustained episomal long-term luciferase expression and formation of a tumour showing the pathological characteristics of hepatocellular carcinoma (HCC) and pancreatic carcinoma (PaCa), respectively. This is the first demonstration that a pDNA vector can confer sustained episomal luciferase transgene expression in various mouse tumour models and can thus be readily utilised to follow tumour formation without interfering with the cellular genome.
Highlights
Cancer represents one of the greatest health risks worldwide
This study shows for the first time the establishment of genetically marked murine models of pancreatic and hepatocellular carcinomas using a non-viral episomal plasmid vector
The generation of an effective genetically marked murine tumour model for hepatocellular carcinoma (HCC) and pancreatic carcinoma (PaCa) is an important step in this process as it will enable the effects of potential therapeutics to be more and accurately monitored and will enable more reliable data when developing novel anticancer drugs
Summary
Cancer represents one of the greatest health risks worldwide. there is a growing need for developing novel therapeutics and new advances in animal tumour modelling. Many conventional animal tumour models used in the development of anticancer treatments involve injection of human tumour cells into immunocompromised mice [2,3] followed by standard calliper measurements to assess tumour size, usually as an end-point measurement, after the animal has been sacrificed These models are fairly limited and research has been on-going to develop a genetically marked tumour that would enable non-invasive monitoring of the tumour parameters by in vivo imaging based on light emission from luciferaseexpressing cells or fluorescence from GFP-expressing cells [1]. It allows one to monitor the efficacy of therapeutic interventions such as drug, gene or cell therapies more than with conventional models It facilitates tracking of tumour parameters, such as size and development, as well as enables highly sensitive visualisation of early metastasis and the evaluation of minimal residual disease after therapy [4]. It permits the use of sequential measurements to follow tumour size during treatment so that longitudinal studies can be performed to analyse the effects of therapies over time giving more reliable information and reducing the number of experimental animals [5]
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