genetic modification of cancer cells using non-viral, episomal smar vectors for in vivo tumour modelling使用病毒性基因修饰的肿瘤细胞,游离smar向量在体内肿瘤模型.pdfVIP
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genetic modification of cancer cells using non-viral, episomal smar vectors for in vivo tumour modelling使用病毒性基因修饰的肿瘤细胞,游离smar向量在体内肿瘤模型
Genetic Modification of Cancer Cells Using Non-Viral,
Episomal S/MAR Vectors for In Vivo Tumour Modelling
Orestis Argyros., Suet Ping Wong., Kate Gowers, Richard Paul Harbottle*
Gene Therapy Research Group, Section of Molecular Medicine, National Heart and Lung Institute, Imperial College London, London, United Kingdom
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 th
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