Transient expression of lysyl oxidase in the lungs of mice

Abstract

Lysyl oxidase is an attractive candidate gene for gene therapy of lung cancer because of its tumour suppressor activity (Contente et al., 1990; Kenyon and Contente, 1991; Tsan et al., 1996) and secretion in the extracellular matrix of the lungs (Kagan et al., 1982). Currently there is little information about the expression and tumour suppressor activity of lysyl oxidase in the lungs. This thesis is focused on this area.

A rabbit uteroglobulin (UG) promoter driven-lysyl oxidase construct (pUGLO) was cloned, then investigated for its transient expression in the lungs of mice using both liposome-mediated gene transfer and naked plasmid DNA gene transfer. To optimise the gene transfer systems, a positive plasmid DNA control containing the UG promoter and the reporter gene chloramphenicolacetyl transferase (CAT). With both the liposome-mediated and naked DNA gene transfer systems, CAT expression was detectable in the lungs of the transfected mice using reverse transcription polymerase chain reaction (RT-PCR). The optimised gene transfer systems were then used for expression studies of lysyl oxidase in the lungs of mice. No expression of lysyl oxidase was detectable.