Nuclear accumulation of exogenous DNA fragments in viable cells mediated by FGF-2 and DNA release upon cellular injury

We and others have previously shown that basic fibroblast growth factor (FG F-2 or bFGF) can be used as a targeting molecule to help carry plasmid DNA into cells when the growth factor molecule is physically coupled to the DNA molecule being delivered. Herein we report our observations on the FG;F-me diated uptake of exogenous labeled DNA into cultured cells in a manner that is representative of that which may occur under physiological conditions a t sites of wounded tissue. Cellular debris at such sites contains nucleic a cid fragments released from dead cells, as well as growth factors such as F GF-Q that function early in the wound repair process. Using a cell culture model designed to mimic the local environment of a wound with respect to th e presence of soluble FGF-S and DNA fragments, we have shown that FGF-2 is able to direct the cellular uptake and nuclear localization of fragments of exogenous DNA via the FGF receptor into intact and healthy cells. Furtherm ore, we can monitor and quantitate this type of FGF-mediated DNA delivery b y using indirect immunofluorescence of bromodeoxyuridine-labeled exogenous DNA. Our results suggest that this type of FGF-mediated DNA fragment uptake could allow for the transduction of viable nearest neighbor cells at sites of injury in vivo. Such a phenomenon may lead to mutational aberrations in the recipient cells and enhance the probability of wound carcinogenesis. ( C) 2001 Academic Press.