Visit our Expo - Redox and Inflammation signaling 2012

Session XVII : Cell signaling in health and disease Poster XVII, 5
Sustained gene transfer with photochemical internalisation in breast cancer cells.
Sanae Bouali, Jihane Mirouah, Jean L. Merlin
Laboratoire de recherche en oncologie, EA 3452, Centre Alexis Vautrin, Avenue de
Bourgogne, 54511 Vandoeuvre Les Nancy cedex, France. E-MAIL :
s.bouali@nancy.fnclcc.fr
Although the vectors used to deliver therapeutic genes to targeted cells have traditionally been
modified viruses, non viral synthetic vectors are also receiving much attention as an
alternative method. Most non viral gene therapy vectors deliver transgene into cells through
the endocytic pathway. Lack of escape from endocytic vesicles in many cases constitutes a
barrier for delivery of the transgene. A new biotechnology method was developed named
photochemical internalisation (PCI). PCI has been described as an innovative procedure for
releasing macromolecules from endocytic vesicles to the cytosol of target cells. This
technology is based on light induced delivery of macromolecules such us DNA, proteins, and
other therapeutic molecules in a cytosol.
METHODS: Human breast cancer cells were preincubated with the photosensitiser mesotetraphenylporphine
(TPPS2a) followed by treatment with plasmid encoding enhanced Green
Fluorescent Protein complexed with Polyethylenimine or Eudragit RL or transfectin and light
exposure. The expression of the GFP gene was scored by flow cytometry.
RESULTS: The photochemical using light doses improve the efficiency of transfection
mediated by polyethylenimine , but not by Eudragit RL and transfectin. This study was
designed to optimise the PCI of Polyethylenimine, Eudragit RL and tranfectin-DNA
complexes gene transfer in vitro.
CONCLUSION: In the present study, results confirm the potency of combining the PCI with
Polyethylenimine mediated gene transfer.
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