Gene therapy strategies have been intensively studied for over 20 years. However, there are yet no gene therapy protocols in conventional use. In this project, a gene delivery protocol was optimized for the transient delivery of mRNA molecules into two human cancer cell lines with the purpose of disclosing novel S100A4 gene interactions as a result of S100A4 overexpression.
The Enhanced Green Fluorescent Protein (EGFP) and the metastasis-promoting S100A4 gene were used in this study. To deliver mRNA molecules we investigated the potential of two different delivery vehicles, the bio-compatible transfection carrier β-cyclodextrin and the well characterized polyethylenimine (PEI) carrier. In addition, we evaluated the possibility for targeted mRNA delivery by the use of photochemical internalization (PCI) technology, a site-specific delivery strategy. Our results showed that PEI was an effective carrier for delivery of mRNA either with or without PCI, which was in contrast to β-cyclodextrin.
To evaluate potential novel S100A4 gene interactions we preformed microarray profiling of two S100A4 mRNA overexpressing cell lines (OHS and LOX). Some of the most affected genes were chosen for reverse transcriptase quantitative PCR verification. The transporter lipocalin 12 and the collagen precursor Col1a2 were confirmed to be affected by S100A4 overexpression.