Mechanism analysis of transgene expression induced by chitosan/hyaluronic acid nanoparticles in CD44v9/RHAMM co-expressed gastric cancer cells: contribution of hyaluronic acid to RHAMM-mediated intracellular transport.

Optimizing non-viral vectors for gene therapy requires elucidating the molecular mechanisms underlying vector uptake and intracellular delivery. In this study, cell transfection using pDNA/polysaccharide complexes was investigated to develop a gene delivery system specific for gastric tumor cells that express hyaluronic acid (HA) receptors such as CD44 variant 9 (CD44v9) and receptor for hyaluronan-mediated motility (RHAMM). pDNA/chitosan/HA complexes showed specific transgene expression activity in CD44v9(+) cells, mediated by CD44v9-dependent cell uptake and microtubule-dependent transport. Particularly, RHAMM was involved in intracellular transport of pDNA/chitosan/HA complexes. We demonstrated that HA drives CD44v9-mediated internalization, RHAMM-mediated, and microtubule-dependent intracellular transport, which are essential for enhanced pDNA complex-driven gene expression in gastric cancer cells co-expressing CD44v9 and RHAMM.