Analysis of cell adhesion/motility in parotid gland cell lines

The main purpose of our novel technique, chemotactic drug targeting (CDT) is to enhance specific delivery of drugs into target cells via selective induction of their migratory responses. In CDT-ligands professional chemoattractants are preferred, they possess high activity on target cells, while sequences of biologically active substances are valuable as carriers and inducers of internalization. Objectives: to characterize the cell adhesion in human and rat parotid gland cell lines (HSG, Par-C10); to test effects of CDT-conjugates labeled with targeting peptide side chains on adhesion; to analyze the effect of CDT-conjugates on micro-motion; to evaluate anti-proliferative effects of CDT-conjugates as index of potential clinical usability. Methods: Electric Cell-substrate Impedance Sensing (ECIS) technique was applied to measure cell adhesion and micro-motion in a real-time-mode. The tested conjugates were oligo-tuftsin based conjugates substituted with formyl peptide (fMLF) and tuftsin monomers (TKPKG, TKPR) as chemotactic ligands and the drug methotrexate (MTX). Our MATLAB-based routine was used to analyze the micro-motion activity. Results: A significant difference in adhesion characteristics was detected in the two parotid models. The interaction between the surfaces is dependent upon the coating molecules (fibronectin is preferred), while FCS as inducer in the fluid phase is also required. Both tuftsin monomers (TKPKG, TKPR) could decrease the cell adhesion. In T20 CDT-conjugates presence of any side chains (fMLF, TKPKG or TKPR) could result time dependent negative shifts in adhesion, presence of the proliferation inhibitor drug, MTX was not acting in the same manner. Computer based evaluation of micro-motion showed that both coating and the active CDT-ligands can influence this behavior of the cells, however, strong auto/paracrine influences are also conceivable. Conclusions: Cell line specific characteristics were detected in cell adhesion and micro-motion in both model-cells. The adhesion/micro-motion and anti-proliferative effect of tuftsin based CDT-drugs underline the significance of further investigations of these derivatives.