Comparison of pharmacokinetics, intracellular localizations and sonodynamic efficacy of endogenous and exogenous protoporphyrin IX in sarcoma 180 cells.

PURPOSE: The aim of the present study was to investigate the differences in pharmacokinetics, sub-cellular localizations and sonodynamic efficacy between endogenous and exogenous protoporphyrin IX (endo-PpIX and exo-PpIX) in sarcoma 180 (S180) cells. MATERIALS AND METHODS: The 5-aminolevulinic acid (ALA)-derived endo-PpIX and exo-PpIX pharmacokinetic profiles were determined by the fluorescence intensity of cell extracts with a spectrophotometer based on a standard curve. The changes in their sub-cellular localization patterns over a prolonged incubation time were evaluated by laser scanning confocal microscopy. The cytotoxic effects of 5-ALA-mediated sonodynamic therapy (ALA-SDT) and exogenous PpIX-mediated sonodynamic therapy (PpIX-SDT) were also evaluated by the MTT assay. RESULTS: The exo-PpIX showed dose-dependent pharmacokinetics in which a plateau of intra- and extracellular content was observed 45min after administration. However, the amount of ALA-derived endogenous intracellular PpIX, as well as extracellular PpIX in the same samples, showed linear accumulation with incubation time, which was independent of ALA concentration. Fluorescent imaging revealed that the exo-PpIX mainly accumulated at the plasma membrane in the early stage, whereas the ALA-derived PpIX initially localized in the mitochondria. Cells displayed sonodynamic damage by the synthesized endo-PpIX after addition of 1mM ALA for 12h, but the cytotoxicity induced by the equivalent amount of exo-PpIX was much more significant with increasing ultrasound intensities. CONCLUSIONS: Our findings suggest that endo- and exo-PpIX in S180 cells differ not only in pharmacokinetics but also in sub-cellular localizations, which may affect their sonodynamic efficacy and mechanisms of inducing cell death.

Comparisons among sensitivities of different tumor cells to focused ultrasound in vitro.

This study is to test the sensitivities of different tumor cells to ultrasound irradiation at the frequency of 2.2 MHz for 60 s duration, and investigate the potential mechanism underlying different sensitivities. Three murine tumor models with distinct aggressiveness (S180, H-22 and EAC) were exposed to ultrasound to evaluate their sonodynamic efficiencies, and several biological parameters such as cell membrane permeability, lipid peroxidation (LPO), ultra-structure observation, intracellular reactive oxygen species (ROS) and mitochondria membrane potential (MMP) were analyzed after exposures. The results showed that cellular responses of different cells were distinct, of interest to note, the aggressive S180 cells were much more sensitive than others, whereas EAC cells were relatively more resistant to ultrasound irradiation. The direct comparisons among different types of cells indicate that the sono-sensitization seems to depend on the physiological and chemical properties of tumor cells. Perhaps sections of cell membrane became destabilized following the initial radical attack and LPO reaction, which caused S180 cells more susceptible to mechanical stresses during sonolysis. This study provides important implications for cancer therapy.