Analysis of HepG2 cell response to a wide concentration range of mitomycin C using a multichannel quartz crystal microbalance system with a microscope

Abstract

The morphological response of HepG2 cells to mitomycin C was analyzed using a multichannel quartz crystal microbalance system equipped with a home-built movable microscope that enables the simultaneous acquisition of cell images and measurements of eight-channel quartz crystal microbalance. After 24 h of cell seeding, mitomycin C was injected into the culture medium. During the attachment process, the resonant frequency decreased, and the curves fitted well with the first-order lag response. Analysis of the response to mitomycin C revealed that the resonant frequency response curves varied with mitomycin C concentration. When the mitomycin C concentration was <10 μmol L−1, the delay time was observed before the increase in resonant frequency. When the mitomycin C concentration was extremely low, an additional decrease in resonant frequency was observed in the middle of the delay time that fitted well with the cumulative log-normal distribution curve. The resonant frequency response curves after the delay time fitted well with the cumulative log-normal distribution curves. The delay time and mean cumulative log-normal distribution time for the increase in resonant frequency correlated with the mitomycin C concentration; however, the mean time for the additional decrease in the resonant frequency did not show a statistically significant difference as a function of mitomycin C concentration. For mitomycin C concentrations of >20 μmol L−1, the response to the change in resonant frequency was rapid, and the response curves fitted well with the first-order lag response. The first-order lag response indicates that the response occurred simultaneously for all cells. The results showed that the time constant was independent of the tested mitomycin C concentration between 20 and 100 μmol L−1. These results suggested that different cell death processes occurred by mitomycin C. The findings of this study suggest that the system can be used to investigate cell death in adherent cells.