Rottlerin Inhibits Lonicera japonica-Induced Photokilling in Human Lung Cancer Cells through Cytoskeleton-Related Signaling Cascade
Figure 5
Effects of rottlerin on the photoactivated Lonicera japonica extract-induced disruption of mitochondrial function in CH27 cells. Cells were incubated with vehicle alone or 100 μg/mL Lonicera japonica extracts for 4 h and then irradiated with 0.8 J/cm2 fluence dose. In rottlerin treatment, cells were pretreated with a final concentration of 10 μM rottlerin for 1 h. (a) The fluorescent cation dye JC-1 was used to determine the mitochondrial membrane potential. After irradiation, the cells were harvested and stained with 2 μM JC-1 for 15 min. The mitochondrial depolarization patterns of the CH27 cells were measured by flow cytometry. The sum of the percentage of Q1, Q2, Q3, and Q4 is 100%. (b) The effect of rottlerin on the photoactivated Lonicera japonica extract-induced opening of mitochondrial permeability transition (MPT) pore in CH27 cells. Before treatment with Lonicera japonica extracts and light, cells were loaded with 1 μM calcein AM for 30 min in DMEM medium containing 1 mM CoCl2. After irradiation, the cells were harvested and then analyzed by flow cytometry for loss of fluorescence intensity due to efflux of the dye. (i) control cells; (ii) Lonicera japonica extract-treated cells; (iii) rottlerin-treated cells; (iv) cells were pretreated with rottlerin and then Lonicera japonica extracts. Results are representative of three independent experiments.
