The Cytotoxicity Mechanism of 6-Shogaol-Treated HeLa Human Cervical Cancer Cells Revealed by Label-Free Shotgun Proteomics and Bioinformatics Analysis
Figure 3
Effects of 6-shogaol on expression of proteins on ER stress and mitochondrial apoptosis pathway, as well as the dissipation of ΔΨm on HeLa cells. The levels of PERK, CHOP, ARF5, HSP60 (a), PARP, Pro Caspases-3, and Bax (b). Normalization performed to -actin. The bands shown here were from a representative experiment repeated three times. (c) The caspase 3 activity was detected according to Caspase 3 Activity Assay Kit. Relative caspase-3 activity was calculated as a ratio of emission of 6-shogaol treated cells to vehicle-treated (0.1% DMSO) cells. Values were means ± SD of three independent experiments. ; , compared with control cells. (d) Red fluorescence represents the mitochondrial aggregate form of JC-1, indicating dissipation of ΔΨm. Cells subjected to 15 μM of 6-shogaol were stained by JC-1. Change of ΔΨm was detected by fluorescence microscopy. Vehicle-treated cells which have high ΔΨm show punctuate red fluorescence. Apoptosis cells show diffuse green fluorescence because of decrease in mitochondrial membrane potential.
