Proteomic study reveals a co-occurrence of gallic acid-induced apoptosis and glycolysis in B16F10 melanoma cells.
Gallic acid (GA) has long been associated with a wide range of biological activities. In this study, its antitumor effect against B16F10 melanoma cells was demonstrated by MTT assay, cell migration assay, wound-healing assay, and flow cytometric analysis. GA with a concentration >200 μM shows apoptotic activity toward B16F10 cells. According to Western blotting data, overexpressions of cleaved forms of caspase-9, caspase-3, and PARP-1 and pro-apoptotic Bax and Bad, accompanied by underexpressed anti-apoptotic Bcl-2 and Bcl-xL indicate that GA induces B16F10 cell apoptosis via mitochondrial pathway. The 2-DE based comparative proteomics was further employed in B16F10 cells with and without GA treatment for a large-scale protein expression profiling. A total of 41 differential protein spots were quantified, and their identities were characterized using LC-MS/MS analysis and database matching. In addition to some regulated proteins that were associated with apoptosis, interestingly, some identified proteins involved in glycolysis such as glucokinase, α-enolase, aldolase, pyruvate kinase, and GAPDH were simultaneously up-regulated, which reveals that the GA-induced cellular apoptosis in B16 melanoma cells is associated with metabolic glycolysis.
Hyperbaric Oxygen Therapy Center and Division of Plastic Surgery, Chi Mei Medical Center , Tainan 710, Taiwan.
Cheng
C
Liu
Jen-Jie
JJ
Lin
Zih-Yan
ZY
Yang
Chi-Chu
CC
Tsai
Jue-Liang
JL
Hsu
Yu-Jen
YJ
Wu
D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D005707:Gallic Acid / Q000494:pharmacology*; D006019:Glycolysis / Q000187:drug effects; D006801:Humans; D008545:Melanoma / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000503:physiopathology*; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D040901:Proteomics; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism
D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; D005707:Gallic Acid; D053148:Caspase 3
2-DE; apoptosis; gallic acid; glycolysis; mitochondrial dysfunction; proteomics