Fig. 5. CME regulates apoptosis-mediated proteins through reducing COX-2 expression by regulating the Akt/GSK-3β/mTOR and Akt/GSK-3β/β-catenin signaling pathways in HT-29 colon cancer cells. (a) Proteins expression of p-mTOR, mTOR, p-TSC2, TSC2, p-β-catenin, β-catenin, COX-2, p-Akt, Akt, p-GSK-3β, GSK-3β, procaspase-3, Bcl-2, Bak, and Bax. (b) Cells were treated with 20 μM Celecoxib or 1 μM XAV939 or 60 μg/mL CME for 24 h. Protein levels were determined by Western blot analysis. The β-actin probe served as protein-loading control. (c) Caspase-3 activity assay. The statistical analysis of the data was carried out by use of independent sample t-test. **p < 0.01 and ***p < 0.001 vs. con. ###p < 0.001 vs. CME-treated group (each experiment, n = 3). (d) CME, Celecoxib and XAV939 effects on COX-2 mRNA expression. Representative reverse-transcriptase-PCR pictures are shown with β-actin as the housekeeping gene. The statistical analysis of the data was carried out by use of independent sample t-test. ***p < 0.001 vs. con (each experiment, n = 3). N.S., not significant; CME, Cnidium monnieri (L.) Cusson extract; p, phosphorylated; mTOR, mammalian target of rapamycin; TSC2, tuberous sclerosis complex 2; PARP, Poly (ADP-ribose) polymerase; COX-2, cyclooxygenase-2; Akt, protein kinase B; GSK-3β, glycogen synthase kinase-3β; Bcl-2, B-cell lymphoma 2; Bak, Bcl-2-homologous antagonist killer; Bax; Bcl-2-associated X protein; con, control.
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