Abstract
Aim:
Epigallocatechin-3-gallate (EGCG) is the major polyphenolic constituent in green tea. The aim of this study is to investigate the effects of EGCG on proliferation and migration of the human colon cancer SW620 cells.
Methods:
Proliferation and migration of SW620 cells were induced by the protease-activated receptor 2-agonist peptide (PAR2-AP, 100 μmol/L) or factor VIIa (10 nmol/L), and analyzed using MTT and Transwell assays, respectively. The cellular cytoskeleton was stained with rhodamine-conjugated phalloidin and examined with a laser scanning confocal fluorescence microscope. The expression of caspase-7, tissue factor (TF) and matrix metalloproteinase (MMP)-9 in the cells was examined using QT-PCR, ELISA and Western blot assays. The activation of extracellular signal-regulated kinase 1 and 2 (ERK1/2) and nuclear factor-kappa B (NF-κB) signaling pathways was analyzed with Western blot.
Results:
Both PAR2-AP and factor VIIa promoted SW620 cell proliferation and migration, and caused cytoskeleton reorganization (increased filopodia and pseudopodia). Pretreatment with EGCG (25, 50, 75, and 100 μg/mL) dose-dependently blocked the cell proliferation and migration induced by PAR2-AP or factor VIIa. EGCG (100 μg/mL) prevented the cytoskeleton changes induced by PAR2-AP or factor VIIa. EGCG (100 μg/mL) counteracted the down-regulation of caspase-7 expression and up-regulation of TF and MMP-9 expression in the cells treated with PAR2-AP or factor VIIa. Furthermore, it blocked the activation of ERK1/2 and NF-κB (p65/RelA) induced by PAR2-AP or factor VIIa.
Conclusion:
EGCG blocks the proliferation and migration of SW620 cells induced by PAR2-AP and factor VIIa via inhibition of the ERK1/2 and NF-κB pathways. The compound may serve as a preventive and therapeutic agent for colon cancers.
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Acknowledgements
This project was supported by the Provincial Science Foundation of Jiangsu (No BK2010336) and the Student's Scientific Research of Jiangsu University (No 09A080).
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Supplementa1 Figure 1
The cell cycle distribution (A) of SW620 cells treated with EGCG or/and VIIa and the percentages of cell cycle phase (B) SW620 cells were plated at 1 × 106 cells in 6-well plates and pretreated with different concentrations of EGCG (0, 25, 50, 75, 100 μg/L) for 15 min, then incubated with VIIa (10 nmol/L) for 8 h. (JPG 186 kb)
Supplementa1 Figure 2
The effects of EGCG on SW620 cell viability by trypan blue exclusion SW620 cells were seeded at 1×104/well into a flat bottom 96-well plate and treated with different concentrations of EGCG (0, 25, 50, 75, 100 μg/L) for 6, 12 and 24 h. (JPG 61 kb)
Supplementa1 Figure 3
The images of migration assay SW620 cells were pretreated with or without EGCG (100 μg/ml) for 15 min and then incubated with PAR2-AP (100 μmol/L) or factor VIIa (10 nmol/L) for 8 h. (JPG 159 kb)
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Zhou, F., Zhou, H., Wang, T. et al. Epigallocatechin-3-gallate inhibits proliferation and migration of human colon cancer SW620 cells in vitro. Acta Pharmacol Sin 33, 120–126 (2012). https://doi.org/10.1038/aps.2011.139
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DOI: https://doi.org/10.1038/aps.2011.139
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