Department of Clinical Laboratory, Jiangsu Province Rongjun Hospital, Wuxi, China.
Acta Pharmacol Sin. 2012 Jan;33(1):120-6. doi: 10.1038/aps.2011.139. Epub 2011 Nov 21.
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.
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.
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.
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.
表没食子儿茶素没食子酸酯（EGCG）是绿茶中主要的多酚类成分。本研究旨在探讨 EGCG 对人结肠癌细胞 SW620 增殖和迁移的影响。
用蛋白酶激活受体 2 激动肽（PAR2-AP，100 μmol/L）或因子 VIIa（10 nmol/L）诱导 SW620 细胞增殖和迁移，分别用 MTT 和 Transwell 检测。用罗丹明标记鬼笔环肽染色细胞骨架，并用激光共聚焦荧光显微镜观察。用 QT-PCR、ELISA 和 Western blot 检测细胞中半胱天冬酶-7、组织因子（TF）和基质金属蛋白酶（MMP）-9的表达。用 Western blot 分析细胞外信号调节激酶 1 和 2（ERK1/2）和核因子-κB（NF-κB）信号通路的激活。
PAR2-AP 和因子 VIIa 均促进 SW620 细胞增殖和迁移，并引起细胞骨架重排（增加丝状伪足和片状伪足）。EGCG（25、50、75 和 100 μg/mL）剂量依赖性地阻断了 PAR2-AP 或因子 VIIa 诱导的细胞增殖和迁移。EGCG（100 μg/mL）阻止了 PAR2-AP 或因子 VIIa 诱导的细胞骨架变化。EGCG（100 μg/mL）对抗了 PAR2-AP 或因子 VIIa 处理细胞中 caspase-7 表达下调和 TF 和 MMP-9 表达上调。此外，它还阻断了 PAR2-AP 或因子 VIIa 诱导的 ERK1/2 和 NF-κB（p65/RelA）的激活。
EGCG 通过抑制 ERK1/2 和 NF-κB 通路阻断 PAR2-AP 和因子 VIIa 诱导的 SW620 细胞增殖和迁移。该化合物可能作为结肠癌的预防和治疗剂。
