Extract from Modified Xiao Xianxiongtang Inhibits Epithelial-mesenchymal Transition and Invasion and Migration Mediated by TGF-β1 of Human Gastric Cancer MGC-803 Cells via Wnt5a/Ca2+/NFAT Signaling Pathway
10.13422/j.cnki.syfjx.20202326
- VernacularTitle:加味小陷胸汤水提物通过Wnt5a/Ca2+/NFAT信号通路抑制TGF-β1介导的人胃癌MGC-803细胞上皮-间质转化及侵袭迁移
- Author:
Rui DING
1
;
Rui-rui GE
1
;
En-yu WANG
1
;
Xing-hui HONG
1
;
Liang WANG
1
;
Peng ZHOU
1
;
Jing SHAO
1
;
Hai-yan FANG
1
;
Hui SHI
1
;
Jing-hui WANG
1
;
Jin-ling HUANG
1
Author Information
1. Anhui University of Chinese Medicine，Hefei 230012，China
- Publication Type:Research Article
- Keywords:
aqueous extract from modified Xiao Xianxiongtang;
human gastric cancer MGC-803 cells;
epithelial-mesenchymal transition;
invasion and migration;
Wnt5a/Ca2+/NFAT signaling pathway
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(4):37-46
- CountryChina
- Language:Chinese
-
Abstract:
Objective:This studu aims to investigate the effect of aqueous extract of modified Xiao Xianxiongtang on the epithelial mesenchymal transition（EMT） and the change of its invasion and migration ability of human gastric cancer MGC-803 cells mediated by transforming growth factor-β1（TGF-β1），and to explore the mechanism of regulating Wnt5a/Ca2+/ activated T-cell nuclear factor（NFAT） signaling pathway to inhibit EMT and invasion and metastasis of MGC-803 cells. Method:TGF-β1（10 μg·L-1）was used to induce EMT and the invasion and metastasis model of human gastric cancer MGC-803 cells. Transwell chamber experiment， scratchhealing experiment， Western blot and immunofluorescence assay were used to detect cell invasion and migration ability， expression of EMT marker protein and key protein expression of Wnt5a/Ca2+/NFAT signaling pathway， and intracellular Ca2+ concentration. Result:Compared with the blank group， TGF-β1 could significantly enhance the invasion and migration ability of MGC-803 cells（P<0.01）， down-regulate the level of E-cadherin（P<0.01）， up-regulate protein expressions of N-cadherin， Snail and Vimentin（P<0.01）， and induce cell Wnt5a， calcineurin （CaN）， total protein of activated T-cell nuclear factor 1（NFAT1）， up-regulation of phosphorylated proteins p-NFAT1 and NFAT1 nucleoprotein and intracellular accumulation of Ca2+（P<0.01）. Compared with the TGF-β1 group， modified Xiao Xianxiongtang （10， 20， 40 mg·L-1） could significantly inhibit this phenomenon，and 40 mg·L-1 had the best effect（P<0.05，P<0.01）.The specific inhibitors of Wnt5a/Ca2+/NFAT signaling pathway （R）-（+）-Bay-K-8644 and modified Xiao Xianxiongtang （40 mg·L-1） could significantly inhibit theinvasion and migration of MGC-803 cells mediated by TGF-β1， up-regulate the level of E-cadherin， and down-regulate expressions of N-cadherin， Snail， Vimentin， Wnt5a， CaN and NFAT1 proteins and reduce the intracellular accumulation of Ca2+（P<0.05，P<0.01）.Moreover， （R）-（+）-Bay-K-8644 combined with modified Xiao Xianxiongtang （40 mg·L-1） had stronger inhibitory effect（P<0.05，P<0.01）. Conclusion:These results suggest that modified Xiao Xianxiongtang can inhibit the EMT mediated by TGF-β1 via Wnt5a/Ca2+/NFAT signaling pathway，thereby reducing the invasion and migration ability of MGC-803 cells.
