Inhibitory Effect of Weichang'an Granule on Invasion and Metastasis of MKN45 Cells in Human Gastric Cancer by RUFY3
10.13422/j.cnki.syfjx.20202464
- VernacularTitle:胃肠安颗粒通过RUFY3抑制人胃癌MKN45细胞侵袭转移
- Author:
Meng-meng QIN
1
;
Jia LI
1
;
Zhao-yan LI
1
;
Ai-guang ZHAO
1
Author Information
1. Longhua Hospital， Shanghai University of Traditional Chinese Medicine（TCM）， Shanghai 200032， China
- Publication Type:Research Article
- Keywords:
Weichang'an granule;
spleen-strengthening traditional Chinese medicine;
gastric cancer;
epithelial mesenchymal transformation（EMT）;
invasion and metastasis;
RUN and FYVE domain protein 3
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2021;27(8):66-73
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the molecular mechanism of spleen-strengthening traditional Chinese medicine （TCM） Weichang'an granule in inhibiting the invasion and metastasis of human gastric cancer MKN45 cells. Method:MKN45 cells were cultured in vitro and incubated with different concentrations（600， 900， 1 200， 1 500 mg·L-1）of Weichang'an granule for 24， 48， 72 h. Cell counting kit-8 （CCK-8） assay was used to detect its effect on the cell proliferation. Western blot was used to detect the expression of RUN and FYVE domain containing 3（RUFY3） in normal gastric mucosa cells and different gastric cancer cell lines. The expression of RUFY3 in the gastric cancer cells after Weichang'an granule intervention （600， 900， 1 200 mg·L-1） was detected by Western blot. Lentivirus transfection technique was used to achieve the stable and silenced expression of RUFY3 in gastric cancer MKN45 cells. Transwell assay was used to evaluate the influence of Weichang'an granule and silenced RUFY3 on the metastasis and invasion ability of MKN45. E-cadherin，N-cadherin，Vimentin，Zinc-finger transcription factor （SNAIL1 and SNAIL2） protein expression levels were detected by Western blot. Result:RUFY3 expression in human gastric cancer cells was significantly higher than that in normal gastric mucosa．The protein expression of RUFY3 in MKN45 cells of silenced RUFY3 group was significantly lower than that in Lentivirus negative group （P<0.01）． Weichang'an granule inhibited the expression of RUFY3 in human MKN45 gastric cancer cells （P<0.05， P<0.01） in a concentration-dependent manner. As compared with the blank group， both Weichang'an granule and silenced RUFY3 inhibited the metastasis and invasion ability of MKN45 （P<0.01）. After Weichang'an granule and silenced RUFY3 treatment， the protein expression of epithelial marker gene E-cadherin was up-regulated （P<0.01）， the protein expression of Vimentin and N-cadherin decreased， but with no statistical difference，while SNAIL1 and SNAIL2 were both significantly down-regulated （P<0.01）. Conclusion:By targeting RUFY3 to regulate epithelial mesenchymal transformation， the spleen-strengthening TCM compound Weichang'an granule can inhibit the invasion and metastasis of gastric cancer.
