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6-Shogaol Inhibits the Cell Migration of Colon Cancer by Suppressing the EMT Process Through the IKKβ/NF-κB/Snail
Min Chen1,2, Chiin Tong1, Qibiao Wu1,3,4
1Faculty of Chinese Medicine and State Key Laboratory of Quality Research in Chinese Medicines, Macau University of Science and Technology, Macau, Macau SAR,, China.
Abstract:
6-Shogaol from ginger has anti-inflammatory, anti-oxidation and anti-cancer effects. Aim of the Study: To study the effects and possible mechanisms of 6-Shogaol on inhibiting the migration of colon cancer cells Caco2 and HCT116 and prove the effects on proliferation and apoptosis. Materials and methods: The cells were treated with 6-Shogaol at the concentrations of 20, 40, 60, 80, and 100 µM, the cytotoxicity was tested by Colony formation assays and 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and the Western blot was used to evaluate IKKβ/NF-κB/Snail pathway and EMT-related proteins. In addition, in order to eliminate the interference of proliferation inhibition on the experiment, Caco2 cells were treated with 6-Shogaol at the concentrations of 0, 40, and 80 µM, HCT116 cells were treated with 6-Shogaol at the concentrations of 0, 20, and 40 µM, apoptosis was measured by Annex V/PI staining, and migration was measured by Wound healing assays and Transwell test. Results: 6-Shogaol significantly inhibited the growth of cells. The maximum inhibitory concentration of half of them was 86.63 µM in Caco2 cells and 45.25 µM in HCT116 cells. At 80 µM and 40 µM concentrations, 6-Shogaol significantly promoted apoptosis of colon cancer Caco2 cells and HCT116 cells, and also significantly inhibited cell migration (P < .05). In addition, Western blot analysis showed that at 80 µM dose of 6-Shogaol significantly reduced MMP-2, N-cadherin, IKKβ, P-NF-κB and Snail expression in Caco2 cells (P < .05). 40 µM dose of 6-Shogaol significantly reduced VEGF, IKKβ, and P-NF-κB expression, and MMP-2, N-cadherin and Snail was significantly decreased at 60 µM of 6-Shogaol in HCT116 cells(P < .05). However, there was no significant change in E-cadherin in Caco2 cells, and the expression of E-cadherin protein in HCT116 cells decreased. Conclusion: This study proposes and confirms that 6-Shogaol can significantly inhibit the migration of colon cancer cells Caco2 and HCT116, and its mechanism may be produced by inhibiting EMT through IKKβ/NF-κB/Snail signaling pathway. It was also confirmed that 6-Shogaol inhibited the proliferation and promoted apoptosis of Caco2 and HCT116 cells.
Insights
6-Shogaol from ginger effectively inhibits colon cancer cell migration and proliferation while promoting apoptosis. Its mechanism involves inhibiting epithelial-mesenchymal transition (EMT) via the IKKβ/NF-κB/Snail pathway.
Area of Science:
- Oncology
- Pharmacology
- Molecular Biology
Background:
- 6-Shogaol, a compound found in ginger, exhibits anti-inflammatory, antioxidant, and anti-cancer properties.
- Colon cancer cell migration is a critical factor in metastasis and patient prognosis.
Purpose of the Study:
- To investigate the effects of 6-Shogaol on colon cancer cell migration, proliferation, and apoptosis.
- To elucidate the underlying molecular mechanisms, focusing on the IKKβ/NF-κB/Snail signaling pathway and epithelial-mesenchymal transition (EMT).
Main Methods:
- Colon cancer cell lines (Caco2 and HCT116) were treated with varying concentrations of 6-Shogaol.
- Cytotoxicity was assessed using MTT and colony formation assays.
- Apoptosis, cell migration, and protein expression (IKKβ/NF-κB/Snail pathway, EMT markers) were evaluated using Annex V/PI staining, wound healing assays, Transwell tests, and Western blotting.
Main Results:
- 6-Shogaol significantly inhibited the proliferation of Caco2 and HCT116 cells.
- The compound promoted apoptosis in both cell lines.
- 6-Shogaol markedly reduced cell migration and inhibited key EMT markers, including MMP-2, N-cadherin, IKKβ, P-NF-κB, and Snail, while affecting E-cadherin expression differently in the two cell lines.
Conclusions:
- 6-Shogaol demonstrates significant potential in inhibiting colon cancer cell migration and proliferation.
- The anti-migratory effects are likely mediated by the inhibition of EMT through the IKKβ/NF-κB/Snail signaling pathway.
- 6-Shogaol also promotes apoptosis in colon cancer cells, suggesting its therapeutic value.
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