Effect of bufotalin on the proliferation, migration, invasion, and epithelial-mesenchymal transition of human colorectal cancer HCT116 cells

Abstract

Objective To investigate the effect of different concentrations of bufotalin (BT) on the proliferation, migration, invasion, and epithelial-mesenchymal transition of human colorectal cancer HCT116 cells. Methods CCK-8 assay was used to measure cell viability after 24 and 48 h of BT treatment at different concentrations (0, 10, 20, 40, 80, 160, and 320 nmol/L), and the half-maximal inhibitory concentration (IC50) was calculated. The plate colony formation assay was used to verify the effect of BT treatment at the concentrations of 0, 12.5, and 25.0 nmol/L for 14 days on the colony formation ability of HCT116 cells (established as groups A, B, and C). The wound healing assay and the Transwell assay were used to verify the effect of BT treatment at the concentrations of 0, 25, and 50 nmol/L for 24 h on the migration and invasion abilities of HCT116 cells (established as groups A, C, and D). Western blot was used to measure the protein expression levels of E-cadherin and N-cadherin in HCT116 cells after 24 h of treatment in groups A, C and D. Results CCK-8 assay showed that after HCT116 cells were treated by BT for 24 or 48 h, the inhibitory effect of BT on the proliferation of HCT116 cells increased significantly with the increase in the concentration of BT (F=2 106.00,3 725.00,P<0.05), with an IC50 value of 49.59 nmol/L for 24-hour treatment and 24.10 nmol/L for 48-hour treatment. The plate colony formation assay showed that groups B and C had a significantly lower number of colonies of cells than group A (F=159.30,t=12.40,17.32,P<0.05). The wound healing assay and the Transwell assay showed that compared with group A, groups C and D had significantly lower cell migration rate and number of invading cells (F=120.30,296.80,t=12.71-21.27,P<0.05). Western blot showed that BT significantly upregulated the protein expression level of E-cadherin in HCT116 cells (F=2 736.00,P<0.05), and groups C and D had a significantly higher expression level than group A (t=50.27,72.13,P<0.05); BT significantly downregulated the protein expression level of N-cadherin (F=626.80,P<0.05), and groups C and D had a significantly lower expression level than group A (t=26.54,33.57,P<0.05). Conclusion BT can significantly inhibit the proliferation, migration, invasion, and epithelial-mesenchymal transition of human colorectal cancer HCT116 cells and is expected to become a potential candidate for the treatment of colorectal cancer.

Keywords

• colorectal neoplasm|hct116 cells|bufanolides|cell proliferation|cell movement|neoplasm invasiveness|epithelial-mesenchymal transition