Di-san junyi daxue xuebao (Aug 2021)
Anti-tumor mechanism of evodiamine on gastric cancer cells in vitro through MAPK signaling pathway
Abstract
Objective To investigate the anti-tumor effect of evodiamine on gastric cancer cells via the MAPK signaling pathway. Methods MTT assay was used to assess the cell viability of SGC-7901 cells after the treatment of 8, 16 and 32 μmol/L evocarpine (denoted as A1 group, A2 group, A3 group). The effect of p38 inhibitor on the above cells (denoted as B1 group, B2 group, B3 group) were studied. The blank control cell group was the cells treated with culture medium. Annexin V-FITC/PI was used to observe cell apoptosis, PI staining to observe cell cycle, Western blotting to measure apoptosis-related proteins Bcl-2 and cleaved-caspase-3, and scratch healing test to evaluate the inhibitory effect on cell migration. Western blotting was employed to detect the expression levels of c-dc2, cdc25C, cyclin B1, and p-ERK and p-p38. Results Evodiamine negatively inhibited the proliferation of SGC-7901 cells in a dose-dependent and time-dependent manner. The IC50 values at 24 and 48 h were 16.78 and 7.28 μmol/L, respectively. Compared with the control group (0.06%), the apoptotic rate of SGC-7901 cells was increased in groups A1, A2 and A3 (18.72%, 31.08% and 52.17%), and the cells arrested at G2/M phase. The proportion of apoptotic cells in group B was decreased, there were less cells arrested at G2/M phase, and the expression of G2/M phase regulatory proteins CDC2, CDC25C and Cyclin B1 were down-regulated when compared with group A (all P < 0.05). The scratch healing test showed that the healing rate of SGC-7901 cells was decreased with the increase of evodiamine concentration. Compared with groups A1, A2 and A3 (59.13%, 23.15% and 18.31%), the rate was increased in groups B1, B2 and B3 (63.17%, 55.27%, 34.08%, P < 0.05). Western blot results indicated that evodiamine treatment induced the up-regulation of Bcl-2 and cleaved caspase-3, down-regulation of p-ERK, and obvious elevation of p-p38 (P < 0.05). Compared with group A, the expression levels of CDC2, CDC25C and Cyclin B1 were increased, those of Bcl-2 and cleaved caspase-3 were significantly decreased, while that of p-ERK was up-regulated and that of p-p38 was down-regulated in group B (P < 0.05). Conclusion Evodiamine shows anti-tumor effect on gastric cancer cells, and it is realized by the down-regulation of p-p38 protein and up-regulation of apoptosis-related proteins through MAPK signaling pathway.
Keywords
