Figure 5.

740Y-P reversed the inhibitory effect of CAR on oesophageal cancer cells. (A and B) The oesophageal cancer cell line EC9706 was incubated with CAR and the PI3K agonist 740Y-P or subjected to co-incubation, and Western blot results proved that 740Y-P could partially reverse the inhibitory effect of CAR on the PI3K/AKT pathway. Similarly, EC9706 and TE10 cells were treated with CAR and the PI3K agonist 740Y-P or with co-incubation. (C and D) MTT assay and (E and F) colony formation assay results confirmed that the PI3K agonist 740Y-P could partially reverse the antiproliferation effect of CAR. (G and H) Scratch healing assay, (I and J) Transwell migration assay, and (K and L) Transwell invasion assay results showed that the PI3K agonist 740Y-P can partially reverse the anti-migration and anti-invasion effects of CAR on oesophageal cancer cells. (M and N) Hoechst 33258 staining and (O and P) flow cytometry were used to verify the effect of CAR and 740Y-P or co-incubation on oesophageal cancer cell apoptosis, and the results showed that 740Y-P could partially reverse the apoptosis-promoting effect of CAR on EC9706 and TE10 cells. (Q-V) Western blot and quantitative analysis were used to detect the protein expression levels of Bcl2, Bax, Bad, caspase-3, cleaved caspase-3, MMP2, MMP9, N-cadherin, E-cadherin, Snail, vimentin, and PCNA after EC9706 cells were treated with 740Y-P and CAR or with co-incubation. The results showed that 740Y-P could partially reverse the expression of apoptosis-related proteins, EMT-related proteins, and PCNA. The data are expressed as the mean ± SD (n = 3, each group). Compared with the NC group, *p < 0.05, **p < 0.01, ***p < 0.001; Compared with the 740Y-P group, #p < 0.05, ## p < 0.01, ### p < 0.001; Compared with the Cardamonin group, & p < 0.05, && p < 0.01, &&& p < 0.001.