灯盏花素通过PI3K/AKT轴抑制胃腺癌细胞增殖与侵袭OA

Objective:To evaluate the inhibitory effects of breviscapine—the major active component derived from the traditional Chinese medicinal herb Erigeron breviscapus—on the proliferation,migration,and invasion of AGS gastric adenocarcinoma cells,and to elucidate its underlying molecular mechanisms.Methods:AGS cells were treated with different concentrations of breviscapine(0-50 μmol/L).Cell proliferation was assessed using CCK-8 assays,colony formation assays,and EdU staining.Cell migration and invasion were evaluated through wound-healing and Transwell assays.Flow cytometry was performed to analyze cell cycle distribution and apoptosis.Western blot was used to examine the expression of proliferation-related proteins(PCNA,Cyclin D1,MYC),epithelial-mesenchymal transition(EMT)markers(E-CAD,N-CAD,MMP2),and key molecules in the PI3K/AKT signaling pathway.Results:CCK-8 assays showed that the viability of AGS cells was significantly reduced in a concentration-dependent manner after treatment with 20～50 μmol/L breviscapine for 24 h and 48 h(P<0.01),while breviscapine at all tested concentrations had no significant effect on the viability of normal gastric mucosal GES-1 cells.Colony formation assays and EdU staining revealed that the number of colony formation and the proportion of proliferating cells in the S phase were significantly decreased in AGS cells treated with 20 μmol/L and 30 μmol/L breviscapine(P<0.05).Wound-healing and Transwell assays demonstrated that 20 μmol/L and 30 μmol/L breviscapine significantly decreased the wound healing rate,the number of migrated and invaded AGS cells(P<0.01),with the inhibitory effect enhanced as the concentration increased.Flow cytometry analysis indicated that the proportion of AGS cells in the G1 phase was significantly increased and that in the S phase was signifi‐cantly decreased after treatment with 20 μmol/L and 30 μmol/L breviscapine(P<0.01);the apoptosis rate of AGS cells in the 30 μmol/L breviscapine group reached 14.3%,which was significantly higher than 1.93%in the control group(P<0.01).Western blot results showed that 20 μmol/L and 30 μmol/L breviscapine significantly downregulated the expression of proliferation-related proteins including PCNA,Cyclin D1 and MYC in AGS cells(P<0.05),upregulated the expression of the epithelial marker E-CAD,and downregulated the expression of the mesenchymal marker N-CAD and invasion-related protein MMP2(P<0.01).Moreover,breviscapine significantly upregulated the expression of pro-apoptotic proteins BAX and C-Caspase-3,downregulated the expression of anti-apoptotic protein Bcl-2(P<0.01),and markedly decreased the phosphorylation levels of PI3K and AKT(P<0.01).After combined treatment with a PI3K/AKT pathway activator,the expression levels of p-PI3K and p-AKT in AGS cells were significantly increased compared with the breviscapine alone group(P<0.01),and the inhibitory effects of breviscapine on the proliferation,migration and invasion of AGS cells were significantly attenuated(P<0.01).Conclusion:Breviscapine effectively suppresses the proliferation,migration,and invasion of AGS gastric adenocarcinoma cells by inhibiting the PI3K/AKT signaling pathway and modulating proliferation-and EMT-related molecules.These findings provide important experimental evidences supporting breviscapine as a potential therapeutic candidate for gastric adenocarcinoma.