光甘草定对肺腺癌A549细胞恶性生物学行为的影响及其分子机制

Effects of glabridin on malignant biological behaviors of lung adenocarcinoma A549 cells and its molecular mechanism

目的:探讨光甘草定对肺腺癌细胞A549恶性生物学行的影响及其分子机制。方法:常规方法培养A549细胞和人正常肺上皮细胞BEAS-2B,用不同浓度的光甘草定和或顺铂对其进行处理,通过结晶紫染色、CCK-8法检测光甘草定、顺铂对A549、BEAS-2B细胞增殖活力的影响,Transwell小室实验、细胞划痕实验检测光甘草定对A549细胞侵袭、迁移能力的影响,流式细胞术检测光甘草定对A549细胞凋亡的影响,3D超低黏附板培养法培养A549细胞后采用CCK-8法检测光甘草定对A549细胞增殖的影响,WB法检测光甘草定对A549细胞中上皮间质转化(EMT)相关蛋白表达的影响;构建A549细胞移植瘤模型后检测光甘草定、顺铂对移植瘤的生长以及移植瘤组织中EMT相关蛋白表达的影响。结果:光甘草定、顺铂呈剂量依赖性地显著抑制A549细胞的增殖(P<0.05或P<0.01)、细胞迁移(P<0.05或P<0.01)和侵袭能力(P<0.05或P<0.01),光甘草定能诱导A549细胞凋亡(P<0.01),抑制A549细胞中N-cadherin、snail和vimentin蛋白的表达,促进E-cadherin蛋白表达;光甘草定、顺铂均能抑制A549移植瘤的生长,抑制移植瘤组织中Ki67、N-cadherin、snail和vimentin蛋白的表达、促进E-cadherin蛋白的表达。结论:光甘草定可通过抑制A549细胞的增殖、迁移和侵袭,诱导A549细胞凋亡,其机制可能与其抑制细胞的EMT进程而产生抑癌作用有关。

Objective:To study the effects of glabridin on malignant biological behaviors of lung adenocarcinoma A549 cells and its molecular mechanism.Methods:A549 cells and human normal lung epithelial BEAS-2B cells were cultured by conventional methods and then treated with different concentrations of glabridin and/or cisplatin.The effects of glabridin and cisplatin on the proliferative vitality of A549 and BEAS-2B cells were detected by crystal violet staining and CCK-8 method.Transwell cell assay and cell scratch assay were used to detect the effect of glabridin on the invasion and migration ability of A549 cells.Flow cytometry was conducted to detect the effect of glabridin on the apoptosis of A549 cells.3D ultra-low adhesion plate culture method was applied to culture A549 cells,and then the effects of glabridin on the proliferation of A549 cells and the expression of epithelial mesenchymal transition(EMT)-related proteins in A549 cells were detected using CCK-8 method and Western-blot,respectively.An A549 cell-transplanted tumor model was constructed to detect the effects of glabridin and cisplatin on the growth of transplanted tumors and the expression of EMTrelated proteins in transplanted tumor tissues.Results:Glabridin and cisplatin significantly inhibited the proliferation(P<0.05 or P<0.01),cell migration(P<0.05 or P<0.01)and invasion(P<0.05 or P<0.01)of A549 cells in a dose-dependent manner.Glabridin could induce the apoptosis of A549 cells(P<0.01),inhibit the protein expression of N-cadherin,snail,and vimentin,but promote the protein expression of E-cadherin in A549 cells.Both glabridin and cisplatin could inhibit the growth of A549 cell-transplanted tumors,inhibit the protein expression of Ki67,N-cadherin,snail,and vimentin,and promote the protein expression of E-cadherin in transplanted tumor tissues.Conclusion:Glabridin can inhibit the proliferation,migration,and invasion of A549 cells,induce apoptosis of A549 cells,and inhibit EMT processes,thus exerting anti-tumor effect.