积雪草酸通过Wnt/β-catenin信号通路抑制肺腺癌细胞迁移

Asiatic acid inhibits migration of lung adenocarcinoma cells through Wnt/β-catenin signaling pathway

目的:研究积雪草酸对肺腺癌细胞系H1299及A549细胞迁移的影响及其相关分子机制。方法:用不同浓度积雪草酸(0、20、40、60、80、100μmol/L)处理H1299和A549细胞,采用噻唑蓝(MTT)法检测细胞存活率;流式细胞术检测细胞凋亡情况;细胞划痕实验探究积雪草酸对细胞迁移能力的影响;Western blot检测积雪草酸处理对细胞Wnt/β-catenin信号通路及上皮-间质转化(EMT)相关蛋白表达水平的影响。结果:MTT实验结果显示,随着积雪草酸加药浓度增加和加药时间延长,A549、H1299细胞存活率均下降(P<0.05);流式细胞术检测结果显示积雪草酸可促进细胞凋亡(P<0.05);细胞划痕实验结果表明,积雪草酸抑制H1299及A549细胞迁移(P<0.05);Western blot结果显示,随药物浓度增高,β-catenin、p-GSK-3β^(Ser9)、c-myc、Vimentin、Snail蛋白表达降低,Axin2、E-cadherin蛋白表达增多。结论:在肺腺癌细胞中,积雪草酸抑制EMT的发生,从而抑制肿瘤细胞迁移,其机制可能与抑制Wnt/β-catenin信号通路有关。

Objective:To study the effect of asiatic acid on the migration of lung adenocarcinoma cell lines H1299 and A549 and its related molecular mechanism.Methods:H1299 and A549 cells were treated with different doses of asiatic acid(0,20,40,60,80,100μmol/L),and cell survival rate was measured by MTT assay.Apoptosis was detected by flow cytometry.Cell scratch test was performed to observe the effect of asiatic acid on cell migration.Western blot was used to detect the effects of asiatic acid treatment on Wnt/β-catenin signaling pathway and EMT-related protein expression in cells.Results:MTT assay showed that the survival rates of H1299 and A549 cells were decreased with the increase of asiatic acid concentration and treatment time(P<0.05).Flow cytometry showed that asiatic acid could promote apoptosis(P<0.05).The results of cell scratch test showed that asiatic acid inhibited the migration of H1299 and A549 cells(P<0.05).Western blot results indicated that the expression ofβ-catenin,p-GSK-3β^(Ser9),c-myc,Vimentin and Snail was down-regulated with added drug dose,whereas the expression of Axin2 and E-cadherin was increased.Conclusion:Asiatic acid can check the tumor cell migration by inhibiting epithelial-mesenchymal transition(EMT)in lung adenocarcinoma cells.The mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway.