土荆皮乙酸通过PI3K/AKT通路抑制肺腺癌细胞A549上皮-间质转化

Epithelial-mesenchymal transition inhibition of lung adenocarcinoma cells A549 by pseudolaric acid B through PI3K/AKT pathway

目的:探讨土荆皮乙酸(PAB)对TGF-β1介导的人肺癌细胞A549上皮-间质转化(EMT)及侵袭能力的影响及作用机制。方法:采用Transwell小室实验检测PAB对A549细胞迁移、侵袭能力的影响。免疫荧光法检测上皮标志蛋白E-钙黏蛋白(E-Cad)表达。Western blot检测上皮相关标志物E-Cad、α-连环蛋白(α-CA)、间质相关标志物波形蛋白(Vim)、α-平滑肌肌动蛋白(α-SMA)以及PI3K、p-PI3K、AKT、p-AKT蛋白表达。RT-qPCR法检测上皮及间质蛋白标志物α-CA、Vim、α-SMA mRNA水平。结果:与空白组相比,诱导组上皮标志蛋白E-Cad、α-CA表达下降,间质标志蛋白Vim、α-SMA表达上调,PI3K/AKT信号通路蛋白PI3K、AKT表达上调,A549细胞迁移、侵袭能力增强。与诱导组相比,实验组与上述结果相反,且A549细胞迁移、侵袭能力减弱。RT-qPCR检测同样验证上述结果。结论:PAB可有效控制A549细胞EMT及迁移、侵袭能力,作用机制可能与调控PI3K/AKT信号通路有关。

Objective:To investigate effect of pseudolaric acid B(PAB)on TGF-β1-mediated human lung cancer cell A549 epithelial-mesenchymal transition(EMT)and invasion ability,and its mechanism.Methods:Transwell was used to detect effect of PAB on migration and invasion of A549 cells. Expression of epithelial marker protein E-cadherin(E-Cad)was detected by immunofluorescence. Epithelial markers E-Cad and α-Catenin(α-CA),stromal associated marker vimentin(Vim)α-Smooth muscle actin(α-SMA),and PI3K,p-PI3K,AKT and p-AKT proteins expressions were detected by Western blot. Epithelial and interstitial protein markers α-CA,Vim,α-SMA mRNA levels were detected by RT-qPCR.Results:Compared with Blank group,expressions of epithelial marker proteins E-Cad,α-CA were decreased in induced group,expressions of mesenchymal marker proteins Vim and α-SMA were was increased,expressions of PI3K/AKT signaling pathway proteins PI3K and AKT were increased,and migration and invasion ability of A549 cells was enhanced. Compared with induced group,experiment group was contrary to the above results and A549 cells migration and invasiveness was down-regulated. RT-qPCR detection also confirmed above results.Conclusion:PAB can effectively control A549 cell EMT and migration and invasion ability,whose mechanism may be related to regulation of PI3K/AKT signaling pathway.