逆萎康含药血清调控AKT/GSK-3β/β-catenin通路抑制MC细胞的上皮间质转化

Serum containing Niweikang inhibits epithelialmesenchymal transition of MC cells via AKT/GSK-3β/β-catenin signaling pathway

目的:逆萎康(NWK)含药血清对MC细胞(MNNG诱导人胃黏膜上皮细胞系(GES-1)恶性转化)上皮间质转化(EMT)的影响及可能机制。方法:用不同浓度含药血清作用于MC细胞,采用蛋白免疫印迹法(Western blotting)检测E-cadherin、N-cadherin、vimentin、p-AKT Ser473、β-catenin、α-SMA、GSK-3β、p-GSK-3βSer9、AKT蛋白的表达影响;采用实时荧光定量聚合酶链式反应(Real-time PCR)检测C-myc基因的表达影响。结果:与空白组比较,蛋白质印迹实验表明,逆萎康含药血清、AKT抑制剂GSK690693(10μmol·L-1)使得β-catenin、p-AKT Ser473、E-cadherin和p-GSK-3βSer9在蛋白水平的表达下调(P<0.05),而α-SMA、N-cadherin和vimentin在蛋白水平上的表达上调(P<0.05),但GSK-3β和AKT这两种蛋白的表达差异不显著;qRT-PCR结果证实逆萎康含药血清可以抑制C-myc mRNA(P<0.05)的表达,并且加入抑制剂后抑制作用增强,差异具有显著性。结论:逆萎康通过抑制AKT/GSK-3β/β-catenin信号通路的活化,抑制MC细胞的EMT过程。

OBJECTIVE To investigate the effect of Niweikang-containing serum on epithelial mesenchymal transition(EMT)of MC cells(MNNG-induced malignant transformation of human gastric epithelial cell line GES-1)and its possible mechanism.METHODS MC cells were treated with different concentrations of drug-containing serum.The expression of E-cadherin,N-cadherin,vimentin,p-AKT Ser473,β-catenin,α-SMA,GSK-3β,p-GSK-3βSer9and AKT proteins was detected by Western blot,and the expression of C-myc gene was detected by real-time quantitative polymerase chain reaction(Real-time PCR).RESULTS Compared with the blank group,Western blotting showed that the expression ofβ-catenin,p-AKT Ser473,E-cadherin and p-GSK-3βSer9 were down-regulated(P<0.05),while the expression ofα-SMA,N-cadherin and vimentin were up-regulated(P<0.05).However,there was no significant difference in the expression of GSK-3βand AKT.The results of qRT-PCR confirmed that the Niweikang-containing serum could inhibit the expression of C-myc mRNA(P<0.05),and the inhibition was enhanced after adding the inhibitor and the difference was significant.CONCLUSION Niweikang inhibits the EMT process of MC cells by inhibiting the activation of AKT/GSK-3β/β-catenin signaling pathway.