基于Wnt/β-catenin信号通路探讨小金丸含药血清抑制乳腺癌细胞的作用机制OA

Objective:To investigate the role and underlying mechanism of the Wnt/β-catenin signaling pathway in the process of Xiaojinwan(XJW)-containing serum interfering with endothelial-mesenchymal transition(EMT)in human breast cancer MDA-MB-231 cells.Methods:XJW-containing serum was prepared by intragastric administration of XJW(4 g·kg-1)to SD rats.MDA-MB-231 cells were cultured in vitro.β-Catenin overexpression and knockdown plasmids were constructed and transfected into MDA-MB-231 cells.The optimal XJW-containing serum(4 g·kg-1)according to previous studies was used for β-catenin intervention and overexpression experiments.The groups for intervention/overexpression experiments included intervention/overexpression blank group,β-catenin intervention/overexpression group,intervention/overexpression blank+XJW group,and β-catenin intervention/overexpression+XJW group.Cell morphology was observed under a microscope.Flow cytometry was used to detect and record apoptosis rates.Western blot was performed to measure apoptosis-related proteins[B-cell lymphoma 2(Bcl-2),Bcl-2-associated X protein(Bax),and active-Caspase-3]and EMT-related proteins(E-cadherin,N-cadherin,and vimentin).Real-time polymerase chain reaction(Real-time PCR)was used to detect β-catenin mRNA expression levels in different groups.Results:Intervention experiment results showed that compared with the intervention blank group,the β-catenin intervention group and the intervention blank+XJW group showed increased apoptosis rates(P<0.05,P<0.01),elevated protein levels of Bax,active Caspase-3,and E-cadherin(P<0.05,P<0.01),and decreased protein levels of Bcl-2 and Vimentin(P<0.05,P<0.01).β-catenin mRNA expression was reduced in the β-catenin intervention group(P<0.05,0.63±0.15)and the intervention blank+XJW group(P<0.05,0.79±0.33).Compared with the β-catenin intervention group,the β-catenin intervention+XJW group exhibited an increased apoptosis rate(P<0.01),elevated Bax and active Caspase-3 protein levels(P<0.05,P<0.01),and decreased Bcl-2,N-cadherin,and Vimentin protein levels(P<0.05).β-catenin mRNA expression was further reduced in the β-catenin intervention+XJW group(P<0.01,0.22±0.08).Compared with the intervention blank+XJW group,the β-catenin intervention+XJW group showed an increased apoptosis rate(P<0.01),elevated Bax,active Caspase-3,and E-cadherin protein levels(P<0.05,P<0.01),and decreased Bcl-2,N-cadherin,and vimentin protein levels(P<0.05,P<0.01).β-catenin mRNA expression was also reduced(P<0.05).Overexpression experiment results showed that compared with the overexpression blank group,the β-catenin overexpression group exhibited a reduced apoptosis rate(P<0.01),while the overexpression blank+XJW group showed an increased apoptosis rate(P<0.05).In the β-catenin overexpression and β-catenin overexpression+XJW groups,Bax and E-cadherin protein levels decreased(P<0.05,P<0.01),while Bcl-2,N-cadherin,and vimentin levels increased(P<0.05).In the overexpression blank+XJW group,Bax and E-cadherin levels increased(P<0.05,P<0.01),while Bcl-2,N-cadherin,and vimentin levels decreased(P<0.05,P<0.01).β-catenin mRNA expression increased in the β-catenin overexpression group(P<0.01,7.26±1.03)but decreased in the overexpression blank+XJW group(P<0.05,0.73±0.14).Compared with the β-catenin overexpression group,the β-catenin overexpression+XJW group showed an increased apoptosis rate(P<0.01),elevated Bax and E-cadherin levels(P<0.05),and decreased N-cadherin and vimentin levels(P<0.05).β-catenin mRNA expression was reduced(P<0.05,4.65±1.02).Compared with the overexpression blank+XJW group,the β-catenin overexpression+XJW group exhibited a reduced apoptosis rate(P<0.01),decreased Bax and E-cadherin levels(P<0.01),and increased Bcl-2,N-cadherin,and Vimentin levels(P<0.05).β-catenin mRNA expression increased(P<0.01).Conclusion:XJW-containing serum inhibits breast cancer cell proliferation by suppressing EMT,which is associated with the regulation of the Wnt/β-catenin pathway.