枸橘-壁虎通过调控PI3K/Akt通路对三阴性乳腺癌细胞增殖、凋亡和上皮间质转化的影响OA

Objective It is to investigate the effects and mechanism of Poncirus trifoliata with Gecko on the prolifera-tion,apoptosis and epithelial-mesenchymal transition(EMT)of triple negative breast cancer(TNBC)4T1cells.Methods 4T1 cells were taken and cultured,and their cell viabilities after treatment with different concentrations of Poncirus trifoliata with Gecko were assesed by CCK-8 assay to determine the appropriate concentration for subsequent experiments.A blank group and low-dose,medium-dose,and high-dose groups of Poncirus trifoliata with Gecko were set up in the experiment.The cell migration ability was assessed by scratch assay(cell scratch healing rate was calculated),the cell apoptosis rate was determined by flow cytometry,the mRNA expressions of EMT-related markers in cells,including N-cadherin,vimentin and E-cadherin,as well as phosphoinositide 3-kinase(PI3K)and protein kinase B(Akt)were detected by and qPCR,the pro-tein expressions of N-cadherin,vimentin,E-cadherin,as well as PI3K,p-Akt and Akt(these three indicators were addition-ally detected in the high-dose Poncirus trifoliata with Gecko+SC79 group)were detected by Western blot analysis.Results After 24 hours of culture,the IC50 value of Poncirus trifoliata with Gecko in the treatment of 4T1 cells was 8.069 mg/mL,4,6,and 8 mg/mL were selected as the low,medium,and high doses of Poncirus trifoliata with Gecko for the subsequent studies.Compared with the blank group,the cell scratch healing rate and the relative mRNA expressions of N-cadherin,vi-mentin,and Akt,as well as the relative protein expressions of N-cadherin,vimentin,PI3K,p-Akt and Akt in each dose groups of Poncirus trifoliata with Gecko were significantly reduced(all P<0.05),while the cell apoptosis rate,relative mRNA expression of E-cadherin in the cells of each does groups of Poncirus trifoliata with Gecko,and the relative protein expressions of E-cadherin in the cells of the medium-dose and high-dose groups of Poncirus trifoliata with Gecko were signif-icantly increased(all P<0.05),and the improvements of all indicators(except for the relative mRNA expression of PI3K)in the high-dose group of Poncirus trifoliata with Gecko were more significant than those in the low-dose and medi-um-dose groups(all P<0.05).The relative protein expressions of PI3K,p-Akt and Akt in the high-dose Poncirus trifolia-ta with Gecko+SC79 group were all significantly higher than those in the high-dose group of Poncirus trifoliata with Gecko(all P<0.05).Conclusion Poncirus trifoliata with Gecko can inhibit the proliferation,migration and EMT of 4T1 cells,and promote their apoptosis,the mechanism may be related to the inhibition of PI3K-Akt pathway.