Synergic Effects of Carthamin and miR-214 on Proliferation and Apoptosis of Lung Cancer Cells
Objective:To investigate the effects of carthamin combined with miR-214 on the prolifera-tion and apoptosis of lung cancer cells.Methods:Quantitative real-time PCR was conducted to determine miR-214 expression in human normal lung epithelial cells BEAS-2B and lung cancer cells A549 and H1299.miR-214 mimic(miR-214)and negative control(miR-NC)were transfected into A549 and H1299 cells.A549 and H1299 cells were divided into control,carthamin,miR-214 and carthamin+miR-214 groups.Cells in carthamin group were treated with 80μmoL/L carthamin,cells in miR-214 group were transfected with miR-214,and cells in carthamin+miR-214 group were co-treated with carthamin and miR-214.The cells without any treatment were used as control group.CCK-8,EdU and flow cytometry assays were per-formed to evaluate cell viability,proliferation,cell cycle and apoptosis,respectively.The protein expressions were analyzed with Western blot.A xenograft tumor model was used to evaluate the synergy of carthamin and miR-214 on lung cancer growth in vivo.Results:Compared with BEAS-2B cells,miR-214 expression was significantly reduced in A549 and H1299 cells.miR-214-transfected A549 and H1299 cells expressed higher level of miR-214,compared to miR-NC.Compared with control group,cell viability,EdU-positive cells and S-phase cells were markedly reduced,while G0/G1-phase cells and apoptotic cells were substantially in-creased in carthamin and miR-214 groups;the cell viability,EdU-positive cells and S-phase cells were less,while the G0/G1-phase cells and apoptotic cells were more in carthamin+miR-214 group than those in carthamin and miR-214 groups.Compared with control group,the phosphorylation of Akt and PI3K and pro-tein expressions of Bcl-2,Cyclin D1 and CDK4 were considerably downregulated,while the expressions of Bax and cl-Caspase-3 were significantly up-regulated in carthamin and miR-214 groups.Akt and PI3K phosphorylations and protein levels of Bcl-2,Cyclin D1 and CDK4 in carthamin+miR-214 group were much lower,while Bax and cl-Caspase-3 expressions were higher than those in carthamin and miR-214 groups.In vivo experiments showed that compared with control group,the tumor volume and weight of mice were signifi-cantly reduced in carthamin and miR-214 groups,which were further smaller in carthamin+miR-214 group than those in carthamin and miR-214 groups.Conclusion:Carthamin combined with miR-214 could inhibit the proliferation but promote apoptosis of lung cancer cells in vitro and in vivo,probably through the inactiva-tion of PI3K/Akt signaling pathway.
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