The Proliferation Inhibitory Effects Of Icotinib And MK-2206 On Hep-2 Cells Of Laryngeal Carcinoma

ObjectiveTo study the proliferation inhibitory effect of Icotinib and MK-2206 on Hep-2cells of human laryngeal squamous cell carcinoma in vitro.To observe influence of the cell cycle distribution and apoptosis of two drugs on Hep-2 cell lines.Provide the experimental basis for application of two drugs in laryngeal cancer treatment.MethodsCulture of human laryngeal squamous carcinoma hep-2 cells In vitro,divided into four groups:the control group,Icotinib group,the MK-2206 group and the combination group.the MTT method was used to detect the proliferation of cells in each group on 24,48,72 hours,and calculate the cell proliferation inhibition rate.48 hours after administration,cell apoptosis and cell cycle status of 4 groups were detected by flow cytometry.The expression levels of PI₃K,Akt and p-Akt protein in each group were detected by Western Bolt,on 48 hours after administration.Results were analyzed with spss19.0 software for one-way anova and S-N-K,P<0.05 indicating that the results are different,the significance threshold was set at P<0.01.ResultsMTT results showed that Icotinib and MK-2206 could inhibit the proliferation of Hep-2 cells,combination group was better than both of Icotinib group and MK-2206 group,and there were two relationship have been found,one of them was a time and dose dependent relationship in two drugs,another was they have synergistic effect.FCM results showed that Icotinib and MK-2206 could change Hep-2 cell cycle distribution,rise G₀/G₁ phase cell ratio,S phase cell ratio is reduced,have significant blocking effect at G₀/G₁ phase,compared with the control group both of them cloud inducing Hep-2 cells apoptosis,and combination was better than alone.Western blot showed that compared with the control group,Icotinib and MK-2206 could significantly reduce the expression of PI₃K,Akt andp-Akt protein,and the combination effect was more obvious.ConclusionsIcotinib and MK-2206 have proliferative inhibitory effects on Hep-2 cell lines and there is a synergistic effect between them.Icotinib and MK-2206 can change cell cycle distribution of Hep-2 cell lines of laryngeal carcinoma,block cell cycle and accelerate apoptosis,which cause the inhibition of proliferation.