The Intervention Effect Of EEHD And Gefitinib On The Different Phenotypes Lung Adenocarcinoma Cells With Epithelial-mesenchymal Transition In Vitro

Objective:To study the effect of EEHD combined with Gefitinib on the proliferation of different phenotypes lung Adenocarcinoma cells(A549、HCC827)and expression of E-cadherin、Vimentin、EGFR and cell apoptosis in the EMT cells induced by TGF-(31.Methods:Detected the proliferative effect of EEHD and Gefitinib on A549、HCC827 cells with CCK8.Induced EMT in A549、HCC827 cells with TGF-β1 and observed the cell morphology under the inverted microscope.Detected the protein expression of epithelial marker(E-cadherin)and mesenchymal cell marker(Vimentin)with Western Blot.The cells were divided into some groups and added with EEHD、Gefitinib、EEHD combined with Gefitinib in different orders.Assay the the protein expression of E-cadherin、Vimentin、EGFR by Western Blot and detected apoptosis with flow cytometry technology.Results:1.CCK8 assay showed that EEHD and Gefitinib inhibited the proliferation of A549 and HCC827 in a concentration dependent manner.The IC50 value of EEHD treating A549 and HCC827 cells was 404.51,887.85μg/ml.The IC50 of Gefitinib treating A549 and HCC827 cells was 24.046,27.832μg/ml.2.TGF-β1 could lead the cells to induce EMT..2.The date showed,to A549 cells,the experimental groups had higher expression of E-cadherin than control group(P<0.05)and EEHD combined with Gefitinib group showed lower expression of Vimentin than control group and Gefitinib group(P<0.05).To the HCC827 cells,the EEHD combined with Gefitinib group showed lower expression of Vimentin than control group(P<0.05)and EEHD combined with Gefitinib 48h group showed lower expression than Gefitinib group(P<0.05).Then the experimental groups showed higher expression than control group(P<0.05)and the EEHD combined with Gefitinib groups showed higher expression than Gefitinib group(P<0.05).3.According to result of the flow cytometry,the apoptosis rate of experimental groups was higher than that of control group(P<0.05).The rate of apoptosis of EEHD combined with Gefitinib group was higher than that of the other experimental groups(P<0.05)and the rate of apoptosis of EEHD combined with Gefitinib in different orders had no statistical significance(P>0.05).Conclusion:1.The anti-proliferative effect of EEHD and Gefitinib on A549 and HCC827 were different,and the anti-proliferative effect on epithelial phenotypic cell A549 was stronger than that of HCC827 which was interstitial phenotype.2.The EMT model of HCC827 and A549 cells could be induced by TGF-β1.3.The protein expression of epithelial marker(E-cadherin)in EMT cells could be up-regulated and mesenchymal cell marker(Vimentin)could be down-regulated by the intervention of EEHD combined with Gefitinib in different orders.4.The apoptosis on EMT cells could be increased by the treatment of EEHD combined with Gefitinib in different orders.And EEHD combined with Gefitinib had the strongest ability to increase the apoptosis.