Baicalein Suppresses HEGF-induced Proliferation And Migration Of Glioma Cells Via The EGFR/Akt Signaling Pathway

Objective: 1.To observe the inhibitory effect of baicalein(BAI)on the proliferation and migration of human glioma cell line U251 and its effect on the phosphorylation of protein EGFR and Akt.2.To explore the mechanism of baicalein on human glioma cell line U251 and its clinical significance.Methods: 1.Human glioma cell lines U251 were treated with human epidermal growth factor(hEGF,20 ng/ml)or epidermal growth factor(EGFR)inhibitor—AG1478(10 ?M)or combination at different time(12 h,24 h,36 h and 48 h)for measuring cell proliferation by CCK assay;2.Human glioma cell lines U251 were treated with hEGF(20 ng/ml)or EGFR inhibitor—AG1478(10 ?M)or combination at 24 h for measuring cell migration by Wound-healing assay;3.Human glioma cell lines U251 were treated with hEGF(20 ng/ml)or EGFR inhibitor—AG1478(10 ?M)or combination at 24 h for detecting the phosphorylation of EGFR and Akt proteins by western blotting assay;4.Human glioma cell lines U251 were treated with BAI(20 ?M,40 ?M and 80 ?M)or hEGF(20 ng/ml)or combination at different time(12 h,24 h,36 h and 48 h)for measuring cell proliferation by CCK assay;5.Human glioma cell lines U251 were treated with BAI(20 ?M,40 ?M and 80 ?M)or hEGF(20 ng/ml)or combination at 24 h for measuring cell migration by Wound-healing assay;6.Human glioma cell lines U251 were treated with BAI(10 ?M,20 ?M,40 ?M and 80 ?M)or hEGF(20ng/ml)or combination at different time(6 h,12 h,24 h and 36 h)for detecting the phosphorylation of EGFR and Akt proteins by western blotting assay;7.Human glioma cell lines U251 were treated with BAI(80 ?M)or hEGF(20 ng/ml)or combination at 24 h for detecting the phosphorylation of EGFR and Akt proteins by Immunofluorescence staining.Results: 1.the phosphorylation of EGFR and Akt proteins in U251 cells were promoted by treated with hEGF at 24 h;2.The proliferation of U251 cells were promoted by treated with hEGF at different time(12 h,24 h,36 h and 48 h)in a time-dependent manner;3.the migration of U251 cells were promoted by treated with hEGF at 24 h;4.the phosphorylation of EGFR and Akt proteins in U251 cells were inhibited by treated with different concentrations BAI(10 ?M,20 ?M,40 ?M and 80 ?M)at 24 h,and BAI prevent the promotion of hEGF in a dose-dependent manner;5.the phosphorylation of EGFR and Akt proteins in U251 cells were inhibited by treated with BAI(80 ?M)at different time(6 h,12 h,24 h and 36 h),and BAI prevent the promotion of hEGF in a time-dependent manner;6.the proliferation of U251 cells were inhibited by treated with different concentrations BAI(20 ?M,40 ?M and 80 ?M)at different time(12 h,24 h,36 h and 48 h),and BAI prevent the promotion of hEGF in a time-and dose-dependent manner;7.the migration of U251 cells were inhibited by treated with different concentrations BAI(20 ?M,40 ?M and 80 ?M)at 24 h,and BAI prevent the promotion of hEGF in a time-dependent manner;Conlusion:1.hEGF promotes the proliferation and migration of U251 cells throughEGFR / Akt signaling pathway;2.BAI suppresses hEGF-induced proliferation and migration of U251 cells via the EGFR/Akt signaling pathway.