Abstract: Objective To investigate the effect of Icotinib hydrochloride on PI3K/AKT and HIF-1α signaling pathway in lung adenocarcinoma A549 cell. Methods The lung adenocarcinoma A549 cells were cultured and treated with different concentration of Icotinib hydrochloride.The cell counting was used to assess the cell growth,MTT to evaluate the cell proliferation,microscope to observe the changes of cellular morphology,TUNEL to test the apoptosis rate,and immunohistochemistry to determine the expression of PI3K/AKT and HIF-1α protein. Results The growth of tumor cells inhibited significantly with irregular configuration when the concentration of Icotinib hydrochloride increased.And at 50 mg/L,a large number of GNM cells turned into necrosis.Icotinib hydrochloride inhibited the proliferation in a dose- and time-dependent manner.The treatment groups were different from the control group with significant difference(P＜0.01);The apoptosis rate in 12.5 mg/L group,25 mg/L group,50 mg/L group,DPP(25 mg/L) group were significant higher than that in the control group,the differences were statistically significant(P＜0.05);the expression of PI3K/AKT and HIF-1α protein of A549 in 5 mg/L group,50 mg/L group,DPP(25 mg/L) group were significantly lower than that in the control group,the differences were statistically significant(P＜0.05). Conclusion Icotinib hydrochloride has inhibitory effect on the proliferation of human lung adenocarcinoma A549 cell,the mechanism of action of Icotinib hydrochloride may involve inducing apoptosis and inhibiting of PI3K/AKT and HIF-1α signaling pathway.