Abstract： Objective The aim of this study is to investigate the effects and molecular mechanisms of Juanliu Sanjie Formula(蠲瘤散结方)on the proliferation and glycolysis of lung adenocarcinoma A549 cells.Methods The cell counting kit-8(CCK-8)method was used to detect the cell viability of A549 cells treated with different concentrations of Juanliu Sanjie Formula(2,4,6,8,10,12 mg·mL-)for 24 h and 48 h.A549 cells were divided into control group and Juanliu Sanjie Formula low and high-concentration groups(3,6 mg·mL-1).Drug intervention was added for 24 hours and the proliferation of A549 cells was detected through cell clone formation experiments.A549 cell cycle was detected by flow cytometry.The reagent kit detected glucose con-sumption and lactate production levels in the cell supernatant.The potential of hydrogen(pH)value of cell supernatant was detec-ted by using a pH meter.CCK-8 detected A549 cell proliferation.Real-time RT-PCR(RT-qPCR)and Western blot(WB)were used to detect the expressions of protein kinase B(AKT),phosphorylation protein kinase B(p-AKT),hypoxia inducible factor-1α(HIF-1α),facilitative glucose transporter(Glut1),hexokinase 2(HK2),pyruvate kinase isozyme type M2(PKM2),lactate dehydrogenase A(LDHA)genes and proteins.Results Juanliu Sanjie Formula inhibited the proliferation of A549 cells(P＜0.05,P＜0.01).Compared with the control group,the Juanliu Sanjie Formula low and high concentrations groups(3,6 mg·mL-1)lowered the cell clone formation rate(P＜0.05,P＜0.01),the G0/G1 phase cell ratio(P＜0.05,P＜0.01),the glucose consumption and lactic acid production in the cell supernatant(P＜0.05,P＜0.01).The expressions of AKT(p-AKT),Hif-1α,Glut1,HK2,PKM2,LDHA genes and proteins decreased(P＜0.05,P＜0.01).After using AKT activator,compared with the Juanliu Sanjie Formula high concentration group,the Juanliu Sanjie Formula combined with AKT activator increased glu-cose consumption and lactic acid production in cell supernatant(P＜0.05,P＜0.01),decreased pH value(P＜0.05)and in-creased cell proliferation activity(P＜0.05).The expressions of AKT/p-AKT,Hif-1α,Glut1,HK2,PKM2,LDHA genes and proteins increased(P＜0.05).Conclusion Juanliu Sanjie Formula has inhibitory effects on the proliferation and glycolysis of A549 cells,which may be related to the regulation of AKT signaling pathway.