Abstract: Objective To investigate the effect and mechanism of Hedyotis diffusa-sculellaria barbata components on the proliferation, invasion, migration and apoptosis of adenocarcinoma of colon Lovo cells. Methods The Hedyotis diffusa and scutellaria barbata were decocted for three times according to the quality of 1∶1 to obtain the water extract. Then, a proper amount of extract was taken and degreased with petroleum ether, and then extracted with ethyl acetate for many times to obtain the components of Hedyotis diffusa and scutellaria barbata and calculated the yield. The experimental cells were divided into control group (normal cultured Lovo cells), Hedyotis diffusa-sculellaria barbata component low dose group (10 mg/L), medium dose group (30 mg/L), and high dose group (50 mg/L). The proliferation inhibition rates were detected by thiazole blue colorimetric (MTT) assay. The invasion ability and migration ability were checked by Transwell and cell scratch test. Flow cytometry was used to detect cell cycle distribution, and TUNEL was used to check apoptosis rate. The expression of Grb2-associated binding protein 1 (Gab1)/Vascular endothelial growth factor receptor 2 (VEGFR-2)/Phosphatidylinositol 3-kinase (PI3K)/Kthreonine kinase (Akt) signaling pathway protein, apoptotic protein Bcl-2 and Bax and metalloproteinase-9 (MMP-9) were detected by Western blot assay. Results Four compounds, p-hydroxyacetophenone, scutellarin, luteolin and apigenin, were found in the pair of Hedyotis diffusa and Scutellaria barbata. The yield was 0.61% after ethyl acetate extraction. Compared with the control group, the cell proliferation inhibition rate, apoptosis index (AI) value, the proportion of G1 phase of tumor cells increased in the low, middle and high dose groups, the number of cell penetrating membrane and the rate of scratch closure decreased significantly (P＜0.05). The expressions of Gab1, VEGFR-2, PI3K, Akt, MMP-9 and Bcl-2 proteins decreased, the expression of Bax increased with a dose-dependent manner (P＜0.05). Conclusion Hedyotis diffusa-sculellaria barbata components can inhibit the proliferation of adenocarcinoma of colon Lovo cells in G1 phase, reduce the migration and invasion and induce the apoptosis. The mechanism might be related to the inhibition of Gab1/VEGFR-2/PI3K/Akt signaling pathway.