The aim of this research was to extract and characterize the volatile oil from Artemisia abyssinica using the hydro-distillation method, then use this to evaluate antimicrobial activity by means of molecular docking. The results of the GC/MS analysis showed that the sample contained 25 different compounds, belonging to various classes, such as: oxygenated sesquiterpene (14.75%), aromatics (7.6%), esters (6.9%) as well as one ketone (5.21%) and butyl isothiocyanate (4.38%). The most abundant compounds were: γ – terpinene (20.02%), camphor (15.75%), camphene (9.86%), nerolidol (9.54%), -3-carene (8.85%), davanone (5.21%) and isobutyl isothiocyanate (4.38%). The oil exhibited antimicrobial properties that protected against some of the strains tested (E. coli, Kl. Pneumonia, St. aureus and Str. mutans). The oil inhibited the growth of these microorganisms. The minimum inhibitory concentration (MIC) was determined that protected against four bacterial strains and it exhibited strong antimicrobial activity against Kl. Pneumonia (MIC= 13.3 mg/ml). The results of the antitumor activity of the oil with regard to protecting against hepato cell carcinoma (Hepg2) proved that the oil display a promising antitumor activity with IC50=9.5g/ml. The application of molecular docking substantiated that some of the identified oil components display a significant binding activity against DHFR compared to Gentamycin and Ampicillin antibiotics (positive controls). Furthermore, the molecular docking finding regarding EGFR, which was performed to explain the antitumor activity based on the structural futures, proved that davanone and nerolidol are the major contributors to antitumor activity.