This study was conducted to investigate the physicochemical, phytochemical, in vitro antidiabetic and anticancer potential of Olea ferruginea R bark. After extraction using Soxhlet, in vitro antidiabetic and cytotoxic activity on human hepatocellular carcinoma (HepG2) cells was assessed by nonenzymatic glycosylation of hemoglobin assay, alpha-amylase inhibition assay, glucose uptake by yeast cells, and 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide assay, respectively, and gene expression via real-time polymerase chain reaction. Primary and secondary metabolites were present in the extractants; polyphenols (35.61 +/- 0.03) and flavonoids (64.33 +/- 0.35) in the chloroform; and polysaccharides in the ethanol (268.75 +/- 0.34), and glycosaponins (78.01 +/- 0.07) in the methanol. The chloroform extract exhibited maximum antidiabetic potential, showing inhibition of nonenzymatic glycosylation of hemoglobin (65%), and alpha-amylase inhibition (32%) with maximum percent glucose uptake by the ethanol extract (78%). Only the ethanol extract had dose-dependent cytotoxic potential against the HepG2 cells. After 24-h exposure to the ethanol-extract, the expression of protein kinase B (Akt) remained unchanged, while the expression of B-cell lymphoma 2 (BCL2) and BCL2 associated X (BAX) changed significantly. After 48-h exposure, the expression of Akt decreased significantly, while that of BCL2 and BAX increased significantly. Olea ferruginea R bark possessed in vitro antidiabetic potential and anticancer/cytotoxic effects, attributable to the decline in the prosurvival signals of the Akt signaling pathway.