The Podocarpus Nagi kernel oil was prepared and its chemical constituents were confirmed accordingly in this work. The constituents and the contents of the obtained oil were identified as: total flavonoids (4.2%, calculated according to the rutin), (9Z,12Z)-9,12-octadecadienoic acid(38.2%), arachidic acid(0.11%), cis-11-eicosenoic acid(1.40%), cis-11,14-eicosadienoic acid(8.04%), ꞷ-3(0.2%), ꞷ-6(48.0%), ꞷ-9(21.0%), vitamin E(2.11mg/100g). The microelements in this oil was also tested using ICP-MS and the results exhibited that this oil contains numbers of microelements beneficial for human health: such as V(0.006μg/g), Cr(0.024μg/g), Mn(40.109μg/g), Fe(2.292μg/g), Co(0.007μg/g), Zn(4.316μg/g), As(0.009μg/g), Se(0.240μg/g), Sr(0.453μg/g). Then, the oil was evaluated for its in vitro antioxidant effect against DPPH⸱, which exhibited noteworthy scavenging ability against DPPH⸱. In addition, the oil was also evaluated for its preliminary in vitro anticancer activity against four cancer cell lines: the results showed that it exhibited the highest inhibition against gastric cancer, breast cancer (MCF-7), lung cancer(A549) and Hela cell lines with the highest inhibitions of 64.30%±2.80, 52.87%±2.57, 93.21%±2.24 and 88.33%±2.08 at the concentrations of 25mg/mL, 25mg/mL, 50mg/mL and 50mg/mL, and with the IC50s of 48.47mg/mL, 763.14mg/mL, 11.47mg/mL and 23.77mg/mL respectively. These findings demonstrated that this oil can be regarded as the functional edible oil by comparing the contents of edible oils used in the field of food.