In order to examine bee pollen hydrolysates to assess their anticancer and antioxidant properties, hydrolysis of bee pollen was first performed using three different commercially available enzymes: Alcalase®, Neutrase®, and Flavourzyme®. The study used DPPH and ABTS assay to evaluate the antioxidant properties of the hydrolysates obtained from bee pollen. All of the tested hydrolysates demonstrated antioxidant activity, while hydrolysate based on Alcalase® offered a high value for IC50 and was therefore chosen for further separation into five sub-fractions via ultrafiltration. The greatest antioxidant activity was presented by the MW < 0.65 kDa fraction, which achieved an IC50 value of 0.39 ± 0.01 µg/mL in the DPPH assay and 1.52 ± 0.01 µg/mL for ABTS. Purification of the MW < 0.65 kDa fraction was completed using RP-HPLC, whereupon the three fractions from the original six which had the highest antioxidant activity underwent further examination through ESI-Q-TOF-MS/MS. These particular peptides had between 7 and 11 amino acid residues. In the case of the MW < 0.65 kDa fraction, testing was also carried out to determine the viability of lung cancer cell lines, represented by ChaGo-K1 cells. Analysis of the antiproliferative properties allowed in vitro assessment of the ChaGo-K1 cells' viability following treatment using the MW < 0.65 kDa fraction. Flow-cytometry generated date which revealed that it was possible for the MW < 0.65 kDa fraction to induce apoptosis in the ChaGo-K1 cells in comparison to the results with cells which had not been treated.