Polydopamine nanoparticles (PDA NPs), among nature-inspired building materials, show special functions for biomedical systems and exploring PDA derived nanostructures for future developments is a fast growing field. Herein, we demonstrated the first evaluation of the PDA NPs for the electrochemical determination of lung cancer cells. In the presented study, PDA NPs were synthesized in a mild and cost-effective fashion by self-polymerization of dopamine in an alkaline environment. The structural and chemical characterizations clearly demonstrated the formation of PDA NPs with controllable size (130 nm), hence applied as a suitable material to functionalize the pencil graphite electrode (PGE) surface to construct a cytosensing nanoprobe. The ability of the developed sensor (PDA NPs/PGE) for label-free electrochemical A-549 lung cancer cells detection was investigated. The designed PDA NPs based cytosensor exhibited good biocompatibility and sensitivity for impedimetric diagnosis of A-549 cells in a wide linear range (1.0 x 10(2)-1.0 x 10(5) cells mL(-1)) with low detection limit (25 cells mL(-1)). Furthermore, the developed bioassay has great potential as liquid biopsy for early cancer detection.