Alumina nanoparticles (Al
2
O
3
NPs) are gradually used in various areas, including nanomedicine, biosensors, and electronics. The current study aimed to explore the DNA damage and cytotoxicity due to Al
2
O
3
NPs on human hepatocarcinoma cells (HepG2). The MTT and neutral red uptake assays showed that Al
2
O
3
NPs induce significant cell death in a dose- and time-dependent manner. However, Al
2
O
3
NPs induced significant intracellular reactive oxygen species production and elevated lipid peroxidation and superoxide dismutase levels in the HepG2 cells. Al
2
O
3
NPs also induced significant decrease in reduced glutathione levels and increase caspase-3 activity in HepG2 cells. DNA fragmentation analysis using the alkaline single-cell gel electrophoresis showed that Al
2
O
3
NPs cause genotoxicity in dose- and time-dependent manner. However, they induce reactive oxygen species production and oxidative stress, leading to oxidative DNA damage, a probable mechanism of genotoxicity. This study warrants more careful assessment of Al
2
O
3
NPs before their industrial application.