Nanomedicine has been used in tumor treatment and research due to its advantages of targeting, controlled release and high absorption rate. Silver nanoparticle (AgNPs), with the advantages of small particle size, and large specific surface area, are of great potential value in suppressing and killing cancer cells. Methods: AgNPs–polyethyleneimine (PEI) –folate (FA) (AgNPs–PF) were synthesised and characterised by several analytical techniques. The ovarian cancer cell line Skov3 was used as the cell model to detect the tumor treatment activity of AgNPs, AgNPs–PF and AgNPs+ AgNPs–PF. Results: Results shown that AgNPs–PF were successfully constructed with uniform particle size of 50–70 nm. AgNPs, AgNPs–PF, AgNPs–PF+ AgNPs all showed a certain ability to inhibit cancer cell proliferation, increase reactive oxygen species and decrease the mitochondrial membrane potential. All AgNPs, AgNPs–PF, AgNPs+ AgNPs–PF promoted DNA damage in Skov3 cells, accompanied by the generation of histone RAD51 and γ-H2AX site, and eventually leading to the apoptosis of Skov3 cells. The combination of AgNPs–PF and AgNPs had a more pronounced effect than either material alone. Conclusion: This study is to report that the combination of AgNPs+ AgNPs–PF can cause stronger cytotoxicity and induce significantly greater cell death compared to AgNPs or AgNPs–PF alone in Skov3 cells. Therefore, the combined application of drugs could be the best way to cancer treatment.