In the present investigation, we detected anti-dsDNA autoantibodies in cancer patients and modeled the production of anti-dsDNA autoantibodies by inoculating tumors in BALB/c mice. Moreover, induction of anti-dsDNA autoantibodies by immunization with inactivated tumor cells and their DNA indicated that DNA of tumor cells was probably the primary antigen, which was supported by the significantly increasing levels of sera free DNA in cancer patients and tumor-bearing mice. cELISA and indirect immunofluorescence assay showed that the anti-dsDNA autoantibodies could bind to the surface components of tumor cells. In vitro assay showed that immunosera at week 6 from immunized mice displayed significant cytotoxicity to tumor cells compared to that of negative control, but no cytotoxicity mediated by immunosera at week 22 was observed. In addition, by flow cytometry and electrophoresis of fragmented DNA, the cytotoxicity might probably be mediated by apoptosis. Our data also showed that the ability of the anti-dsDNA autoantibodies to induce apoptosis of SP2/0 and Wehi 164 cells was significantly correlated (r = 0.990, p < 0.01 and r = 0.901, p < 0.05) with their functional affinity. In vivo, the growth of solid tumors was significantly inhibited in the immunized mice inoculated directly with SP2/0 and Wehi 164 cells, or in the naïve mice which were inoculated with SP2/0 cells preincubated with immunosera containing anti-dsDNA autoantibodies. In conclusion, we demonstrated the origin of anti-dsDNA autoantibodies in cancer patients and tumor-bearing mice. And our data also showed that these autoantibodies revealed anti-tumor effect by inducing apoptosis.