Objective To investigate the apoptosis, immunity response and angiogenesis factor changes in rabbits with pancreatic cancer xenografts after treatment with irreversible electroporation (IRE,commonly known as nanoknife), cryoablation, or radiofrequency ablation (RFA), and to observe the antitumor effects of the above minimally invasive techniques for pancreatic cancer xenografts. Methods Eight pancreatic cancer xenograft model rabbits were randomly divided into the control, IRE, cryoablation, and RFA groups, with each group containing 2 rabbits. The vital signs of the animals were observed during the treatment and all the rabbits were executed on the next day after treatment. The serum samples were collected to examine the liver, kidney function and creatine kinase levels; and ELISA method was used to detect the cardiac troponin I(CTnI), Caspase-3, tumor necrosis factor-α (TNF-α) and vascular endothelial growth factor(VEGF) levels in the plasma. Morphological changes of the tumor tissues were subjected to H-E staining, and Tunnel assay was used to detect the apoptosis of tumor cells. Immunohistochemistry method was applied to detect Bcl-2, HSP70 and VEGF expression in pancreatic cancer xenograft tissues. Results All the rabbits survived after treatment. The tumors had a clear border and were hard in texture. Obvious coagulative necrosis areas were seen in the treatment areas in each treatment group. H-E staining showed a clear borderline between the necrotic and normal areas, which was especially true in the IRE group. A large number of red blood cells and inflammatory cell infiltration were seen around the border of the necrosis in each group, with some live cells seen in the cryoablation and RFA groups. Tunnel assay showed that RFA significantly increased the apoptosis in pancreatic cancer cells compared with the other two treatment groups. The plasma Caspase-3,TNF-α and VEGF levels were increased in each treatment group. Bcl-2 and VEGF expression was decreased and HSP70 expression was increased in the treatment areas. Post-treatment serological test showed that serum amylase, liver, kidney functions and CTnI were all within the normal range, except for a significantly higher level of creatine kinase in the IRE group. Conclusion IRE, cryoablation, and RFA can inhibit pancreatic cancer xenograft through inducing apoptosis, producing specific antitumor immune response and inhibiting angiogenesis; they are safe and effective. IRE has certain advantages in producing specific antitumor immune response and protecting important peripheral organs.