Abstract: Objective：To analyze the effect of high intensity focused ultrasound (HIFU) combined with sequential radiotherapy on pancreatic cancer via transplanted tumor mice model. Methods：PANC-1 cells were injected subcutaneously into the right scapular region of twenty-four BALB/c nu-nu mice. The mice were then randomly divided to 4 groups, control group, HIFU alone group, RT alone group, HIFU combined RT group. The tumor volume change and acute side reaction were measured. The expressions of Ki-67, Caspase-3, STAT-3, and HMGB-1 were analyzed using immunohistochemistry. Results：In the control group, tumor volume increased significantly. Tumor control rate was 36.32% in HIFU alone group, 84.49% in RT alone group, and 70.49% in HIFU combined RT group, respectively (P＜0.05). There was no acute adverse event in control and RT alone groups. Two shock reaction and four cases of burn injury reaction were all found in HIFU and HIFU combined RT groups. The expression of HMGB-1 was significantly increased in HIFU and HIFU combined RT groups as compared with control and RT alone groups (P＜0.05). Ki-67 and STAT-3 were highly expressed in all groups, but the expression of Caspase-3 was quite low in all groups. Conclusions：The treatment of HIFU combined with sequential radiotherapy on pancreatic cancer failed to show synergistic effect, and the adverse reaction was higher than that of radiotherapy alone.