Objective To investigate the anti-canter effects of inert gas microbubbles combined with low frequency and low intensity ultrasound irradiation on human pancreatic cancer PANC-1 cells. Methods The human pancreatic cancer PANC-1 single cell suspension with concentration 1 × 106 /ml were divided into 4 groups. The the blank control group was added with 2 ml human pancreatic cancer PANC-1 single cell suspension. The inert gas microbubble group was added 1600 μL human pancreatic cancer PANC-1 single cell suspension and 400 μL inert gas microbubbles and fully mixed to make the concentration of inert gas microbubbles 20%. In the low frequency and low intensity ultrasound group, 2 ml of human pancreatic cancer PANC-1 single cell suspension was added, and then irradiated with low frequency and low intensity ultrasound continuous wave with sound intensity of 0.60 w/cm2 and frequency of 1 MHz for 60 s. The microbubbles + ultrasound group was added 1600 μL human pancreatic cancer PANC-1 single cell suspension and 400 μL of inert gas microbubbles and fully mixed so that the concentration of inert gas microbubbles was 20%, and then irradiated continuously with low frequency and low intensity ultrasonic continuous wave with sound intensity of 0.60 w/cm2 and frequency of 1 MHz for 60 s. The proliferation activity of cells in each group was detected by cell counting reagent method, and the apoptosis of cells in each group was measured by analytical flow cytometry. Results The proliferative activity of human pancreatic cancer PANC-1 cells in the blank control group,the inert gas microbubble group, the low frequency low intensity ultrasound group and thw microbubble + ultrasound group were 0.9356 ± 0.1652, 1.1126 ± 0.0738, 0.3236 ± 0.0126 and 0.1566 ± 0.0137 respectively. The proliferative activity of the microbubble + ultrasound group was lower than that of the other groups, and the difference was statistically significant (all P < 0.05). The apoptosis rates of the blank control group, the inert gas microbubble group, the low-frequency low-intensity ultrasound group and the microbubble + ultrasound group were 0.069 ± 0.007, 0.033 ± 0.006, 0.279 ± 0.016 and 0.678 ± 0.018 respectively. The apoptosis rate of microbubble + ultrasound group was higher than that of the other groups, and the difference was statistically significant (all P < 0.05).Conclusion The inert gas microbubbles combined with low frequency and low intensity ultrasound irradiation can reduce the proliferation of human pancreatic cancer PANC-1 cells cultured in vitro, induce apoptosis of cancer cells, and have obvious anti-canter effects, providing a new research idea for tumor therapy.