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Objective: To observe the radiosensitivity effect of AG825 on breast cancer cell line with high expression of ERBB2 in vitro. Methods: MTT and clone formation assay were used to observe the effect of AG825 on proliferation and radiosensitivity of breast cancer line MDA-MB-453. After MDA-MB-453 was exposed to AG825 and radiation, comet assay and Weste blotting were applied to detect double strand break and expressions DNA-PKcs protein, respectively. Results: AG825 inhibited proliferation rate and decrease survival fraction of MDA-MB-453. After radiation, compared with control group, expression of DNA-PKcs was lower in group with AG825 presence but double strand break was higher. Conclusion: AG825 could increase radiosensitivity of breast cancer cell line MDA-MB-453, and it may associate with its inhibition of radiation induced expression of DNA-PKcs and double strand break repair.