Abstract: Objective To establish the cell model of LOVO with overexpressed AT motif binding factor 1 (ATBF1), and to observe the change of the proliferation, apoptosis, invasion of LOVO cell line. Methods The result was detected under a fluorescent microscope after pEGFP-ATBF1 was transfected into human colorectal carcinoma cell line LOVO by Lipofectamine. The expression of ATBF1 and Notch3 were identified by RT-PCR and Western blot. CCK-8 method was used to measure cell growth and Transwell migration assay was utilized to detect the invasion of overexpressed ATBF1 LOVO. The relationship between ATBF1 and the apoptosis of LOVO cells was analyzed by flow cytometry. Results Compared with blank group and control group, the proliferation and invasion of overexprssed ATBF1 LOVO were inhibited in the trial group (P<0.001). There were higher apoptosis rate of LOVO cells (15.9%) in treatment group than those in blank(5.2%) and control groups(4.6%) according to the result of flow cytometry. However, Western blot showed that the expression of oncogene Notch3 descended in the trial group (P<0.05). Conclusion Anti-oncogene ATBF1 could inhibit the proliferation and invasion of LOVO cells and induce its apoptosis, meanwhile, ATBF1 could downregulate the expression of Notch3, which means Notch3 might be one of the downstream gene of ATBF1.