Abstract：According to the 2018 Global Cancer Statistics Survey, colon cancer account for 12.1% of new cancer cases. Therefore, it is urgent to find new genes related to the occurrence of colon cancer and new therapeutic targets. By analyzing the database, we found that the expression levels of RTN4 was significantly correlated with the survival rate of colon cancer patients. In this study, the interfering plasmid of RTN4 gene was constructed, and lentivirus was used as the vector to transfect colon cancer HCT116 cells to construct colon cancer cell lines with RTN4 knock-down. The results showed that RTN4 knock-down significantly promoted the proliferation of colon cancer cell HCT116. Western blot was used to observe the expression of autophagy pathway related proteins p62 and LC3 in HCT116 cells after RTN4 knock-down. It was found that the LC3 transformation amount (LC3-II/LC3-I) was increased and the p62 protein was decreased in the RTN4 knockdown group compared with the control group. In this study, the potential anticancer effects of RTN4 was analyzed. Knockdown of RTN4 gene significantly enhanced the proliferation of colon cancer cells, promoted the proliferation of colon cancer cells, and induced autophagy, which may be related to the activation of LC3/p62 autophagy pathway.