Abstract: Objective To investigate the effect of acidic nuclear phosphoprotein 32A(ANP32A) on the proliferation of colon cancer cells and related mechanisms.Methods The mRNA level of ANP32A in colon cancer and its relationship with patient survival were analyzed by bioinformatics. DLD-1 and HCT-116 cells were used to construct stable lentiviral cell lines with ANP32A silencing, and cell proliferation was detected by CCK-8 and colony formation assays. Myc-ANP32A plasmid was transferred into DLD-1 and HCT-116 cells by transient transfection, and cell proliferation was detected by CCK-8 and colony formation assays. The effect of ANP32A on the expression of high mobility group protein B1 (HMGB1) was detected by qPCR and Western blot. The effect of ANP32A on extracellular regulated protein kinase (ERK) pathway was detected by Western blot. Finally, flag-HMGB1 plasmid was transferred into ANP32A silenced cells, and the levels of the proteins in the ERK pathway and the proliferation ability of colon cancer cells were detected by Western blot and CCK-8.Results The mRNA levels of ANP32A in colon cancer were high and negatively correlated with survival. ANP32A silencing attenuated the proliferation of colon cancer cells, while ANP32A overexpression enhanced the proliferation of the cells. ANP32A enhanced the transcription of HMGB1 and promoted its protein expression. ANP32A promoted the proliferation of colon cancer cells by activating the ERK signaling pathway via HMGB1.Conclusions ANP32A is highly expressed in colon cancer. It can promote HMGB1 protein expression by enhancing its transcription, thereby activates the ERK pathway to promote colon cancer cell proliferation.