〔Abstract〕 Objective To investigate the effects of actin like 6 A(ACTL6 A) knockdown on the proliferation, apoptosis, migration and invasion of SW1990 cells in pancreatic cancer. Methods The Oncomine database was used to analyze the expression of ACTL6 A mRNA in the tissues of pancreatic cancer and normal pancreas. The plasmid of knockdown ACTL6 A and siRNA negative control were established and transfected into SW1990 cell line as siRNA-ACTL6 A group and siRNA-NC group. CCK-8, cell apoptosis experiment, Wound healing and Transwell assay were used to determine the effects of ACTL6 A knockdown on the proliferation, apoptosis, migration and invasion of SW1990 cells. GSEA predicted a possible pathway regulated by ACTL6 A in pancreatic cancer. T-test was used between the two groups. Results The expression of ACTL6 A in pancreatic cancer tissues was higher than that in normal pancreatic tissues(P<0.05). The results of CCK-8 assay showed that the absorbance of siRNA-ACTL6 A group at 24 and 48 h were lower than those in the siRNA-NC group, and the difference was statistically significant(t=5.840, 8.454,P<0.01). The results of Wound healing assay and Transwell assay showed that the healing rate and the number of invasive cells in siRNA-ACTL6 A group were both lower than those in the siRNA-NC group. The difference was statistically significant(t=3.960,4.464,P<0.05), but the apoptosis rate of siRNA-ACTL6 A group was significantly higher than that of the siRNA-NC group, and the difference was statistically significant(t=12.192,P<0.001). GSEA results showed that the group with high expression of ACTL6 A mRNA was up-regulated in cell cycle, nucleotide excision repair, base excision repair, DNA replication, pathways in cancer, NOTCH signaling pathway and other related gene sets(P<0.05). These pathways were activated when the expression of ACTL6 A was up-regulated. Conclusion ACTL6 A is highly expressed in pancreatic cancer tissues. ACTL6 A knockdown promotes the cell apoptosis of SW1990 cells, and inhibits proliferation, invasion and migration of SW1990 cells. The mechanism of the occurrence and development of ACTL6 A in pancreatic cancer is attributed to the activation of cell cycle, nucleotide excision repair, base excision repair, DNA replication, pathways in cancer, NOTCH signaling pathway.