Objective: To investigate the influence of AFAP-1L2 down-regulation on pancreatic cancer cells and its mechanism. Methods: Human pancreatic cancer SW1990 cells were transfected with AFAP-1L2 targeting plasmids siAFAP-1L2 or negative control siRNA sequences, using untreated SW1990 cells as blank control, and then the migration and invasion ability, and the protein and mRNA levels of the molecules related to metastasis and invasion in each group of cells were detected. The interaction between AFAP-1L2 and p85α protein was tested by co-immunoprecipitation assay. Results: Compared with the blank control SW1990 cells, in SW1990 cells with down-regulated AFAP-1L2 expression after siAFAP-1L2 transfection, the migration and invasion ability was decreased, expressions of p85α and α-pAkt were decreased, and α-Akt expression was increased (all P<0.05), while the expressions of MMP-9 and E-cadherin showed no significant change (both P>0.05); all the studied parameters in SW1990 cells transfected with negative control siRNA seqeunces had no obvious change (all P>0.05). Co-immunoprecipitation assay showed that there was interaction between AFAP-1L2 and p85α protein in SW1990 cells. Conclusion: AFAP-1L2 may regulate PI3K/Akt pathway through interaction with p85α and thereby influence the migration and invasion of pancreatic cancer cells, and down-regulating AFAP-1L2 expression can decrease the migration and invasion ability of pancreatic cancer cells.