BackgroundPancreatic ductal adenocarcinoma (PDAC) is one of the deadliest cancers based on five-year survival rates. Genes contributing to chemoresistance represent novel therapeutic targets that can improve treatment response. Increased expression ofANGPTL4in tumors correlates with poor outcomes in pancreatic cancer.MethodsWe used statistical analysis of publicly available gene expression data (TCGA-PAAD) to test whether expression ofANGPTL4and its downstream targets,ITGB4andAPOL1, were correlated with patient survival. We measured the impact ofANGPTL4overexpression in a common pancreatic cancer cell line, MIA PaCa-2 cells, using CRISPRa for overexpression and DsiRNA for knockdown. We characterized global gene expression changes associated with high levels ofANGPTL4and response to gemcitabine treatment using RNA-sequencing. Gemcitabine dose response curves were calculated on modified cell lines by measuring cell viability with CellTiter-Glo (Promega). Impacts on cell migration were measured using a time course scratch assay.ResultsWe show thatANGPTL4overexpression leads to in vitro resistance to gemcitabine and reduced survival times in patients. Overexpression ofANGPTL4induces transcriptional signatures of tumor invasion and metastasis, proliferation and differentiation, and inhibition of apoptosis. Analyses revealed an overlapping signature of genes associated with bothANGPTL4activation and gemcitabine response. Increased expression of the genes in this signature in patient PDAC tissues was significantly associated with shorter patient survival. We identified 42 genes that were both co-regulated withANGPTL4and were responsive to gemcitabine treatment.ITGB4andAPOL1were among these genes. Knockdown of either of these genes in cell lines overexpressingANGPTL4reversed the observed gemcitabine resistance and inhibited cellular migration associated with epithelial to mesenchymal transition (EMT) andANGPTL4overexpression.ConclusionsThese data suggest thatANGPTL4promotes EMT and regulates the genesAPOL1andITGB4. Importantly, we show that inhibition of both targets reverses chemoresistance and decreases migratory potential. Our findings have revealed a novel pathway regulating tumor response to treatment and suggest relevant therapeutic targets in pancreatic cancer.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12885-023-11010-1.