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The pathogenesis of pancreatic carcinoma is driven by the tumor cells ability to migrate causing invasion and metastases. The correlation between the aberrant expression of basement membrane proteins and the process of tumor invasion and metastasis has not been fully determined.In the present study, the influence of laminin, fibronectin, and collagen type IV on migratory activity of 5 different cell lines has been investigated at the level of a single tumor cell using 3-dimensional time-lapse microscopy.All investigated cell lines have shown a high baseline migration that varied between 6.2 ± 3.6 and 20.6 ± 6.8 μm/h. The addition of laminin, fibronectin, and collagen type IV to collagen type I matrix has significantly increased tumor cell migration. Tumor cell migration was strongly inhibited after treating the tumor cells with anti-β1 monoclonal antibodies. An abundant and continuous expression of laminin, fibronectin, and collagen type IV was found on the basement membrane of perineurium, which sharply promoted tumor cell invasion.The continuous presentation of the basement membrane proteins by perineurium contributes to the affinity of pancreatic cancer cells for the perineural tumor invasion. Blockade of integrins could represent a possible approach to control the basement membrane-guided tumor spread.