Our understanding of the mechanism regulating pancreatic cancer metastatic phenotype is limited. We analyzed the role of RHOA and PRKCZ in the motility attitude of two subclones of SUIT−2 (S2), a cell line derived from metastatic pancreatic adenocarcinoma (PDAC), using RHOA and PRKCZ cell−permeable inhibitory peptides.

Adhesion assays, cell permeable peptides, RHOA activity assay, western blotting.

When used in combination cell−permeable inhibitory peptides partially inhibited cell adhesion by about 50% in subclone S2−CP9. In subclone S2m, the effect was limited to 15% inhibition. In a wound healing assay, S2−CP9 was sensitive only to treatment with the combination of both RHOA and PRKCZ inhibitory peptides. Conversely, S2m was unable to migrate toward both ends of the wound in basal conditions, even if its motility score overlapped that of S2−CP9. Migration of cells through a membrane with 8 um pores was completely abolished in both clones by individual treatment with RHOA and PRKCZ inhibitory peptides.

Herein, we demonstrate a critical role for RHOA and PRKCZ in the regulation of different aspects of cell motility of PDAC and demonstrate the need to inhibit both pathways to obtain a functionally relevant effect in most assays. These results indicate that RHOA and PRKCZ, and their downstream effectors, can represent important pharmacological targets that could potentially control the highly metastatic attitude of PDAC.

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