RHOA and PRKCZ control different aspects of cell motility in pancreatic cancer metastatic clones.
BACKGROUND
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 the pancreatic adenocarcinoma cell line SUIT-2 (S2), with different in vivo metastatic potential in nude mice: S2-m with a low metastatic potential and highly metastatic S2-CP9 using RHOA and PRKCZ cell-permeable inhibitory peptides.


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


RESULTS
When used in combination cell-permeable inhibitory peptides partially inhibited cell adhesion by about 50% in clone S2-CP9. In clone S2-m, 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, S2-m was unable to migrate toward both ends of the wound in basal conditions. Migration of cells through a membrane with 8 mum pores was completely abolished in both clones by individual treatment with RHOA and PRKCZ inhibitory peptides.


CONCLUSIONS
Herein, we demonstrate a critical role for RHOA and PRKCZ in the regulation of different aspects of cell motility of pancreatic adenocarcinoma 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.
Department of Pathology, University of Verona, Strada Le Grazie 8, Verona, Italy.
Marco
M
Della Peruta
Cinzia
C
Giagulli
Carlo
C
Laudanna
Aldo
A
Scarpa
Claudio
C
Sorio
D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002999:Clone Cells; D008175:Lung Neoplasms / Q000201:enzymology* / Q000556:secondary*; D051379:Mice; D008819:Mice, Nude; D010190:Pancreatic Neoplasms / Q000201:enzymology* / Q000473:pathology*; D010455:Peptides / Q000494:pharmacology; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism*; D014945:Wound Healing / Q000187:drug effects; D020742:rhoA GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*; D000099221:Protein Kinase C zeta
D010455:Peptides; D011493:Protein Kinase C; D020742:rhoA GTP-Binding Protein; D000099221:Protein Kinase C zeta