Motility analysis of pancreatic adenocarcinoma cells reveals a role for the atypical zeta isoform of protein kinase C in cancer cell movement.
The acquisition of an invasive and metastatic phenotype is accompanied by profound alterations of intracellular mechanisms controlling cell movement. Analysis of quantitative parameters of cell motility in cancer cells may help in the identification of intracellular signaling events determining invasion and metastasis. Here we developed a novel procedure of quantification of cell motility based on time-lapse video microscopy and digital image analysis. Three kinetic parameters, including area change, plasma membrane remodeling, and speed of linear movement, are quantified and combined in one single, time-normalized value we defined motility score (MS). Through calculation of the MS for various human pancreatic adenocarcinoma cell subclones, we identified clones characterized by low or high spontaneous motility in vitro. Analysis of the signaling mechanisms involved in the regulation of pancreatic adenocarcinoma cell motility showed that the atypical zeta isozyme of the serine-threonine protein kinase C (PKC) plays a critical role in maintaining a high MS in motile subclones, as demonstrated by the inhibitory effect of cell permeable peptides with sequence corresponding to the pseudosubstrate inhibitory region of the atypical zeta PKC. Other PKC isozymes, either classic or novel, seem not involved. Furthermore, biochemical analysis showed that in motile cells, zeta PKC is constitutively associated with the plasma membrane, whereas in nonmotile cells, zeta PKC is totally excluded from the plasma membrane. These data suggest that the disregulation of the function of atypical zeta PKC might be involved in the acquisition of an invasive and metastatic phenotype in pancreatic adenocarcinoma cells.
Laudanna
Carlo
C
Department of Pathology, University of Verona, Italy. carlo.laudanna@univr.it
Sorio
Claudio
C
Tecchio
Cristina
C
Butcher
Eugene C
EC
Bonora
Antonio
A
Bassi
Claudio
C
Scarpa
Aldo
A
D016475:3T3 Cells; D000230:Adenocarcinoma / Q000201:enzymology* / Q000473:pathology / Q000503:physiopathology; D000818:Animals; D002465:Cell Movement / Q000502:physiology; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007527:Isoenzymes / Q000378:metabolism*; D051379:Mice; D018715:Microscopy, Video; D010190:Pancreatic Neoplasms / Q000201:enzymology* / Q000473:pathology / Q000503:physiopathology; D011493:Protein Kinase C / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D000099221:Protein Kinase C zeta
D007527:Isoenzymes; D011493:Protein Kinase C; D000099221:Protein Kinase C zeta