Specific sequences of fibronectin activate the protein kinase C signal transduction pathway in invasive bladder cancer.
The mechanism of human bladder cancer cell invasion is not clear, but it appears that extracellular matrix components, such as fibronectin, may be involved. To investigate the role of fibronectin in tumor cell invasion and progression, we used an in vitro invasion assay to define the motility stimulating fragment of fibronectin for invasive human bladder cancer T24 cells. Using a modified Boyden chamber assay and purified fragments of fibronectin, we demonstrated that both the 120 kDa chymotrypsin generated fragment of fibronectin (containing the cell attachment RGD motif and additional sequences towards the carboxyl-terminal heparin binding domain), as well as the trypsin generated 60 kDa fragment of fibronectin (containing the carboxyl-terminal heparin binding domain and additional sequences towards the cell attachment RGD motif), were able to stimulate the migration of invasive human bladder cancer T24 cells. Control fragments containing only the amino-terminal gelatin binding region of fibronectin did not stimulate the motility of the human bladder cancer T24 cells. To determine the molecular mechanism in which these fragments may stimulate the migration of the T24 cells, we assayed for intracellular signal transduction pathway protein kinase C (PKC). We demonstrated that both the 120 kDa and the 60 kDa fragments were able to stimulate the activation of protein kinase C. Non-motility stimulating fragments of fibronectin were not able to activate protein kinase C. We conclude that the PKC signal transduction pathway may be involved in matrix mediated motility, and suggest that the inhibition of such pathway(s) may alter the malignant phenotype of human bladder cancer.
Margolis
E J
EJ
Department of Urology, Mount Sinai School of Medicine, New York, 10029, USA.
Choi
J C
JC
Shu
W P
WP
Liu
B C
BC
D000595:Amino Acid Sequence; D001665:Binding Sites; D002465:Cell Movement / Q000502:physiology; D002633:Chemotaxis / Q000502:physiology; D004789:Enzyme Activation; D005353:Fibronectins / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000502:physiology; D011493:Protein Kinase C / Q000378:metabolism / Q000502:physiology*; D012680:Sensitivity and Specificity; D015398:Signal Transduction / Q000502:physiology*; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms / Q000201:enzymology / Q000473:pathology* / Q000503:physiopathology*
D005353:Fibronectins; D009842:Oligopeptides; C047981:arginyl-glycyl-aspartic acid; D011493:Protein Kinase C