TGF-beta 1 stimulation of cell locomotion utilizes the hyaluronan receptor RHAMM and hyaluronan.
TGF-beta is a potent stimulator of motility in a variety of cell types. It has recently been shown that hyaluronan (HA) can directly promote locomotion of cells through interaction with the HA receptor RHAMM. We have investigated the role of RHAMM and HA in TGF-beta-stimulated locomotion and show that TGF-beta triggers the transcription, synthesis and membrane expression of the RHAMM receptor and the secretion of HA coincident with the induction of the locomotory response. This was demonstrated by both incubating cells with exogenous TGF-beta 1 and by stimulating the production of bioactive TGF-beta 1 in tumor cells transfected with TGF-beta 1 under the control of the metallothionein promoter. TGF-beta 1-induced locomotion was suppressed by antibodies that prevented HA/RHAMM interaction, using polyclonal antibodies to either RHAMM fusion protein or RHAMM peptides, or mAbs to purified RHAMM. Peptides corresponding to the HA-binding motif of RHAMM also suppressed TGF-beta 1-induced increases in motility rate. Spontaneous locomotion of fibrosarcoma cells was blocked by neutralizing secreted TGF-beta with panspecific TGF-beta antibodies and by inhibition of TGF-beta 1 secretion with antisense oligonucleotides. Polyclonal anti-RHAMM fusion protein antibodies and peptide from the RHAMM HA-binding motif also suppressed the spontaneous motility rate of fibrosarcoma cells. These data suggest that fibrosarcoma cell locomotion requires TGF-beta, and the pathway by which TGF-beta stimulates locomotion uses the HA receptor RHAMM and HA.
Samuel
S K
SK
Manitoba Institute of Cell Biology, University of Manitoba, Winnipeg, Canada.
Hurta
R A
RA
Spearman
M A
MA
Wright
J A
JA
Turley
E A
EA
Greenberg
A H
AH
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects*; D005354:Fibrosarcoma; D011905:Genes, ras; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid / Q000096:biosynthesis / Q000502:physiology*; D019868:Kanamycin Kinase; D007700:Kinetics; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D009842:Oligopeptides / Q000494:pharmacology; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000096:biosynthesis; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000187:drug effects / Q000502:physiology*; D016201:Receptors, Lymphocyte Homing / Q000096:biosynthesis / Q000187:drug effects / Q000502:physiology*; D013997:Time Factors; D014158:Transcription, Genetic; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000096:biosynthesis* / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D009838:Oligodeoxyribonucleotides; D016376:Oligonucleotides, Antisense; D009842:Oligopeptides; D011956:Receptors, Cell Surface; D016201:Receptors, Lymphocyte Homing; D016212:Transforming Growth Factor beta; D006820:Hyaluronic Acid; D017853:Phosphotransferases (Alcohol Group Acceptor); D019868:Kanamycin Kinase