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Previous work demonstrated that alpha-thrombin promoted tumor cell adhesion to endothelium and extracellular matrix as well as enhanced the metastatic capacity of tumor cells. This study was initiated to investigate whether the thrombin effect on tumor cells is mediated through the "tethered ligand" thrombin receptor. RT-PCR analysis using primers based on the human thrombin receptors detected mRNA in human colon adenocarcinoma cells (clone A), whose authenticity was confirmed by Southern hybridization. The presence of thrombin receptor mRNA in rat (W256 carcinosarcoma) and mouse (melanoma) tumor cells was demonstrated by RT-PCR/Southern blotting using species-specific PCR primers. Sequencing of the PCR fragment of clone A cells revealed complete homology with the reported human cDNA sequence. Subsequently, tumor cells derived from three species, i.e., human, rat, and mouse, were found to express the thrombin receptor protein as revealed by immunoblotting using ligand peptide-derived mAb ATAP138, whose reactivity towards the M(r) approximately 66,000, potential thrombin receptor was blocked by preincubating the antibody with the immunogen peptide SFLLRNPNDKYEPF (TRP 14). Finally, peptides TRP 14 and TRP 7 (SFLLRNP), but not TRP 5 (FLLRN), were found to mimic alpha-thrombin in stimulating tumor cell adhesion to fibronectin, suggesting that the thrombin receptors expressed on solid tumor cells are biologically functional.
M Z Wojtukiewicz, and
D G Tang, and
E Ben-Josef, and
C Renaud, and
D A Walz, and
K V Honn
July 1996,
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M Z Wojtukiewicz, and
D G Tang, and
E Ben-Josef, and
C Renaud, and
D A Walz, and
K V Honn
January 1997,
Blood coagulation & fibrinolysis : an international journal in haemostasis and thrombosis,
M Z Wojtukiewicz, and
D G Tang, and
E Ben-Josef, and
C Renaud, and
D A Walz, and
K V Honn
February 1997,
Molecular pharmacology,
M Z Wojtukiewicz, and
D G Tang, and
E Ben-Josef, and
C Renaud, and
D A Walz, and
K V Honn
March 1996,
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M Z Wojtukiewicz, and
D G Tang, and
E Ben-Josef, and
C Renaud, and
D A Walz, and
K V Honn
September 1994,
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M Z Wojtukiewicz, and
D G Tang, and
E Ben-Josef, and
C Renaud, and
D A Walz, and
K V Honn
November 1993,
The American journal of physiology,
M Z Wojtukiewicz, and
D G Tang, and
E Ben-Josef, and
C Renaud, and
D A Walz, and
K V Honn
August 1994,
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M Z Wojtukiewicz, and
D G Tang, and
E Ben-Josef, and
C Renaud, and
D A Walz, and
K V Honn
March 1996,
European journal of immunology,
M Z Wojtukiewicz, and
D G Tang, and
E Ben-Josef, and
C Renaud, and
D A Walz, and
K V Honn
February 1992,
Biochemical and biophysical research communications,
M Z Wojtukiewicz, and
D G Tang, and
E Ben-Josef, and
C Renaud, and
D A Walz, and
K V Honn
August 1998,
Journal of biochemistry,
