Changes in the expression levels of alpha1,3 FucT-VII and carbohydrate product SLe X have been reported in certain types of malignant transformations. However, the specific role of FucT-VII in human colorectal carcinoma is still not clear. We evaluated the expression of FucT-VII in tumor specimens from 30 colorectal carcinoma patients by RT-PCR and immunohistochemistry. The alteration of metastatic potential of LOVO cells before and after alpha1,3 FucT-VII gene transfection was also assayed. The expression change of glycoprotein CD24 and carbohydrate antigen SLe X after stable transfection of LOVO was also examined in vitro. Higher mRNA and protein expression of FucT-VII were observed in colorectal tumors compared with adjacent normal ones. The cell chemotactic migration and invasion were enhanced after alpha1,3 FucT-VII transfection in LOVO cells. There was a positive correlation between alpha1,3 FucT-VII mRNA expression and lymph node status (p=0.009) and AJCC stage (p=0.007), similar correlation was also observed at protein level (p=0.042 and 0.022, respectively). Overexpression of alpha1,3 FucT-VII promoted the expression of CD24 and SLe X in LOVO cells. Our findings suggest that alpha1,3 FucT-VII might play a role in colorectal carcinoma metastases by promoting the carbohydration of glycoprotein CD24.
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J Biochem
November 2004
Pharmacology Research Laboratories, Tanabe Seiyaku Co., Ltd., 16-89 Kashima 3-chome, Yodogawa-ku, Osaka 532-8505, Japan.
The biosynthesis of the carbohydrate antigen sialyl Lewis X (sLe(x)) in human leukocytes is mediated by alpha1-3 fucosyltransferase-VII (FucT-VII), which catalyzes the transfer of fucose from GDP-beta-fucose to the 3-OH of alpha2-3 sialyl N-acetyllactosamine (SA-LN). We developed a simple method for quantitating the reaction product of FucT-VII involving Anion-Exchange Chromatography (AEC). The AEC assay involved the separation of a radio-labeled acceptor from the unreacted nucleotide sugars with 0-0.
View Article and Find Full Text PDFBiochem Biophys Res Commun
November 2004
Pharmacology Research Laboratories, Tanabe Seiyaku Co., Ltd., 16-89 Kashima 3-chome, Yodogawa-ku, Osaka 532-8505, Japan.
The sialyl Lewis X (sLe(x)) determinant on leukocytes serves as a ligand for selectin family cell adhesion molecules, and selectin-carbohydrate interaction is considered to play an important role in the process of leukocyte extravasation during inflammation. Among several alpha1-3 fucosyltransferases (FucTs), FucT-VII plays a critical role in the biosynthesis of sLe(x)-epitopes. Therefore, small molecules specifically designed to inhibit the FucT-VII enzyme may have potential as anti-inflammatory agents.
View Article and Find Full Text PDFJ Cell Biol
July 1998
Department of Microbiology/Immunology, Northwestern University Medical School, Chicago, Illinois 60611, USA.
Interactions between P-selectin, expressed on endothelial cells and activated platelets, and its leukocyte ligand, a homodimer termed P-selectin glycoprotein ligand-1 (PSGL-1), mediate the earliest adhesive events during an inflammatory response. To investigate whether dimerization of PSGL-1 is essential for functional interactions with P-selectin, a mutant form of PSGL-1 was generated in which the conserved membrane proximal cysteine was mutated to alanine (designated C320A). Western blotting under both denaturing and native conditions of the C320A PSGL-1 mutant isolated from stably transfected cells revealed expression of only a monomeric form of PSGL-1.
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